研究活動情報

• 2021年03月 日本薬理学会, 学術奨励賞


• 2015年02月 公益財団法人井上科学振興財団, 井上研究奨励賞


• 2014年07月 日本骨代謝学会, 研究奨励賞


• 2014年04月 日本リウマチ学会, 奨励賞

• Sublingual immune cell clusters and dendritic cell distribution in the oral cavity

  Yutaka Kusumoto, Mizuki Ueda, Mayuko Hashimoto, Haruka Takeuchi, Naoko Okada, Junya Yamamoto, Akiko Nishii, Atsuki Fujino, Akiho Kurahashi, Momoka Satoh, Yuki Iwasa, Koki Okamura, Karin Obazaki, Ryoto Kumagai, Naruya Sakamoto, Yuto Tanaka, Yukika Kamiya, Tetsushi Hoshida, Tsuneyasu Kaisho, Hiroaki Hemmi, Tomoya Katakai, Tetsuya Honda, Junichi Kikuta, Kosuke Kataoka, Ryoyo Ikebuchi, Taiki Moriya, Takahiro Adachi, Takeshi Watanabe, Masaru Ishii, Atsushi Miyawaki, Kenji Kabashima, Tatyana Chtanova, Michio Tomura

  American Society for Clinical Investigation, 2024年10月, JCI Insight, 9(21) (21)

  研究論文（学術雑誌）


• Periportal macrophages protect against commensal-driven liver inflammation

  Yu Miyamoto, Junichi Kikuta, Takahiro Matsui, Tetsuo Hasegawa, Kentaro Fujii, Daisuke Okuzaki, Yu-chen Liu, Takuya Yoshioka, Shigeto Seno, Daisuke Motooka, Yutaka Uchida, Erika Yamashita, Shogo Kobayashi, Hidetoshi Eguchi, Eiichi Morii, Karl Tryggvason, Takashi Shichita, Hisako Kayama, Koji Atarashi, Jun Kunisawa, Kenya Honda, Kiyoshi Takeda, Masaru Ishii

  Springer Science and Business Media LLC, 2024年04月, Nature

  研究論文（学術雑誌）


• Protocol for live imaging of transferred mouse bone marrow cells by two-photon microscopy.

  Takao Sudo, Erika Yamashita, Junichi Kikuta, Masaru Ishii

  The in situ behavior of living cells can be visualized by two-photon microscopy. Here, we present a protocol for the live imaging of transferred mouse bone marrow cells by two-photon microscopy. We describe steps for staining and injecting target cells into mice, fixing the skull bone to a head holder and stage, and 4D imaging bone marrow using multi-photon microscopy. We then detail procedures for creating images and analyzing cells. For complete details on the use and execution of this protocol, please refer to Sudo et al. (2021).1.

  2023年12月, STAR protocols, 4(4) (4), 102654 - 102654, 英語, 国際誌

  研究論文（学術雑誌）


• Activation of ILC2s through constitutive IFNγ signaling reduction leads to spontaneous pulmonary fibrosis.

  Natsuko Otaki, Yasutaka Motomura, Tommy Terooatea, S Thomas Kelly, Miho Mochizuki, Natsuki Takeno, Shigeo Koyasu, Miu Tamamitsu, Fuminori Sugihara, Junichi Kikuta, Hideya Kitamura, Yoshiki Shiraishi, Jun Miyanohara, Yuji Nagano, Yuji Saita, Takashi Ogura, Koichiro Asano, Aki Minoda, Kazuyo Moro

  Pulmonary fibrosis (PF), a condition characterized by inflammation and collagen deposition in the alveolar interstitium, causes dyspnea and fatal outcomes. Although the bleomycin-induced PF mouse model has improved our understanding of exogenous factor-induced fibrosis, the mechanism governing endogenous factor-induced fibrosis remains unknown. Here, we find that Ifngr1-/-Rag2-/- mice, which lack the critical suppression factor for group 2 innate lymphoid cells (ILC2), develop PF spontaneously. The onset phase of fibrosis includes ILC2 subpopulations with a high Il1rl1 (IL-33 receptor) expression, and fibrosis does not develop in ILC-deficient or IL-33-deficient mice. Although ILC2s are normally localized near bronchioles and blood vessels, ILC2s are increased in fibrotic areas along with IL-33 positive fibroblasts during fibrosis. Co-culture analysis shows that activated-ILC2s directly induce collagen production from fibroblasts. Furthermore, increased IL1RL1 and decreased IFNGR1 expressions are confirmed in ILC2s from individuals with idiopathic PF, highlighting the applicability of Ifngr1-/-Rag2-/- mice as a mouse model for fibrosis research.

  2023年12月, Nature communications, 14(1) (1), 8120 - 8120, 英語, 国際誌

  研究論文（学術雑誌）


• Analysis of distribution, collection, and confirmation of capacity dependency of small extracellular vesicles toward a therapy for liver cirrhosis.

  Nobutaka Takeda, Atsunori Tsuchiya, Masaki Mito, Kazuki Natsui, Yui Natusi, Yohei Koseki, Kei Tomiyoshi, Fusako Yamazaki, Yuki Yoshida, Hiroyuki Abe, Masayuki Sano, Taketomo Kido, Yusuke Yoshioka, Junichi Kikuta, Tohru Itoh, Ken Nishimura, Masaru Ishii, Takahiro Ochiya, Atsushi Miyajima, Shuji Terai

  BACKGROUND: The progression of liver fibrosis leads to portal hypertension and liver dysfunction. However, no antifibrotic agents have been approved for cirrhosis to date, making them an unmet medical need. Small extracellular vesicles (sEVs) of mesenchymal stem cells (MSCs) are among these candidate agents. In this study, we investigated the effects of sEVs of MSCs, analyzed their distribution in the liver post-administration, whether their effect was dose-dependent, and whether it was possible to collect a large number of sEVs. METHODS: sEVs expressing tdTomato were generated, and their uptake into constituent liver cells was observed in vitro, as well as their sites of uptake and cells in the liver using a mouse model of liver cirrhosis. The efficiency of sEV collection using tangential flow filtration (TFF) and changes in the therapeutic effects of sEVs in a volume-dependent manner were examined. RESULTS: The sEVs of MSCs accumulated mostly in macrophages in damaged areas of the liver. In addition, the therapeutic effect of sEVs was not necessarily dose-dependent, and it reached a plateau when the dosage exceeded a certain level. Furthermore, although ultracentrifugation was commonly used to collect sEVs for research purposes, we verified that TFF could be used for efficient sEV collection and that their effectiveness is not reduced. CONCLUSION: In this study, we identified some unknown aspects regarding the dynamics, collection, and capacity dependence of sEVs. Our results provide important fundamentals for the development of therapies using sEVs and hold potential implications for the therapeutic applications of sEV-based therapies for liver cirrhosis.

  2023年10月, Inflammation and regeneration, 43(1) (1), 48 - 48, 英語, 国際誌

  研究論文（学術雑誌）


• Hematopoietic cell-derived IL-15 supports NK cell development in scattered and clustered localization within the bone marrow.

  Shinya Abe, Takuma Asahi, Takahiro Hara, Guangwei Cui, Akihiro Shimba, Shizue Tani-Ichi, Kohei Yamada, Kazuko Miyazaki, Hitoshi Miyachi, Satsuki Kitano, Naotoshi Nakamura, Junichi Kikuta, Alexis Vandenbon, Masaki Miyazaki, Ryo Yamada, Toshiaki Ohteki, Masaru Ishii, Veronika Sexl, Takashi Nagasawa, Koichi Ikuta

  Natural killer (NK) cells are innate immune cells critical for protective immune responses against infection and cancer. Although NK cells differentiate in the bone marrow (BM) in an interleukin-15 (IL-15)-dependent manner, the cellular source of IL-15 remains elusive. Using NK cell reporter mice, we show that NK cells are localized in the BM in scattered and clustered manners. NK cell clusters overlap with monocyte and dendritic cell accumulations, whereas scattered NK cells require CXCR4 signaling. Using cell-specific IL-15-deficient mice, we show that hematopoietic cells, but not stromal cells, support NK cell development in the BM through IL-15. In particular, IL-15 produced by monocytes and dendritic cells appears to contribute to NK cell development. These results demonstrate that hematopoietic cells are the IL-15 niche for NK cell development in the BM and that BM NK cells are present in scattered and clustered compartments by different mechanisms, suggesting their distinct functions in the immune response.

  2023年09月, Cell reports, 113127 - 113127, 英語, 国際誌

  研究論文（学術雑誌）


• Single-cell transcriptomics of human cholesteatoma identifies an activin A-producing osteoclastogenic fibroblast subset inducing bone destruction

  Kotaro Shimizu, Junichi Kikuta, Yumi Ohta, Yutaka Uchida, Yu Miyamoto, Akito Morimoto, Shinya Yari, Takashi Sato, Takefumi Kamakura, Kazuo Oshima, Ryusuke Imai, Yu-Chen Liu, Daisuke Okuzaki, Tetsuya Hara, Daisuke Motooka, Noriaki Emoto, Hidenori Inohara, Masaru Ishii

  Abstract Cholesteatoma, which potentially results from tympanic membrane retraction, is characterized by intractable local bone erosion and subsequent hearing loss and brain abscess formation. However, the pathophysiological mechanisms underlying bone destruction remain elusive. Here, we performed a single-cell RNA sequencing analysis on human cholesteatoma samples and identify a pathogenic fibroblast subset characterized by abundant expression of inhibin βA. We demonstrate that activin A, a homodimer of inhibin βA, promotes osteoclast differentiation. Furthermore, the deletion of inhibin βA /activin A in these fibroblasts results in decreased osteoclast differentiation in a murine model of cholesteatoma. Moreover, follistatin, an antagonist of activin A, reduces osteoclastogenesis and resultant bone erosion in cholesteatoma. Collectively, these findings indicate that unique activin A-producing fibroblasts present in human cholesteatoma tissues are accountable for bone destruction via the induction of local osteoclastogenesis, suggesting a potential therapeutic target.

  Springer Science and Business Media LLC, 2023年08月, Nature Communications, 14(1) (1)

  研究論文（学術雑誌）


• JAK inhibition ameliorates bone destruction by simultaneously targeting mature osteoclasts and their precursors.

  Shinya Yari, Junichi Kikuta, Hotaka Shigyo, Yu Miyamoto, Daisuke Okuzaki, Yuki Furusawa, Masafumi Minoshima, Kazuya Kikuchi, Masaru Ishii

  BACKGROUND: Rheumatoid arthritis (RA) is characterized by chronic inflammation and resultant cartilage/bone destruction because of aberrantly activated osteoclasts. Recently, novel treatments with several Janus kinase (JAK) inhibitors have been shown to successfully ameliorate arthritis-related inflammation and bone erosion, although their mechanisms of action for limiting bone destruction remain unclear. Here, we examined the effects of a JAK inhibitor on mature osteoclasts and their precursors by intravital multiphoton imaging. METHODS: Inflammatory bone destruction was induced by local injection of lipopolysaccharides into transgenic mice carrying reporters for mature osteoclasts or their precursors. Mice were treated with the JAK inhibitor, ABT-317, which selectively inhibits the activation of JAK1, and then subjected to intravital imaging with multiphoton microscopy. We also used RNA sequencing (RNA-Seq) analysis to investigate the molecular mechanism underlying the effects of the JAK inhibitor on osteoclasts. RESULTS: The JAK inhibitor, ABT-317, suppressed bone resorption by blocking the function of mature osteoclasts and by targeting the migratory behaviors of osteoclast precursors to the bone surface. Further exhaustive RNA-Seq analysis demonstrated that Ccr1 expression on osteoclast precursors was suppressed in the JAK inhibitor-treated mice; the CCR1 antagonist, J-113863, altered the migratory behaviors of osteoclast precursors, which led to the inhibition of bone destruction under inflammatory conditions. CONCLUSIONS: This is the first study to determine the pharmacological actions by which a JAK inhibitor blocks bone destruction under inflammatory conditions; this inhibition is beneficial because of its dual effects on both mature osteoclasts and immature osteoclast precursors.

  2023年03月, Inflammation and regeneration, 43(1) (1), 18 - 18, 英語, 国際誌

  研究論文（学術雑誌）


• Progressive differentiation toward the long-lived plasma cell compartment in the bone marrow

  Takuya Koike, Kentaro Fujii, Kohei Kometani, Noah S. Butler, Kenji Funakoshi, Shinya Yari, Junichi Kikuta, Masaru Ishii, Tomohiro Kurosaki, Wataru Ise

  The longevity of plasma cells is dependent on their ability to access and reside in so-called niches that are predominantly located in the bone marrow. Here, by employing a traceable method to label recently generated plasma cells, we showed that homeostatic plasma cells in the bone marrow and spleen were continuously replenished by newly generated B220hiMHC-IIhi populations that progressively differentiated into B220loMHC-IIlo long-lived plasma cell (LLPC) populations. We also found that, in the bone marrow, germinal center (GC)–independent and GC-dependent plasma cells decayed similarly upon NP-CGG engagement, and both entered the B220loMHC-IIlo LLPC pool. Compared with NP+B220hiMHC-IIhi plasma cells, NP+B220loMHC-IIlo cells were more immobilized in the bone marrow niches and showed better survival potential. Thus, our results suggest that the adhesion status of bone marrow plasma cells is dynamically altered during their differentiation and is associated with provision of survival signals.

  Rockefeller University Press, 2023年02月, Journal of Experimental Medicine, 220(2) (2)

  研究論文（学術雑誌）


• In vivo induction of activin A-producing alveolar macrophages supports the progression of lung cell carcinoma

  Seiji Taniguchi, Takahiro Matsui, Kenji Kimura, Soichiro Funaki, Yu Miyamoto, Yutaka Uchida, Takao Sudo, Junichi Kikuta, Tetsuya Hara, Daisuke Motooka, Yu-Chen Liu, Daisuke Okuzaki, Eiichi Morii, Noriaki Emoto, Yasushi Shintani, Masaru Ishii

  Abstract Alveolar macrophages (AMs) are crucial for maintaining normal lung function. They are abundant in lung cancer tissues, but their pathophysiological significance remains unknown. Here we show, using an orthotopic murine lung cancer model and human carcinoma samples, that AMs support cancer cell proliferation and thus contribute to unfavourable outcome. Inhibin beta A (INHBA) expression is upregulated in AMs under tumor-bearing conditions, leading to the secretion of activin A, a homodimer of INHBA. Accordingly, follistatin, an antagonist of activin A is able to inhibit lung cancer cell proliferation. Single-cell RNA sequence analysis identifies a characteristic subset of AMs specifically induced in the tumor environment that are abundant in INHBA, and distinct from INHBA-expressing AMs in normal lungs. Moreover, postnatal deletion of INHBA/activin A could limit tumor growth in experimental models. Collectively, our findings demonstrate the critical pathological role of activin A-producing AMs in tumorigenesis, and provides means to clearly distinguish them from their healthy counterparts.

  Springer Science and Business Media LLC, 2023年01月, Nature Communications, 14(1) (1)

  研究論文（学術雑誌）


• Label‐free multiphoton excitation imaging as a promising diagnostic tool for breast cancer

  Takahiro Matsui, Akio Iwasa, Masafumi Mimura, Seiji Taniguchi, Takao Sudo, Yutaka Uchida, Junichi Kikuta, Hidetomo Morizono, Rie Horii, Yuichi Motoyama, Eiichi Morii, Shinji Ohno, Yasujiro Kiyota, Masaru Ishii

  Wiley, 2022年08月, Cancer Science, 113(8) (8), 2916 - 2925

  研究論文（学術雑誌）


• A spontaneous pulmonary fibrosis mouse model identifies a mechanism for ILC2-mediated irreversible fibrosis

  Kazuyo Moro, Natsuko Otaki, Yasutaka Motomura, Tommy Terooatea, S. Thomas Kelly, Miho Mochizuki, Natsuki Takeno, Shigeo Koyasu, Fuminori Sugihara, Junichi Kikuta, Hideya Kitamura, Yoshiki Shiraishi, Jun Miyanohara, Yuji Nagano, Yuji Saita, Takashi Ogura, Koichiro Asano, Aki Minoda

  Abstract Pulmonary fibrosis (PF) is characterised by inflammation and collagen deposition in the alveolar interstitium, leading to dyspnoea and death. However, the comprehensive pathogenesis of PF remains unclear due to the lack of spontaneous fibrosis mouse models. Here, we found that Ifngr1^-/-Rag2^-/- mice, lacking the mechanisms to suppress group 2 and 3 innate lymphoid cells (ILC2s and ILC3s), developed severe PF spontaneously. In Ifngr1^-/-Rag2^-/- mice, Il1rl1^hiIl13^hi-ILC2 subpopulation was increased at disease-onset phase before collagen production began. Further, defects in ILCs or IL-33, the strong ILC2 activator, prevented PF development. ILC2s directly induce collagen production by fibroblasts in vitro, and fibroblasts started to produce IL-33 in the chronic phase, presumably forming a positive feedback loop between fibroblasts and ILC2s leading to irreversible fibrosis. Moreover, the increased IL1RL1 and IL13 and decreased IFNGR1 expression levels in ILC2s from human idiopathic PF patients highlight the significance of the novel mouse model in fibrosis research.

  Research Square Platform LLC, 2022年05月


• Determination of the physiological range of oxygen tension in bone marrow monocytes using two-photon phosphorescence lifetime imaging microscopy.

  Ayako Narazaki, Reito Shimizu, Toshitada Yoshihara, Junichi Kikuta, Reiko Sakaguchi, Seiji Tobita, Yasuo Mori, Masaru Ishii, Keizo Nishikawa

  Oxygen is a key regulator of both development and homeostasis. To study the role of oxygen, a variety of in vitro and ex vivo cell and tissue models have been used in biomedical research. However, because of ambiguity surrounding the level of oxygen that cells experience in vivo, the cellular pathway related to oxygenation state and hypoxia have been inadequately studied in many of these models. Here, we devised a method to determine the oxygen tension in bone marrow monocytes using two-photon phosphorescence lifetime imaging microscopy with the cell-penetrating phosphorescent probe, BTPDM1. Phosphorescence lifetime imaging revealed the physiological level of oxygen tension in monocytes to be 5.3% in live mice exposed to normal air. When the mice inhaled hypoxic air, the level of oxygen tension in bone marrow monocytes decreased to 2.4%. By performing in vitro cell culture experiment within the physiological range of oxygen tension, hypoxia changed the molecular phenotype of monocytes, leading to enhanced the expression of CD169 and CD206, which are markers of a unique subset of macrophages in bone marrow, osteal macrophages. This current study enables the determination of the physiological range of oxygen tension in bone marrow with spatial resolution at a cellular level and application of this information on oxygen tension in vivo to in vitro assays. Quantifying oxygen tension in tissues can provide invaluable information on metabolism under physiological and pathophyisological conditions. This method will open new avenues for research on oxygen biology.

  2022年03月, Scientific reports, 12(1) (1), 3497 - 3497, 英語, 国際誌

  研究論文（学術雑誌）


• Osteoblast-derived vesicles induce a switch from bone-formation to bone-resorption in vivo.

  Maki Uenaka, Erika Yamashita, Junichi Kikuta, Akito Morimoto, Tomoka Ao, Hiroki Mizuno, Masayuki Furuya, Tetsuo Hasegawa, Hiroyuki Tsukazaki, Takao Sudo, Keizo Nishikawa, Daisuke Okuzaki, Daisuke Motooka, Nobuyoshi Kosaka, Fuminori Sugihara, Thomas Boettger, Thomas Braun, Takahiro Ochiya, Masaru Ishii

  Bone metabolism is regulated by the cooperative activity between bone-forming osteoblasts and bone-resorbing osteoclasts. However, the mechanisms mediating the switch between the osteoblastic and osteoclastic phases have not been fully elucidated. Here, we identify a specific subset of mature osteoblast-derived extracellular vesicles that inhibit bone formation and enhance osteoclastogenesis. Intravital imaging reveals that mature osteoblasts secrete and capture extracellular vesicles, referred to as small osteoblast vesicles (SOVs). Co-culture experiments demonstrate that SOVs suppress osteoblast differentiation and enhance the expression of receptor activator of NF-κB ligand, thereby inducing osteoclast differentiation. We also elucidate that the SOV-enriched microRNA miR-143 inhibits Runt-related transcription factor 2, a master regulator of osteoblastogenesis, by targeting the mRNA expression of its dimerization partner, core-binding factor β. In summary, we identify SOVs as a mode of cell-to-cell communication, controlling the dynamic transition from bone-forming to bone-resorbing phases in vivo.

  2022年02月, Nature communications, 13(1) (1), 1066 - 1066, 英語, 国際誌

  研究論文（学術雑誌）


• The Effects of Vitamin D on Immune System and Inflammatory Diseases.

  Tomoka Ao, Junichi Kikuta, Masaru Ishii

  Immune cells, including dendritic cells, macrophages, and T and B cells, express the vitamin D receptor and 1α-hydroxylase. In vitro studies have shown that 1,25-dihydroxyvitamin D, the active form of vitamin D, has an anti-inflammatory effect. Recent epidemiological evidence has indicated a significant association between vitamin D deficiency and an increased incidence, or aggravation, of infectious diseases and inflammatory autoimmune diseases, such as rheumatoid arthritis, systemic lupus erythematosus, and multiple sclerosis. However, the impact of vitamin D on treatment and prevention, particularly in infectious diseases such as the 2019 coronavirus disease (COVID-19), remains controversial. Here, we review recent evidence associated with the relationship between vitamin D and inflammatory diseases and describe the underlying immunomodulatory effect of vitamin D.

  2021年11月, Biomolecules, 11(11) (11), 英語, 国際誌

  研究論文（学術雑誌）


• Anti-Siglec-15 antibody suppresses bone resorption by inhibiting osteoclast multinucleation without attenuating bone formation.

  Hiroyuki Tsukazaki, Junichi Kikuta, Tomoka Ao, Akito Morimoto, Chie Fukuda, Eisuke Tsuda, Masafumi Minoshima, Kazuya Kikuchi, Takashi Kaito, Masaru Ishii

  Anti-resorptive drugs are widely used for the treatment of osteoporosis, but excessive inhibition of osteoclastogenesis can suppress bone turnover and cause the deterioration of bone quality. Sialic acid-binding immunoglobulin-like lectin 15 (Siglec-15) is a transmembrane protein expressed on osteoclast precursor cells and mature osteoclasts. Siglec-15 regulates proteins containing immunoreceptor tyrosine-based activation motif (ITAM) domains, which then induce nuclear factor of activated T-cells 1 (NFATc1), a master transcription factor of osteoclast differentiation. Anti-Siglec-15 antibody modulates ITAM signaling in osteoclast precursors and inhibits the maturation of osteoclasts in vitro. However, in situ pharmacological effects, particularly during postmenopausal osteoporosis, remain unclear. Here, we demonstrated that anti-Siglec-15 antibody treatment protected against ovariectomy-induced bone loss by specifically inhibiting the generation of multinucleated osteoclasts in vivo. Moreover, treatment with anti-Siglec-15 antibody maintained bone formation to a greater extent than with risedronate, the first-line treatment for osteoporosis. Intravital imaging revealed that anti-Siglec-15 antibody treatment did not cause a reduction in osteoclast motility, whereas osteoclast motility declined following risedronate treatment. We evaluated osteoclast activity using a pH-sensing probe and found that the bone resorptive ability of osteoclasts was lower following anti-Siglec-15 antibody treatment compared to after risedronate treatment. Our findings suggest that anti-Siglec-15 treatment may have potential as an anti-resorptive therapy for osteoporosis, which substantially inhibits the activity of osteoclasts while maintaining physiological bone coupling.

  2021年11月, Bone, 152, 116095 - 116095, 英語, 国際誌

  研究論文（学術雑誌）


• Osteoclasts adapt to physioxia perturbation through DNA demethylation.

  Keizo Nishikawa, Shigeto Seno, Toshitada Yoshihara, Ayako Narazaki, Yuki Sugiura, Reito Shimizu, Junichi Kikuta, Reiko Sakaguchi, Norio Suzuki, Norihiko Takeda, Hiroaki Semba, Masamichi Yamamoto, Daisuke Okuzaki, Daisuke Motooka, Yasuhiro Kobayashi, Makoto Suematsu, Haruhiko Koseki, Hideo Matsuda, Masayuki Yamamoto, Seiji Tobita, Yasuo Mori, Masaru Ishii

  Oxygen plays an important role in diverse biological processes. However, since quantitation of the partial pressure of cellular oxygen in vivo is challenging, the extent of oxygen perturbation in situ and its cellular response remains underexplored. Using two-photon phosphorescence lifetime imaging microscopy, we determine the physiological range of oxygen tension in osteoclasts of live mice. We find that oxygen tension ranges from 17.4 to 36.4 mmHg, under hypoxic and normoxic conditions, respectively. Physiological normoxia thus corresponds to 5% and hypoxia to 2% oxygen in osteoclasts. Hypoxia in this range severely limits osteoclastogenesis, independent of energy metabolism and hypoxia-inducible factor activity. We observe that hypoxia decreases ten-eleven translocation (TET) activity. Tet2/3 cooperatively induces Prdm1 expression via oxygen-dependent DNA demethylation, which in turn activates NFATc1 required for osteoclastogenesis. Taken together, our results reveal that TET enzymes, acting as functional oxygen sensors, regulate osteoclastogenesis within the physiological range of oxygen tension, thus opening new avenues for research on in vivo response to oxygen perturbation.

  2021年10月, EMBO reports, 22(12) (12), e53035, 英語, 国際誌

  研究論文（学術雑誌）


• Group 2 innate lymphoid cells in bone marrow regulate osteoclastogenesis in a reciprocal manner via RANKL, GM-CSF and IL-13.

  Yoshiki Momiuchi, Yasutaka Motomura, Emiko Suga, Hiroki Mizuno, Junichi Kikuta, Akito Morimoto, Miho Mochizuki, Natsuko Otaki, Masaru Ishii, Kazuyo Moro

  Group 2 innate lymphoid cells (ILC2s) are tissue-resident cells that play different roles in different organs by sensing surrounding environmental factors. Initially, it was thought that ILC2s in bone marrow (BM) are progenitors for systemic ILC2s, which migrate to other organs and acquire effector functions. However, accumulating evidence that ILC2s differentiate in peripheral tissues suggests that BM ILC2s may play a specific role in the BM as a unique effector per se. Here, we demonstrate that BM ILC2s highly express the receptor activator of nuclear factor κB ligand (RANKL), a robust cytokine for osteoclast differentiation and activation, and RANKL expression on ILC2s is upregulated by interleukin (IL)-2, IL-7 and all-trans retinoic acid (ATRA). BM ILC2s co-cultured with BM-derived monocyte/macrophage lineage cells (BMMs) in the presence of IL-7 induce the differentiation of tartrate-resistant acid phosphatase (TRAP)-positive osteoclasts in a RANKL-dependent manner. In contrast, BM ILC2s stimulated with IL-33 downregulate RANKL expression and convert BMMs differentiation into M2 macrophage-like cells rather than osteoclasts by granulocyte macrophage colony-stimulating factor (GM-CSF) and IL-13 production. Intravital imaging using two-photon microscopy revealed that a depletion of ILC2s prominently impaired in vivo osteoclast activity in an IL-7 plus ATRA-induced bone loss mouse model. These results suggest that ILC2s regulate osteoclast activation and contribute to bone homeostasis in both steady state and IL-33-induced inflammation.

  2021年09月, International immunology, 33(11) (11), 573 - 585, 英語, 国際誌

  研究論文（学術雑誌）


• Group 2 innate lymphoid cells support hematopoietic recovery under stress conditions

  Takao Sudo, Yasutaka Motomura, Daisuke Okuzaki, Tetsuo Hasegawa, Takafumi Yokota, Junichi Kikuta, Tomoka Ao, Hiroki Mizuno, Takahiro Matsui, Daisuke Motooka, Ryosuke Yoshizawa, Takashi Nagasawa, Yuzuru Kanakura, Kazuyo Moro, Masaru Ishii

  The cell-cycle status of hematopoietic stem and progenitor cells (HSPCs) becomes activated following chemotherapy-induced stress, promoting bone marrow (BM) regeneration; however, the underlying molecular mechanism remains elusive. Here we show that BM-resident group 2 innate lymphoid cells (ILC2s) support the recovery of HSPCs from 5-fluorouracil (5-FU)–induced stress by secreting granulocyte-macrophage colony-stimulating factor (GM-CSF). Mechanistically, IL-33 released from chemo-sensitive B cell progenitors activates MyD88-mediated secretion of GM-CSF in ILC2, suggesting the existence of a B cell–ILC2 axis for maintaining hematopoietic homeostasis. GM-CSF knockout mice treated with 5-FU showed severe loss of myeloid lineage cells, causing lethality, which was rescued by transferring BM ILC2s from wild-type mice. Further, the adoptive transfer of ILC2s to 5-FU–treated mice accelerates hematopoietic recovery, while the reduction of ILC2s results in the opposite effect. Thus, ILC2s may function by “sensing” the damaged BM spaces and subsequently support hematopoietic recovery under stress conditions.

  Rockefeller University Press, 2021年05月, Journal of Experimental Medicine, 218(5) (5)

  研究論文（学術雑誌）


• SLPI is a critical mediator that controls PTH-induced bone formation.

  Akito Morimoto, Junichi Kikuta, Keizo Nishikawa, Takao Sudo, Maki Uenaka, Masayuki Furuya, Tetsuo Hasegawa, Kunihiko Hashimoto, Hiroyuki Tsukazaki, Shigeto Seno, Akira Nakamura, Daisuke Okuzaki, Fuminori Sugihara, Akinori Ninomiya, Takeshi Yoshimura, Ryoko Takao-Kawabata, Hideo Matsuda, Masaru Ishii

  Osteoclastic bone resorption and osteoblastic bone formation/replenishment are closely coupled in bone metabolism. Anabolic parathyroid hormone (PTH), which is commonly used for treating osteoporosis, shifts the balance from osteoclastic to osteoblastic, although it is unclear how these cells are coordinately regulated by PTH. Here, we identify a serine protease inhibitor, secretory leukocyte protease inhibitor (SLPI), as a critical mediator that is involved in the PTH-mediated shift to the osteoblastic phase. Slpi is highly upregulated in osteoblasts by PTH, while genetic ablation of Slpi severely impairs PTH-induced bone formation. Slpi induction in osteoblasts enhances its differentiation, and increases osteoblast-osteoclast contact, thereby suppressing osteoclastic function. Intravital bone imaging reveals that the PTH-mediated association between osteoblasts and osteoclasts is disrupted in the absence of SLPI. Collectively, these results demonstrate that SLPI regulates the communication between osteoblasts and osteoclasts to promote PTH-induced bone anabolism.

  2021年04月, Nature communications, 12(1) (1), 2136 - 2136, 英語, 国際誌

  研究論文（学術雑誌）


• Small extracellular vesicles derived from interferon-γ pre-conditioned mesenchymal stromal cells effectively treat liver fibrosis.

  Suguru Takeuchi, Atsunori Tsuchiya, Takahiro Iwasawa, Shunsuke Nojiri, Takayuki Watanabe, Masahiro Ogawa, Tomoaki Yoshida, Katsunori Fujiki, Yuta Koui, Taketomo Kido, Yusuke Yoshioka, Mayu Fujita, Junichi Kikuta, Tohru Itoh, Masaaki Takamura, Katsuhiko Shirahige, Masaru Ishii, Takahiro Ochiya, Atsushi Miyajima, Shuji Terai

  Mesenchymal stromal cells (MSCs) are used for ameliorating liver fibrosis and aiding liver regeneration after cirrhosis; Here, we analyzed the therapeutic potential of small extracellular vesicles (sEVs) derived from interferon-γ (IFN-γ) pre-conditioned MSCs (γ-sEVs). γ-sEVs effectively induced anti-inflammatory macrophages with high motility and phagocytic abilities in vitro, while not preventing hepatic stellate cell (HSC; the major source of collagen fiber) activation in vitro. The proteome analysis of MSC-derived sEVs revealed anti-inflammatory macrophage inducible proteins (e.g., annexin-A1, lactotransferrin, and aminopeptidase N) upon IFN-γ stimulation. Furthermore, by enabling CX3CR1+ macrophage accumulation in the damaged area, γ-sEVs ameliorated inflammation and fibrosis in the cirrhosis mouse model more effectively than sEVs. Single cell RNA-Seq analysis revealed diverse effects, such as induction of anti-inflammatory macrophages and regulatory T cells, in the cirrhotic liver after γ-sEV administration. Overall, IFN-γ pre-conditioning altered sEVs resulted in efficient tissue repair indicating a new therapeutic strategy.

  2021年03月, NPJ Regenerative medicine, 6(1) (1), 19 - 19, 英語, 国際誌

  研究論文（学術雑誌）


• Intravital imaging of orthotopic and ectopic bone.

  Kunihiko Hashimoto, Takashi Kaito, Junichi Kikuta, Masaru Ishii

  Bone homeostasis is dynamically regulated by a balance between bone resorption by osteoclasts and bone formation by osteoblasts. Visualizing and evaluating the dynamics of bone cells in vivo remain difficult using conventional technologies, including histomorphometry and imaging analysis. Over the past two decades, multiphoton microscopy, which can penetrate thick specimens, has been utilized in the field of biological imaging. Using this innovative technique, the in vivo dynamic motion of bone metabolism-related cells and their interactions has been revealed. In this review, we summarize previous approaches used for bone imaging and provide an overview of current bone tissue imaging methods using multiphoton excitation microscopy.

  2020年11月, Inflammation and regeneration, 40(1) (1), 26 - 26, 英語, 国際誌

  研究論文（学術雑誌）


• Local sympathetic neurons promote neutrophil egress from the bone marrow at the onset of acute inflammation.

  Tomoka Ao, Junichi Kikuta, Takao Sudo, Yutaka Uchida, Kenta Kobayashi, Masaru Ishii

  The sympathetic nervous system plays critical roles in the differentiation, maturation and recruitment of immune cells under homeostatic conditions, and in responses to environmental stimuli, although its role in the migratory control of immune cells during acute inflammation remains unclear. In this study, using an advanced intravital bone imaging system established in our laboratory, we demonstrated that the sympathetic nervous system locally regulates neutrophil egress from the bone marrow for mobilization to inflammatory foci. We found that sympathetic neurons were located close to blood vessels in the bone marrow cavity; moreover, upon lipopolysaccharide (LPS) administration, local sympathectomy delayed neutrophil egress from the bone marrow and increased the proportion of neutrophils that remained in place. We also showed that vascular endothelial cells produced C-X-C motif chemokine ligand 1 (CXCL1), which is responsible for neutrophil egress out of the bone marrow. Its expression was up-regulated during acute inflammation, and was suppressed by β-adrenergic receptor blockade, which was accompanied with inhibition of neutrophil egress into the systemic circulation. Furthermore, systemic β-adrenergic signaling blockade decreased the recruitment of neutrophils in the lung under conditions of acute systemic inflammation. Taken together, the results of this study first suggested a new regulatory system, wherein local sympathetic nervous activation promoted neutrophil egress by enhancing Cxcl1 expression in bone marrow endothelial cells in a β-adrenergic signaling-dependent manner, contributing to the recruitment of neutrophils at the onset of inflammation in vivo.

  Oxford University Press (OUP), 2020年10月, International immunology, 32(11) (11), 727 - 736, 英語, 国際誌

  [査読有り]

  研究論文（学術雑誌）


• Expression of leukotriene B4 receptor 1 defines functionally distinct DCs that control allergic skin inflammation.

  Tomoaki Koga, Fumiyuki Sasaki, Kazuko Saeki, Soken Tsuchiya, Toshiaki Okuno, Mai Ohba, Takako Ichiki, Satoshi Iwamoto, Hirotsugu Uzawa, Keiko Kitajima, Chikara Meno, Eri Nakamura, Norihiro Tada, Yoshinori Fukui, Junichi Kikuta, Masaru Ishii, Yukihiko Sugimoto, Mitsuyoshi Nakao, Takehiko Yokomizo

  Leukotriene B4 (LTB4) receptor 1 (BLT1) is a chemotactic G protein-coupled receptor expressed by leukocytes, such as granulocytes, macrophages, and activated T cells. Although there is growing evidence that BLT1 plays crucial roles in immune responses, its role in dendritic cells remains largely unknown. Here, we identified novel DC subsets defined by the expression of BLT1, namely, BLT1hi and BLT1lo DCs. We also found that BLT1hi and BLT1lo DCs differentially migrated toward LTB4 and CCL21, a lymph node-homing chemoattractant, respectively. By generating LTB4-producing enzyme LTA4H knockout mice and CD11c promoter-driven Cre recombinase-expressing BLT1 conditional knockout (BLT1 cKO) mice, we showed that the migration of BLT1hi DCs exacerbated allergic contact dermatitis. Comprehensive transcriptome analysis revealed that BLT1hi DCs preferentially induced Th1 differentiation by upregulating IL-12p35 expression, whereas BLT1lo DCs accelerated T cell proliferation by producing IL-2. Collectively, the data reveal an unexpected role for BLT1 as a novel DC subset marker and provide novel insights into the role of the LTB4-BLT1 axis in the spatiotemporal regulation of distinct DC subsets.

  2020年10月, Cellular & molecular immunology, 18(6) (6), 1437 - 1449, 英語, 国際誌

  [査読有り]

  研究論文（学術雑誌）


• Nonlinear Optics with Near-Infrared Excitation Enable Real-Time Quantitative Diagnosis of Human Cervical Cancers.

  Takahiro Matsui, Ryo Tamoto, Akio Iwasa, Masafumi Mimura, Seiji Taniguchi, Tetsuo Hasegawa, Takao Sudo, Hiroki Mizuno, Junichi Kikuta, Ichiro Onoyama, Kaoru Okugawa, Mayu Shiomi, Shinya Matsuzaki, Eiichi Morii, Tadashi Kimura, Kiyoko Kato, Yasujiro Kiyota, Masaru Ishii

  Histopathologic analysis through biopsy has been one of the most useful methods for the assessment of malignant neoplasms. However, some aspects of the analysis such as invasiveness, evaluation range, and turnaround time from biopsy to report could be improved. Here, we report a novel method for visualizing human cervical tissue three-dimensionally, without biopsy, fixation, or staining, and with sufficient quality for histologic diagnosis. Near-infrared excitation and nonlinear optics were employed to visualize unstained human epithelial tissues of the cervix uteri by constructing images with third-harmonic generation (THG) and second-harmonic generation (SHG). THG images enabled evaluation of nuclear morphology in a quantitative manner with six parameters after image analysis using deep learning. It was also possible to quantitatively assess intraepithelial fibrotic changes based on SHG images and another deep learning analysis. Using each analytical procedure alone, normal and cancerous tissue were classified quantitatively with an AUC ≥0.92. Moreover, a combinatory analysis of THG and SHG images with a machine learning algorithm allowed accurate classification of three-dimensional image files of normal tissue, intraepithelial neoplasia, and invasive carcinoma with a weighted kappa coefficient of 0.86. Our method enables real-time noninvasive diagnosis of cervical lesions, thus constituting a potential tool to dramatically change early detection. SIGNIFICANCE: This study proposes a novel method for diagnosing cancer using nonlinear optics, which enables visualization of histologic features of living tissues without the need for any biopsy or staining dye.

  2020年09月, Cancer research, 80(17) (17), 3745 - 3754, 英語, 国際誌

  [査読有り]

  研究論文（学術雑誌）


• Development of an intravital imaging system for the synovial tissue reveals the dynamics of CTLA-4 Ig in vivo.

  Tetsuo Hasegawa, Junichi Kikuta, Takao Sudo, Erika Yamashita, Shigeto Seno, Tsutomu Takeuchi, Masaru Ishii

  There have been many attempts to visualize the inflamed joints using multiphoton microscopy. However, due to the hypervascular and multilayered structure of the inflamed synovium, intravital imaging of the deep synovial tissue has been difficult. Here, we established original intravital imaging systems to visualize synovial tissue and pathological osteoclasts at the pannus-bone interface using multiphoton microscopy. Combined with fluorescence-labeling of CTLA-4 Ig, a biological agent used for the treatment of rheumatoid arthritis, we identified that CTLA-4 Ig was distributed predominantly within the inflamed synovium and bound to CX3CR1+ macrophages and CD140a+ fibroblasts 6 h after injection, but not to mature osteoclasts. Intravital imaging of blood and lymphatic vessels in the inflamed synovium further showed that extravasated CTLA-4 Ig was immediately drained through lymphatic vessels under acute arthritic conditions, but the drainage activity was retarded under chronic conditions. These results indicate that this intravital synovial imaging system can serve as a platform for exploring the dynamics of immune cells, osteoclasts, and biological agents within the synovial microenvironment in vivo.

  2020年08月, Scientific reports, 10(1) (1), 13480 - 13480, 英語, 国際誌

  [査読有り]

  研究論文（学術雑誌）


• An Acid-Activatable Fluorescence Probe for Imaging Osteocytic Bone Resorption Activity in Deep Bone Cavities.

  Ryu Hashimoto, Masafumi Minoshima, Junichi Kikuta, Shinya Yari, Steven D Bull, Masaru Ishii, Kazuya Kikuchi

  A rationally designed pH-activatable fluorescent probe (pHocas-RIS) has been used to measure localised pH levels in osteocytic lacunae in bone tissue. Conjugation of the moderate bone-binding drug risedronate to a pH-activatable BODIPY fluorophore enables the probe to penetrate osteocytic lacunae cavities that are embedded deep within the bone matrix. After injection of pHocas-RIS, any osteocytic lacunae caused by bone-resorbing osteocytes cause the probe to fluoresce in vivo, thus allowing imaging by intravital two-photon excitation microscopy. This pH responsive probe enabled the visualization of the bone mineralizing activities of acid producing osteocytes in real time, thus allowing the study of their central role in remodeling the bone-matrix in healthy and disease states.

  Wiley, 2020年08月, Angewandte Chemie (International ed. in English), 英語, 国際誌

  [査読有り]

  研究論文（学術雑誌）


• Thrombomodulin induces anti-inflammatory effects by inhibiting the rolling adhesion of leukocytes in vivo.

  Shino Nishizawa, Junichi Kikuta, Shigeto Seno, Masahiro Kajiki, Ryuichi Tsujita, Hiroki Mizuno, Takao Sudo, Tomoka Ao, Hideo Matsuda, Masaru Ishii

  Thrombomodulin (TM) is an integral membrane protein expressed on the surface of vascular endothelial cells that suppresses blood coagulation. Recent studies have shown that TM exhibits anti-inflammatory effects by inhibiting leukocyte recruitment. However, the actual modes of action of TM in vivo remain unclear. Here, we describe the pharmacological effects of recombinant human soluble TM (TM alfa) on leukocyte dynamics in living mice using intravital imaging techniques. Under control conditions, neutrophils exhibited three distinct types of adhesion behavior in vessels: 1) "non-adhesion", in which cells flowed without vessel adhesion; 2) "rolling adhesion", in which cells transiently interacted with the endothelium; and 3) "tight binding", in which cells bound strongly to the endothelial cells. Compared to control conditions, local lipopolysaccharide stimulation resulted in an increased frequency of rolling adhesion that was not homogeneously distributed on vessel walls but occurred at specific endothelial sites. Under inflammatory conditions, TM alfa, particularly the D1 domain which is a lectin-like region of TM, significantly decreased the frequency of rolling adhesion, but did not influence the number of tight bindings. This was the first study to demonstrate that TM alfa exerts anti-inflammatory effects by inhibiting rolling adhesion of neutrophils to vascular endothelial cells in living mice.

  2020年05月, Journal of pharmacological sciences, 143(1) (1), 17 - 22, 英語, 国内誌

  [査読有り]

  研究論文（学術雑誌）


• In vivo dynamic analysis of BMP-2-induced ectopic bone formation.

  Kunihiko Hashimoto, Takashi Kaito, Masayuki Furuya, Shigeto Seno, Daisuke Okuzaki, Junichi Kikuta, Hiroyuki Tsukazaki, Hideo Matsuda, Hideki Yoshikawa, Masaru Ishii

  2020年03月, Scientific reports, 10(1) (1), 4751 - 4751, 英語, 国際誌

  [査読有り]

  研究論文（学術雑誌）


• 骨芽細胞・破骨細胞は細胞間接触を介して骨リモデリングを調節する

  古家 雅之, 菊田 順一, 瀬尾 茂人, 前田 拓樹, 柏井 将文, 海渡 貴司, 橋本 国彦, 松田 秀雄, 菊地 和也, 石井 優, 吉川 秀樹

  (公社)日本整形外科学会, 2020年03月, 日本整形外科学会雑誌, 94(3) (3), S1099 - S1099, 日本語


• 骨芽細胞・破骨細胞は細胞間接触を介して骨リモデリングを調節する

  古家 雅之, 菊田 順一, 瀬尾 茂人, 前田 拓樹, 柏井 将文, 海渡 貴司, 橋本 国彦, 松田 秀雄, 菊地 和也, 石井 優, 吉川 秀樹

  (一社)日本脊椎脊髄病学会, 2020年03月, Journal of Spine Research, 11(3) (3), 521 - 521, 日本語


• Intravital multiphoton microscopy as a novel tool in the field of immunopharmacology.

  Akito Morimoto, Junichi Kikuta, Masaru Ishii

  Intravital microscopy with multiphoton excitation is a recently developed optical imaging technique for deep tissue imaging without fixation or sectioning, which permits examination of fundamental concepts regarding the dynamic nature of cells under physiological and pathological conditions in living animals. This novel technique also offers exciting opportunities for pharmacological research by providing new platforms for the study of cellular dynamics in response to drugs in vivo. Moreover, fluorescent chemical probes for functional or molecular analysis in single cells in vivo play important roles in pharmacology. For example, we have recently revealed the pharmacodynamic actions of different biological agents for the treatment of rheumatoid arthritis (RA) in vivo by directly visualizing drug-induced cellular behaviors and functions of osteoclasts on bone surfaces. This review focuses on the principles and advantages of intravital imaging for the dissection of pharmacological mechanisms, and discusses how such imaging can contribute to the drug development process, introducing recent trials that evaluated the in vivo pharmacological effects of various agents.

  2020年02月, Pharmacology & therapeutics, 206, 107429 - 107429, 英語, 国際誌

  [査読有り]

  研究論文（学術雑誌）


• Migration arrest of chemoresistant leukemia cells mediated by MRTF-SRF pathway.

  Maho Morimatsu, Erika Yamashita, Shigeto Seno, Takao Sudo, Junichi Kikuta, Hiroki Mizuno, Daisuke Okuzaki, Daisuke Motooka, Masaru Ishii

  Background: Dormant chemotherapy-resistant leukemia cells can survive for an extended period before relapse. Nevertheless, the mechanisms underlying the development of chemoresistance in vivo remain unclear. Methods: Using intravital bone imaging, we characterized the behavior of murine acute myeloid leukemia (AML) cells (C1498) in the bone marrow before and after chemotherapy with cytarabine. Results: Proliferative C1498 cells exhibited high motility in the bone marrow. Cytarabine treatment impaired the motility of residual C1498 cells. However, C1498 cells regained their migration potential after relapse. RNA sequencing revealed that cytarabine treatment promoted MRTF-SRF pathway activation. MRTF inhibition using CCG-203971 augmented the anti-tumor effects of chemotherapy in our AML mouse model, as well as suppressed the migration of chemoresistant C1498 cells. Conclusions: These results provide novel insight into the role of cell migration arrest on the development of chemoresistance in AML, as well as provide a strong rationale for the modulation of cellular motility as a therapeutic target for refractory AML.

  2020年, Inflammation and regeneration, 40, 15 - 15, 英語, 国際誌

  [査読有り]

  研究論文（学術雑誌）


• Mesenchymal stem cells cultured under hypoxic conditions had a greater therapeutic effect on mice with liver cirrhosis compared to those cultured under normal oxygen conditions.

  Yuichi Kojima, Atsunori Tsuchiya, Masahiro Ogawa, Shunsuke Nojiri, Suguru Takeuchi, Takayuki Watanabe, Kenji Nakajima, Yukio Hara, Junji Yamashita, Junichi Kikuta, Masaaki Takamura, Masaru Ishii, Shuji Terai

  Background: Mesenchymal stem cells (MSCs) can be easily expanded. They can be acquired from medical waste such as adipose and umbilical cord tissues, are influenced by culturing conditions, and exert anti-inflammatory, antioxidant, anti-fibrotic, and angiogenic effects. We analyzed the multi-directional effects of MSCs cultured under hypoxic conditions and their underlying mechanisms in the treatment of liver cirrhosis in a mouse model. Methods: Human bone marrow-derived MSCs cultured under hypoxic (5% O2; hypoMSCs) and normoxic (21% O2; norMSCs) conditions were compared by cap analysis of gene expression (CAGE) with or without serum from liver cirrhosis patients. The therapeutic effects of MSCs, including serum liver enzyme induction, fibrosis regression, and hepatic oxidative stress, were evaluated by injecting 1 × 106, 2 × 105, or 4 × 104 MSCs/mouse into the tail veins of mice with carbon tetrachloride (CCl4)-induced liver cirrhosis. Intravital imaging was performed with a two-photon excitation microscope to confirm the various MSC migration paths to the liver. Results: CAGE analysis revealed that the RNA expression levels of prostaglandin E synthase (Ptges) and miR210 were significantly higher in hypoMSCs than in norMSCs. In vivo analysis revealed that both hypoMSCs and norMSCs reduced serum alanine aminotransferase, oxidative stress, and fibrosis compared to that in control mice in a dose-dependent manner. However, hypoMSCs had stronger therapeutic effects than norMSCs. We confirmed this observation by an in vitro study in which hypoMSCs changed macrophage polarity to an anti-inflammatory phenotype via prostaglandin E2 (PGE2) stimulation. In addition, miR210 reduced the rate of hepatocyte apoptosis. Intravital imaging after MSC administration showed that both cell types were primarily trapped in the lungs. Relatively a few hypoMSCs and norMSCs migrated to the liver. There were no significant differences in their distributions. Conclusion: The therapeutic effect of hypoMSCs was mediated by PGE2 and miR210 production and was greater than that of norMSCs. Therefore, MSCs can be manipulated to improve their therapeutic efficacy in the treatment of liver cirrhosis and could potentially serve in effective cell therapy. MSCs produce several factors with multidirectional effects and function as "conducting cells" in liver cirrhosis.

  2019年12月, Regenerative therapy, 11, 269 - 281, 英語, 国際誌

  [査読有り]

  研究論文（学術雑誌）


• Identification of a novel arthritis-associated osteoclast precursor macrophage regulated by FoxM1.

  Tetsuo Hasegawa, Junichi Kikuta, Takao Sudo, Yoshinobu Matsuura, Takahiro Matsui, Szandor Simmons, Kosuke Ebina, Makoto Hirao, Daisuke Okuzaki, Yuichi Yoshida, Atsushi Hirao, Vladimir V Kalinichenko, Kunihiro Yamaoka, Tsutomu Takeuchi, Masaru Ishii

  Osteoclasts have a unique bone-destroying capacity, playing key roles in steady-state bone remodeling and arthritic bone erosion. Whether the osteoclasts in these different tissue settings arise from the same precursor states of monocytoid cells is presently unknown. Here, we show that osteoclasts in pannus originate exclusively from circulating bone marrow-derived cells and not from locally resident macrophages. We identify murine CX3CR1hiLy6CintF4/80+I-A+/I-E+ macrophages (termed here arthritis-associated osteoclastogenic macrophages (AtoMs)) as the osteoclast precursor-containing population in the inflamed synovium, comprising a subset distinct from conventional osteoclast precursors in homeostatic bone remodeling. Tamoxifen-inducible Foxm1 deletion suppressed the capacity of AtoMs to differentiate into osteoclasts in vitro and in vivo. Furthermore, synovial samples from human patients with rheumatoid arthritis contained CX3CR1+HLA-DRhiCD11c+CD80-CD86+ cells that corresponded to mouse AtoMs, and human osteoclastogenesis was inhibited by the FoxM1 inhibitor thiostrepton, constituting a potential target for rheumatoid arthritis treatment.

  2019年12月, Nature immunology, 20(12) (12), 1631 - 1643, 英語, 国際誌

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  研究論文（学術雑誌）


• Single-cell transcriptome analysis of mouse leukocytes in inflammatory stimulation

  Naoki Osato, Hironori Shigeta, Shigeto Seno, Yutaka Uchida, Junichi Kikuta, Masaru Ishii, Hideo Matsuda

  IEEE, 2019年11月, 2019 IEEE International Conference on Bioinformatics and Biomedicine (BIBM), 1229 - 1231, 英語, 国際誌

  [査読有り]

  研究論文（国際会議プロシーディングス）


• Analyzing Leukocyte Migration Trajectories by Deformable Image Matching

  Hironori Shigeta, Shigeto Seno, Shino Nishizawa, Yutaka Uchida, Junichi Kikuta, Masaru Ishii, Hideo Matsuda

  2019年10月, IEEE 19th International Conference on Bioinformatics and Bioengineering (BIBE), 94 - 98, 英語

  [査読有り]

  研究論文（国際会議プロシーディングス）


• In Vivo Multicolor Imaging with Fluorescent Probes Revealed the Dynamics and Function of Osteoclast Proton Pumps.

  Masafumi Minoshima, Junichi Kikuta, Yuta Omori, Shigeto Seno, Riko Suehara, Hiroki Maeda, Hideo Matsuda, Masaru Ishii, Kazuya Kikuchi

  In vivo two-photon fluorescence imaging is a powerful modality to monitor cell dynamics in biomedical studies. To detect protein functions in living animals in real-time, fluorescent probes must show a quick response to the target function in specific tissues. Here, we developed a rhodamine-based small-molecule fluorescent probe called Red-pHocas (red pH-activatable fluorescent probe for osteoclast activity sensing) to reversibly detect the acidic environments for the spatiotemporal analysis of the function of osteoclast proton pumps. The introduction of electron-withdrawing N-alkyl substituents in the rhodamine spirolactam fluorophore remarkably increased the kinetics of the fluorescence response to acidic pHs, which allowed the rapid and reversible monitoring of acidic compartments and the analysis of the dynamics of osteoclast proton pumps during osteoclastic bone resorption. In vivo multicolor two-photon imaging using Red-pHocas in fluorescent reporter mice revealed that bone acidification occurred synchronously with the accumulation of proton pumps onto the bone surfaces. To our knowledge, this is the first study to demonstrate the direct involvement of osteoclast proton pumps in bone acidification under intravital conditions by means of an imaging probe.

  2019年06月, ACS central science, 5(6) (6), 1059 - 1066, 英語, 国際誌

  [査読有り]

  研究論文（学術雑誌）


• Mesenchymal Stem Cells and Induced Bone Marrow-Derived Macrophages Synergistically Improve Liver Fibrosis in Mice.

  Yusuke Watanabe, Atsunori Tsuchiya, Satoshi Seino, Yuzo Kawata, Yuichi Kojima, Shunzo Ikarashi, Philip J Starkey Lewis, Wei-Yu Lu, Junichi Kikuta, Hirokazu Kawai, Satoshi Yamagiwa, Stuart J Forbes, Masaru Ishii, Shuji Terai

  We describe a novel therapeutic approach for cirrhosis using mesenchymal stem cells (MSCs) and colony-stimulating factor-1-induced bone marrow-derived macrophages (id-BMMs) and analyze the mechanisms underlying fibrosis improvement and regeneration. Mouse MSCs and id-BMMs were cultured from mouse bone marrow and their interactions analyzed in vitro. MSCs, id-BMMs, and a combination therapy using MSCs and id-BMMs were administered to mice with CCl4 -induced cirrhosis. Fibrosis regression, liver regeneration, and liver-migrating host cells were evaluated. Administered cell behavior was also tracked by intravital imaging. In coculture, MSCs induced switching of id-BMMs toward the M2 phenotype with high phagocytic activity. In vivo, the combination therapy reduced liver fibrosis (associated with increased matrix metalloproteinases expression), increased hepatocyte proliferation (associated with increased hepatocyte growth factor, vascular endothelial growth factor, and oncostatin M in the liver), and reduced blood levels of liver enzymes, more effectively than MSCs or id-BMMs monotherapy. Intravital imaging showed that after combination cell administration, a large number of id-BMMs, which phagocytosed hepatocyte debris and were retained in the liver for more than 7 days, along with a few MSCs, the majority of which were trapped in the lung, migrated to the fibrotic area in the liver. Host macrophages and neutrophils infiltrated after combination therapy and contributed to liver fibrosis regression and promoted regeneration along with administered cells. Indirect effector MSCs and direct effector id-BMMs synergistically improved cirrhosis along with host cells in mice. These studies pave the way for new treatments for cirrhosis. Stem Cells Translational Medicine 2019;8:271&284.

  2019年03月, Stem cells translational medicine, 8(3) (3), 271 - 284, 英語, 国際誌

  [査読有り]

  研究論文（学術雑誌）


• GPR31-dependent dendrite protrusion of intestinal CX3CR1+ cells by bacterial metabolites.

  Naoki Morita, Eiji Umemoto, Setsuko Fujita, Akio Hayashi, Junichi Kikuta, Ikuo Kimura, Takeshi Haneda, Toshio Imai, Asuka Inoue, Hitomi Mimuro, Yuichi Maeda, Hisako Kayama, Ryu Okumura, Junken Aoki, Nobuhiko Okada, Toshiyuki Kida, Masaru Ishii, Ryusuke Nabeshima, Kiyoshi Takeda

  2019年02月, Nature, 566(7742) (7742), 110 - 114, 英語, 国際誌

  [査読有り]

  研究論文（学術雑誌）


• Imaging the Bone-Immune Cell Interaction in Bone Destruction.

  Tetsuo Hasegawa, Junichi Kikuta, Masaru Ishii

  Bone is a highly dynamic organ that is continuously being remodeled by the reciprocal interactions between bone and immune cells. We have originally established an advanced imaging system for visualizing the in vivo behavior of osteoclasts and their precursors in the bone marrow cavity using two-photon microscopy. Using this system, we found that the blood-enriched lipid mediator, sphingosine-1-phosphate, controlled the migratory behavior of osteoclast precursors. We also developed pH-sensing chemical fluorescent probes to detect localized acidification by bone-resorbing osteoclasts on the bone surface in vivo, and identified two distinct functional states of differentiated osteoclasts, "bone-resorptive" and "non-resorptive." Here, we summarize our studies on the dynamics and functions of bone and immune cells within the bone marrow. We further discuss how our intravital imaging techniques can be applied to evaluate the mechanisms of action of biological agents in inflammatory bone destruction. Our intravital imaging techniques would be beneficial for studying the cellular dynamics in arthritic inflammation and bone destruction in vivo and would also be useful for evaluating novel therapies in animal models of bone-destroying diseases.

  2019年, Frontiers in immunology, 10, 596 - 596, 英語, 国際誌

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  研究論文（学術雑誌）


• Dynamic Analyses of the Short-Term Effects of Different Bisphosphonates Using Intravital Two-Photon Microscopy.

  Junichi Kikuta, Mai Shirazaki, Takao Sudo, Hiroki Mizuno, Akito Morimoto, Riko Suehara, Masafumi Minoshima, Kazuya Kikuchi, Masaru Ishii

  Bisphosphonates are commonly used for the treatment of bone disorders such as osteoporosis; however, the mechanism by which they affect the dynamics of living mature osteoclasts in vivo remains unknown. Here, we describe the short-term effects of different bisphosphonates on controlling the bone resorptive activity of mature osteoclasts in living bone tissues of mice using intravital two-photon microscopy with a pH-sensing chemical fluorescent probe. Three types of nitrogen-containing bisphosphonates, risedronate, alendronate, and minodronate, inhibited osteoclastic acidification during osteoporotic conditions just 12 hours after i.v. injection. Among the three types of drugs, risedronate was the most effective at increasing osteoclast motility and changing the localization of proton pumps, which led to an inhibition of bone resorption. Together, these results demonstrate that the intravital imaging system is a useful tool for evaluating the similarities and differences in currently used antibone resorptive drugs. © 2018 The Authors. JBMR Plus published by Wiley Periodicals, Inc. on behalf of American Society for Bone and Mineral Research.

  2018年11月, JBMR plus, 2(6) (6), 362 - 366, 英語, 国際誌

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  研究論文（学術雑誌）


• Intravital imaging with two-photon microscopy reveals cellular dynamics in the ischeamia-reperfused rat heart.

  Ryohei Matsuura, Shigeru Miyagawa, Satsuki Fukushima, Takasumi Goto, Akima Harada, Yuri Shimozaki, Kazumasa Yamaki, Sho Sanami, Junichi Kikuta, Masaru Ishii, Yoshiki Sawa

  Recent advances in intravital microscopy have provided insight into dynamic biological events at the cellular level in both healthy and pathological tissue. However, real-time in vivo cellular imaging of the beating heart has not been fully established, mainly due to the difficulty of obtaining clear images through cycles of cardiac and respiratory motion. Here we report the successful recording of clear in vivo moving images of the beating rat heart by two-photon microscopy facilitated by cardiothoracic surgery and a novel cardiac stabiliser. Subcellular dynamics of the major cardiac components including the myocardium and its subcellular structures (i.e., nuclei and myofibrils) and mitochondrial distribution in cardiac myocytes were visualised for 4-5 h in green fluorescent protein-expressing transgenic Lewis rats at 15 frames/s. We also observed ischaemia/reperfusion (I/R) injury-induced suppression of the contraction/relaxation cycle and the consequent increase in cell permeability and leukocyte accumulation in cardiac tissue. I/R injury was induced in other transgenic mouse lines to further clarify the biological events in cardiac tissue. This imaging system can serve as an alternative modality for real time monitoring in animal models and cardiological drug screening, and can contribute to the development of more effective treatments for cardiac diseases.

  2018年10月, Scientific reports, 8(1) (1), 15991 - 15991, 英語, 国際誌

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  研究論文（学術雑誌）


• Histone methylation regulator PTIP is required to maintain normal and leukemic bone marrow niches.

  Prosun Das, Kylee J Veazey, Hieu T Van, Saakshi Kaushik, Kevin Lin, Yue Lu, Masaru Ishii, Junichi Kikuta, Kai Ge, Andre Nussenzweig, Margarida A Santos

  The bone is essential for locomotion, calcium storage, and harboring the hematopoietic stem cells (HSCs) that supply the body with mature blood cells throughout life. HSCs reside at the interface of the bone and bone marrow (BM), where active bone remodeling takes place. Although the cellular components of the BM niche have been characterized, little is known about its epigenetic regulation. Here we find that the histone methylation regulator PTIP (Pax interaction with transcription-activation domain protein-1) is required to maintain the integrity of the BM niche by promoting osteoclast differentiation. PTIP directly promotes chromatin changes required for the expression of Pparγ (peroxisome proliferator-activated receptor-γ), a transcription factor essential for osteoclastogenesis. PTIP deletion leads to a drastic reduction of HSCs in the BM and induces extramedullary hematopoiesis. Furthermore, exposure of acute myeloid leukemia cells to a PTIP-deficient BM microenvironment leads to a reduction in leukemia-initiating cells and increased survival upon transplantation. Taken together, our data identify PTIP as an epigenetic regulator of osteoclastogenesis that is required for the integrity of the BM niche to sustain both normal hematopoiesis and leukemia.

  2018年10月, Proceedings of the National Academy of Sciences of the United States of America, 115(43) (43), E10137-E10146 - E10146, 英語, 国際誌

  [査読有り]

  研究論文（学術雑誌）


• The 17,18-epoxyeicosatetraenoic acid-G protein-coupled receptor 40 axis ameliorates contact hypersensitivity by inhibiting neutrophil mobility in mice and cynomolgus macaques.

  Takahiro Nagatake, Yumiko Shiogama, Asuka Inoue, Junichi Kikuta, Tetsuya Honda, Prabha Tiwari, Takayuki Kishi, Atsushi Yanagisawa, Yosuke Isobe, Naomi Matsumoto, Michiko Shimojou, Sakiko Morimoto, Hidehiko Suzuki, So-Ichiro Hirata, Pär Steneberg, Helena Edlund, Junken Aoki, Makoto Arita, Hiroshi Kiyono, Yasuhiro Yasutomi, Masaru Ishii, Kenji Kabashima, Jun Kunisawa

  BACKGROUND: Metabolites of eicosapentaenoic acid exert various physiologic actions. 17,18-Epoxyeicosatetraenoic acid (17,18-EpETE) is a recently identified new class of antiallergic and anti-inflammatory lipid metabolite of eicosapentaenoic acid, but its effects on skin inflammation and the underlying mechanisms remain to be investigated. OBJECTIVE: We evaluated the effectiveness of 17,18-EpETE for control of contact hypersensitivity in mice and cynomolgus macaques. We further sought to reveal underlying mechanisms by identifying the responsible receptor and cellular target of 17,18-EpETE. METHODS: Contact hypersensitivity was induced by topical application of 2,4-dinitrofluorobenzene. Skin inflammation and immune cell populations were analyzed by using flow cytometric, immunohistologic, and quantitative RT-PCR analyses. Neutrophil mobility was examined by means of imaging analysis in vivo and neutrophil culture in vitro. The receptor for 17,18-EpETE was identified by using the TGF-α shedding assay, and the receptor's involvement in the anti-inflammatory effects of 17,18-EpETE was examined by using KO mice and specific inhibitor treatment. RESULTS: We found that preventive or therapeutic treatment with 17,18-EpETE ameliorated contact hypersensitivity by inhibiting neutrophil mobility in mice and cynomolgus macaques. 17,18-EpETE was recognized by G protein-coupled receptor (GPR) 40 (also known as free fatty acid receptor 1) and inhibited chemoattractant-induced Rac activation and pseudopod formation in neutrophils. Indeed, the antiallergic inflammatory effect of 17,18-EpETE was abolished in the absence or inhibition of GPR40. CONCLUSION: 17,18-EpETE inhibits neutrophil mobility through GPR40 activation, which is a potential therapeutic target to control allergic inflammatory diseases.

  2018年08月, The Journal of allergy and clinical immunology, 142(2) (2), 470 - 484, 英語, 国際誌

  [査読有り]

  研究論文（学術雑誌）


• In vivo visualisation of different modes of action of biological DMARDs inhibiting osteoclastic bone resorption.

  Yoshinobu Matsuura, Junichi Kikuta, Yuika Kishi, Tetsuo Hasegawa, Daisuke Okuzaki, Toru Hirano, Masafumi Minoshima, Kazuya Kikuchi, Atsushi Kumanogoh, Masaru Ishii

  OBJECTIVES: Osteoclasts play critical roles in inflammatory bone destruction. Precursor cell migration, cell differentiation, and functional cell activation are all in play. Biological disease-modifying antirheumatic drugs (DMARDs) have been shown to significantly inhibit both bone erosion as well as synovitis, although how such agents reduce osteoclastic bone destructionin vivo has not been fully explained. Here, we used an intravital time-lapse imaging technique to directly visualise mature osteoclasts and their precursors, and explored how different biological DMARDs acted in vivo. METHODS: Lipopolysaccharide (LPS) was injected into the calvarial periosteum of fluorescent reporter mice to induce inflammatory bone destruction. Time-lapse imaging was performed via intravital multiphoton microscopy 5 days after LPS injection. Biological DMARDs, including monoclonal antibodies (mAbs) against the interleukin (IL) 6 receptor (IL-6R) and tumour necrosis factor α (TNFα), or cytotoxic T-lymphocyte-associated protein 4 (CTLA4)-Ig, were intraperitoneally administered at the time of LPS injection. We determined CD80/86 expression levels in mature osteoclasts and their precursors by flow cytometry, quantitative PCR and immunohistochemistry. RESULTS: Of the biologicals tested, anti-IL-6R and anti-TNFα mAbs affected mature osteoclasts and switched bone-resorbing osteoclasts to non-resorbing cells. CTLA4-Ig had no action on mature osteoclasts but mobilised osteoclast precursors, eliminating their firm attachment to bone surfaces. In agreement with these results, CD80/86 (the target molecules of CTLA4-Ig) were prominently expressed only in osteoclast precursor cells, being suppressed during osteoclast maturation. CONCLUSIONS: Intravital imaging revealed that various biological DMARDs acted at specific therapeutic time points during osteoclastic bone destruction, with different efficacies. These results enable us to grasp the real modes of action of drugs, optimising the usage of drug regimens.

  2018年08月, Annals of the rheumatic diseases, 77(8) (8), 1219 - 1225, 英語, 国際誌

  [査読有り]

  研究論文（学術雑誌）


• Chiral cell sliding drives left-right asymmetric organ twisting.

  Mikiko Inaki, Ryo Hatori, Naotaka Nakazawa, Takashi Okumura, Tomoki Ishibashi, Junichi Kikuta, Masaru Ishii, Kenji Matsuno, Hisao Honda

  Polarized epithelial morphogenesis is an essential process in animal development. While this process is mostly attributed to directional cell intercalation, it can also be induced by other mechanisms. Using live-imaging analysis and a three-dimensional vertex model, we identified 'cell sliding,' a novel mechanism driving epithelial morphogenesis, in which cells directionally change their position relative to their subjacent (posterior) neighbors by sliding in one direction. In Drosophila embryonic hindgut, an initial left-right (LR) asymmetry of the cell shape (cell chirality in three dimensions), which occurs intrinsically before tissue deformation, is converted through LR asymmetric cell sliding into a directional axial twisting of the epithelial tube. In a Drosophila inversion mutant showing inverted cell chirality and hindgut rotation, cell sliding occurs in the opposite direction to that in wild-type. Unlike directional cell intercalation, cell sliding does not require junctional remodeling. Cell sliding may also be involved in other cases of LR-polarized epithelial morphogenesis.

  2018年06月, eLife, 7, e32506, 英語, 国際誌

  [査読有り]

  研究論文（学術雑誌）


• Claudin-3 Loss Causes Leakage of Sweat from the Sweat Gland to Contribute to the Pathogenesis of Atopic Dermatitis.

  Kosuke Yamaga, Hiroyuki Murota, Atsushi Tamura, Hirofumi Miyata, Masato Ohmi, Junichi Kikuta, Masaru Ishii, Sachiko Tsukita, Ichiro Katayama

  Elsevier B.V., 2018年06月, The Journal of investigative dermatology, 138(6) (6), 1279 - 1287, 英語, 国際誌

  [査読有り]

  研究論文（学術雑誌）


• エクリン汗腺の構造と動態観察による発汗制御メカニズムの解明

  室田 浩之, 谷 佐起, 山賀 康右, 小野 慧美, 田村 淳, 菊田 順一, 近江 雅人, 月田 早智子, 石井 優, 関口 清俊, 片山 一朗

  (公社)日本皮膚科学会, 2018年05月, 日本皮膚科学会雑誌, 128(5) (5), 1173 - 1173, 日本語

  [査読有り]


• In vivo live imaging of bone cells.

  Hiroki Mizuno, Junichi Kikuta, Masaru Ishii

  There are as many as 200 cell types in the body, and highly sophisticated and varied life phenomena are carried out by cell migration to appropriate places at appropriate times following the appropriate interactions. Recent advances in optical imaging technology using multi-photon excitation microscopy have enabled visualization inside intact bone tissues in living animals without thin sectioning. Using such advanced techniques, the dynamic behaviors of living bone cells on intact bone tissue structures can be elucidated. Here, we focus on recent findings using intravital multi-photon imaging of dynamic biological systems, e.g., bone homeostasis. This novel approach has proven beneficial for understanding the mechanisms underlying the spatiotemporal nature of bone remodeling systems and for evaluating the specific modes of actions of novel drugs currently in development, which will contribute to a new chapter in bone and mineral research.

  2018年04月, Histochemistry and cell biology, 149(4) (4), 417 - 422, 英語, 国際誌

  [査読有り]

  研究論文（学術雑誌）


• In vivo imaging of T cell lymphoma infiltration process at the colon.

  Yoshibumi Ueda, Toshiyuki Ishiwata, Seiichi Shinji, Tomio Arai, Yoko Matsuda, Junko Aida, Naotoshi Sugimoto, Toshiro Okazaki, Junichi Kikuta, Masaru Ishii, Moritoshi Sato

  The infiltration and proliferation of cancer cells in the secondary organs are of great interest, since they contribute to cancer metastasis. However, cancer cell dynamics in the secondary organs have not been elucidated at single-cell resolution. In the present study, we established an in vivo model using two-photon microscopy to observe how infiltrating cancer cells form assemblages from single T-cell lymphomas, EL4 cells, in the secondary organs. Using this model, after inoculation of EL4 cells in mice, we discovered that single EL4 cells infiltrated into the colon. In the early stage, sporadic elongated EL4 cells became lodged in small blood vessels. Real-time imaging revealed that, whereas more than 70% of EL4 cells did not move during a 1-hour observation, other EL4 cells irregularly moved even in small vessels and dynamically changed shape upon interacting with other cells. In the late stages, EL4 cells formed small nodules composed of several EL4 cells in blood vessels as well as crypts, suggesting the existence of diverse mechanisms of nodule formation. The present in vivo imaging system is instrumental to dissect cancer cell dynamics during metastasis in other organs at the single-cell level.

  2018年03月, Scientific reports, 8(1) (1), 3978 - 3978, 英語, 国際誌

  [査読有り]

  研究論文（学術雑誌）


• Direct cell–cell contact between mature osteoblasts and osteoclasts dynamically controls their functions in vivo

  Masayuki Furuya, Junichi Kikuta, Sayumi Fujimori, Shigeto Seno, Hiroki Maeda, Mai Shirazaki, Maki Uenaka, Hiroki Mizuno, Yoriko Iwamoto, Akito Morimoto, Kunihiko Hashimoto, Takeshi Ito, Yukihiro Isogai, Masafumi Kashii, Takashi Kaito, Shinsuke Ohba, Ung-il Chung, Alexander C. Lichtler, Kazuya Kikuchi, Hideo Matsuda, Hideki Yoshikawa, Masaru Ishii

  Bone homeostasis is regulated by communication between bone-forming mature osteoblasts (mOBs) and bone-resorptive mature osteoclasts (mOCs). However, the spatial-temporal relationship and mode of interaction in vivo remain elusive. Here we show, by using an intravital imaging technique, that mOB and mOC functions are regulated via direct cell-cell contact between these cell types. The mOBs and mOCs mainly occupy discrete territories in the steady state, although direct cell-cell contact is detected in spatiotemporally limited areas. In addition, a pH-sensing fluorescence probe reveals that mOCs secrete protons for bone resorption when they are not in contact with mOBs, whereas mOCs contacting mOBs are non-resorptive, suggesting that mOBs can inhibit bone resorption by direct contact. Intermittent administration of parathyroid hormone causes bone anabolic effects, which lead to a mixed distribution of mOBs and mOCs, and increase cell-cell contact. This study reveals spatiotemporal intercellular interactions between mOBs and mOCs affecting bone homeostasis in vivo.

  2018年01月, Nature Communications, 9(1) (1), 300 - 300, 英語, 国際誌

  [査読有り]

  研究論文（学術雑誌）


• [Basis of intravital bone imaging.]

  Kikuta J, Ishii M

  2018年, Clinical calcium, 28(2) (2), 175 - 179

  [査読有り]


• [Intravital bone imaging:osteoclast.]

  Kikuta J, Ishii M

  2018年, Clinical calcium, 28(2) (2), 211 - 216

  [査読有り]


• [The effects of anti-bone-resorptive drugs analyzed by intravital bone imaging.]

  Kikuta J, Ishii M

  2018年, Clinical calcium, 28(2) (2), 237 - 242

  [査読有り]


• [Analysis of the bone anabolic agent by intravital imaging technique.]

  Morimoto A, Kikuta J, Furuya M, Ishii M

  Teriparatide, recombinant human PTH(1-34), is the only anabolic agent widely used for osteoporosis. This drug is thought to promote bone formation by modulating bone remodeling system, although the detailed mechanism remains unclear. Recently, we developed a novel intravital imaging technique to visualize mature osteoclasts and mature osteoblasts simultaneously. By means of this system, we revealed the effect of teriparatide on three-dimensional distribution and cell-to-cell interactions between these cells. Advances in these imaging studies may lead to further understanding of the intercellular network in bone metabolism.

  2018年, Clinical calcium, 28(2) (2), 243 - 248, 日本語, 国内誌

  [査読有り]

  研究論文（学術雑誌）


• [Homeostasis and Disorder of Musculoskeletal System.Cellular dynamics in musculoskeletal system visualized by intravital imaging techniques.]

  Kikuta J, Ishii M

  2018年, Clinical calcium, 28(3) (3), 367 - 371

  [査読有り]


• Bone Imaging: Osteoclast and Osteoblast Dynamics.

  Junichi Kikuta, Masaru Ishii

  Bone is continually remodeled by bone-resorbing osteoclasts and bone-forming osteoblasts. Although it has long been believed that bone homeostasis is tightly regulated by communication between osteoclasts and osteoblasts, the fundamental process and dynamics have remained elusive. To resolve this, we established an intravital bone imaging system using multiphoton microscopy to visualize mature osteoclasts and osteoblasts in living bone.We herein describe the methodology for visualizing the in vivo behavior of bone-resorbing osteoclasts and bone-forming osteoblasts in living bone tissues using intravital multiphoton microscopy. This approach facilitates investigation of cellular dynamics in the pathogenesis of bone-destructive disorders, such as osteoporosis and rheumatoid arthritis in vivo, and would thus be useful for evaluating the efficacy of novel anti-bone-resorptive drugs.

  2018年, Methods in molecular biology (Clifton, N.J.), 1763, 1 - 9, 英語, 国際誌

  [査読有り]

  研究論文（学術雑誌）


• Intravital Imaging of Liver Cell Dynamics.

  Sayaka Matsumoto, Junichi Kikuta, Masaru Ishii

  The liver is a vital organ in the body. It has various essential functions, including detoxification, protein synthesis, and control of infection. Because of its medical importance, liver diseases such as hepatitis and cirrhosis can be crucial for an individual. Exploring dynamics of living cells in the liver would provide the clues for understanding the pathology. However, due to its technical difficulty, few studies have used intravital liver imaging. To resolve this, we have established a novel imaging system for visualizing liver cell dynamics in living animals.Herein we describe the methodology for visualizing the in vivo behavior of liver cells using intravital multiphoton microscopy. This approach will be useful for understanding the pathogenesis of liver disorders, as well as liver biology, in vivo.

  2018年, Methods in molecular biology (Clifton, N.J.), 1763, 137 - 143, 英語, 国際誌

  [査読有り]

  研究論文（学術雑誌）


• Intravital Imaging of the Heart at the Cellular Level Using Two-Photon Microscopy.

  Ryohei Matsuura, Shigeru Miyagawa, Junichi Kikuta, Masaru Ishii, Yoshiki Sawa

  Recent molecular approaches have provided deeper insight on heart failure. However, real-time in vivo cellular dynamics have not been satisfactorily visualized. Here, we present a detailed protocol for in vivo cellular imaging for visualization of the rat heart using two-photon microscopy.

  2018年, Methods in molecular biology (Clifton, N.J.), 1763, 145 - 151, 英語, 国際誌

  [査読有り]

  研究論文（学術雑誌）


• Bone marrow cavity segmentation using graph-cuts with wavelet-based texture feature.

  Hironori Shigeta, Tomohiro Mashita, Junichi Kikuta, Shigeto Seno, Haruo Takemura, Masaru Ishii, Hideo Matsuda

  2017年10月, Journal of bioinformatics and computational biology, 15(5) (5), 1740004 - 1740004, 英語, 国際誌

  [査読有り]

  研究論文（学術雑誌）


• Non-labeling multiphoton excitation microscopy as a novel diagnostic tool for discriminating normal tissue and colorectal cancer lesions.

  Takahiro Matsui, Hiroki Mizuno, Takao Sudo, Junichi Kikuta, Naotsugu Haraguchi, Jun-Ichiro Ikeda, Tsunekazu Mizushima, Hirofumi Yamamoto, Eiichi Morii, Masaki Mori, Masaru Ishii

  Multiphoton excitation microscopy (MPM) is regarded as an effective tool that enables the visualization of deep regions within living tissues and organs, with little damage. Here, we report novel non-labeling MPM (NL-MPM) imaging of fresh human colorectal mucosa, which is useful for discriminating cancer lesions from normal tissues quantitatively without any need for resection, fixation, or staining. Using NL-MPM, we visualized three components in human colorectal mucosa, epithelial cells, immune cells, and basement membranes, based on their characteristic patterns of fluorescence. These patterns are characterized by the different auto-fluorescence properties of nicotinamide adenine dinucleotide, nicotinamide adenine dinucleotide phosphate, and flavin adenine dinucleotide and from second harmonic generation (SHG). NL-MPM images were at least as informative to pathologists as were 'conventional' images of fixed tissue sections stained with hematoxylin and eosin. Additionally, two quantitative parameters extracted from NL-MPM images - the nucleus diameter (index N) and the intensity of SHG in the basement membrane (index S) - rendered it possible to diagnose cancer regions effectively. In conclusion, NL-MPM is a novel, promising method for real-time clinical diagnosis of colorectal cancers, and is associated with minimal invasiveness.

  2017年07月, Scientific reports, 7(1) (1), 6959 - 6959, 英語, 国際誌

  [査読有り]

  研究論文（学術雑誌）


• Cell competition with normal epithelial cells promotes apical extrusion of transformed cells through metabolic changes.

  Shunsuke Kon, Kojiro Ishibashi, Hiroto Katoh, Sho Kitamoto, Takanobu Shirai, Shinya Tanaka, Mihoko Kajita, Susumu Ishikawa, Hajime Yamauchi, Yuta Yako, Tomoko Kamasaki, Tomohiro Matsumoto, Hirotaka Watanabe, Riku Egami, Ayana Sasaki, Atsuko Nishikawa, Ikumi Kameda, Takeshi Maruyama, Rika Narumi, Tomoko Morita, Yoshiteru Sasaki, Ryosuke Enoki, Sato Honma, Hiromi Imamura, Masanobu Oshima, Tomoyoshi Soga, Jun-Ichi Miyazaki, Michael R Duchen, Jin-Min Nam, Yasuhito Onodera, Shingo Yoshioka, Junichi Kikuta, Masaru Ishii, Masamichi Imajo, Eisuke Nishida, Yoichiro Fujioka, Yusuke Ohba, Toshiro Sato, Yasuyuki Fujita

  2017年05月, Nature cell biology, 19(5) (5), 530 - 541, 英語, 国際誌

  [査読有り]

  研究論文（学術雑誌）


• A bone marrow cavity segmentation method using wavelet-based texture feature

  Hironori Shigeta, Tomohiro Mashita, Junichi Kikuta, Shigeto Seno, Haruo Takemura, Hideo Matsuda, Masaru Ishii

  Institute of Electrical and Electronics Engineers Inc., 2017年04月, Proceedings - International Conference on Pattern Recognition, 2133 - 2138, 英語

  [査読有り]

  研究論文（国際会議プロシーディングス）


• Roles for Cell-Cell Adhesion and Contact in Obesity-Induced Hepatic Myeloid Cell Accumulation and Glucose Intolerance.

  Yasutaka Miyachi, Kyoichiro Tsuchiya, Chikara Komiya, Kumiko Shiba, Noriko Shimazu, Shinobu Yamaguchi, Michiyo Deushi, Mizuko Osaka, Kouji Inoue, Yuta Sato, Sayaka Matsumoto, Junichi Kikuta, Kenjiro Wake, Masayuki Yoshida, Masaru Ishii, Yoshihiro Ogawa

  2017年03月, Cell reports, 18(11) (11), 2766 - 2779, 英語, 国際誌

  [査読有り]

  研究論文（学術雑誌）


• [Update on recent progress in vitamin D research. The effects of vitamin D in autoinflammatory diseases.]

  Ao T, Kikuta J, Ishii M

  2017年, Clinical calcium, 27(11) (11), 1551 - 1559

  [査読有り]


• [Imaging of bone and joint destruction].

  Junichi Kikuta, Masaru Ishii

  Osteoclasts are bone-resorbing giant polykaryons that differentiate from mononuclear macrophage/monocyte-lineage hematopoietic precursors. We have originally established an advanced imaging system for visualizing in vivo behavior of osteoclasts and their precursors with intravital two-photon microscopy. By means of the system, we found that sphingosine-1-phosphate, a lipid mediator enriched in blood, controlled the migratory behavior of osteoclast precursors. We also developed pH-sensing chemical fluorescent probes to detect localized acidification by bone-resorbing osteoclasts on the bone surface in vivo, and identified two distinct functional states of differentiated osteoclasts, 'bone-resorptive' and 'non-resorptive'. In this review, we summarize our recent studies on the dynamics and functions of osteoclasts. Our intravital imaging techniques would be beneficial for studying the cellular dynamics in arthritic inflammation and bone destruction in vivo and would thus be useful for evaluating novel therapies targeting aspects of osteoclast dynamics in patients with bone-destructive diseases.

  2017年, Nihon Rinsho Men'eki Gakkai kaishi = Japanese journal of clinical immunology, 40(5) (5), 344 - 351, 日本語, 国内誌

  [査読有り]

  研究論文（学術雑誌）


• 模様特徴量とグラフカットを利用した生体骨組織の骨髄腔の領域分割

  繁田 浩功, 間下 以大, 菊田 順一, 瀬尾 茂人, 石井 優, 松田 秀雄

  2016年09月, 日本語

  研究論文（その他学術会議資料等）


• Prickle1 promotes focal adhesion disassembly in cooperation with the CLASP-LL5β complex in migrating cells.

  Lim BC, Matsumoto S, Yamamoto H, Mizuno H, Kikuta J, Ishii M, Kikuchi A

  Prickle is known to be involved in planar cell polarity, including convergent extension and cell migration; however, the detailed mechanism by which Prickle regulates cellular functions is not well understood. Here, we show that Prickle1 regulates front-rear polarization and migration of gastric cancer MKN1 cells. Prickle1 preferentially accumulated at the cell retraction site in close proximity to paxillin at focal adhesions. Prickle1 dynamics correlated with those of paxillin during focal adhesion disassembly. Furthermore, Prickle1 was required for focal adhesion disassembly. CLASPs (of which there are two isoforms, CLASP1 and CLASP2, in mammals) and LL5β (also known as PHLDB2) have been reported to form a complex at cell edges and to control microtubule-dependent focal adhesion disassembly. Prickle1 was associated with CLASPs and LL5β, and was required for the LL5β-dependent accumulation of CLASPs at the cell edge. Knockdown of CLASPs and LL5β suppressed Prickle1-dependent cell polarization and migration. Prickle1 localized to the membrane through its farnesyl moiety, and the membrane localization was necessary for Prickle1 to regulate migration, to bind to CLASPs and LL5β, and to promote microtubule targeting of focal adhesions. Taken together, these results suggest that Prickle1 promotes focal adhesion disassembly during the retraction processes of cell polarization and migration.

  The Company of Biologists, 2016年08月, Journal of cell science, 129(16) (16), 3115 - 29, 英語, 国際誌

  [査読有り]

  研究論文（学術雑誌）


• Cell-cycle-controlled radiation therapy was effective for treating a murine malignant melanoma cell line in vitro and in vivo.

  Keisuke Otani, Yoko Naito, Yukako Sakaguchi, Yuji Seo, Yutaka Takahashi, Junichi Kikuta, Kazuhiko Ogawa, Masaru Ishii

  2016年08月, Scientific reports, 6, 30689 - 30689, 英語, 国際誌

  [査読有り]

  研究論文（学術雑誌）


• Real-time intravital imaging of pH variation associated with osteoclast activity.

  Hiroki Maeda, Toshiyuki Kowada, Junichi Kikuta, Masayuki Furuya, Mai Shirazaki, Shin Mizukami, Masaru Ishii, Kazuya Kikuchi

  2016年08月, Nature chemical biology, 12(8) (8), 579 - 85, 英語, 国際誌

  [査読有り]

  研究論文（学術雑誌）


• 脂肪組織炎症におけるS100A8の意義

  前田 法一, 堰本 亮平, 福田 士郎, 對馬 佑, 松田 圭介, 森 卓也, 中辻 秀朗, 西澤 均, 岸田 堅, 菊田 順一, 舞島 弓子, 船橋 徹, 石井 優, 下村 伊一郎

  (一社)日本内分泌学会, 2016年04月, 日本内分泌学会雑誌, 92(1) (1), 214 - 214, 日本語


• Intravital imaging of the cellular dynamics of LysM-positive cells in a murine corneal suture model.

  Mayumi Ueta, Ayaka Koga, Junichi Kikuta, Keiko Yamada, Sachi Kojima, Katsuhiko Shinomiya, Masaru Ishii, Shigeru Kinoshita

  2016年03月, The British journal of ophthalmology, 100(3) (3), 432 - 5, 英語, 国際誌

  [査読有り]

  研究論文（学術雑誌）


• Visualization of Intravital Immune Cell Dynamics After Conjunctival Surgery Using Multiphoton Microscopy.

  Sachi Kojima, Toshihiro Inoue, Junichi Kikuta, Masayuki Furuya, Ayaka Koga, Tomokazu Fujimoto, Mayumi Ueta, Shigeru Kinoshita, Masaru Ishii, Hidenobu Tanihara

  PURPOSE: To visualize intravital immune cell dynamics in the subconjunctival tissue during the wound-healing process using multiphoton microscopy. METHODS: Gene-targeted mice expressing enhanced green fluorescent protein under the control of the endogenous lysozyme M promoter (LysM-eGFP mice) were anesthetized with isoflurane, and injured by a 10-0 nylon conjunctival suture. Vessels were visualized by intravenous injection of 70 kDa rhodamine-conjugated dextran. Using a multiphoton microscope, the three-dimensional images of the subconjunctival tissue were acquired every minute for 20 minutes before and 0.5, 3, 6, and 72 hours after injury. Raw imaging data were processed for four-dimensional images and analyzed for the number and the velocity of the LysM-eGFP-positive cells using Imaris software. RESULTS: The intravital LysM-eGFP-positive cells and the red-labeled vessels were successfully visualized using a multiphoton microscope. The conjunctival and scleral collagen fibers were detected as secondary harmonic generation signals, which were colored blue. Compared with mice without injury, the number of LysM-eGFP-positive cells in the subconjunctival tissue after conjunctival surgery increased in a time-dependent manner. The cell velocities significantly increased until 3 hours after surgery (5.9 ± 3.2 μm/min; P < 0.0001) and the elevated level was sustained until 72 hours after injury (5.9 ± 3.3 μm/min). CONCLUSION: This is the first report to visualize and evaluate intravital cellular dynamics during inflammation in the subconjunctival tissue using multiphoton microscopy. This technique may be a useful tool to characterize the molecular mechanisms of the wound-healing process after various ocular injuries, such as glaucoma surgery.

  2016年03月, Investigative ophthalmology & visual science, 57(3) (3), 1207 - 12, 英語, 国際誌

  [査読有り]

  研究論文（学術雑誌）


• Fibroblastic reticular cell-derived lysophosphatidic acid regulates confined intranodal T-cell motility.

  Akira Takeda, Daichi Kobayashi, Keita Aoi, Naoko Sasaki, Yuki Sugiura, Hidemitsu Igarashi, Kazuo Tohya, Asuka Inoue, Erina Hata, Noriyuki Akahoshi, Haruko Hayasaka, Junichi Kikuta, Elke Scandella, Burkhard Ludewig, Satoshi Ishii, Junken Aoki, Makoto Suematsu, Masaru Ishii, Kiyoshi Takeda, Sirpa Jalkanen, Masayuki Miyasaka, Eiji Umemoto

  2016年02月, eLife, 5, e10561, 英語, 国際誌

  [査読有り]

  研究論文（学術雑誌）


• Intravital imaging of Ca(2+) signals in lymphocytes of Ca(2+) biosensor transgenic mice: indication of autoimmune diseases before the pathological onset.

  Soichiro Yoshikawa, Takako Usami, Junichi Kikuta, Masaru Ishii, Tetsuo Sasano, Koji Sugiyama, Tetsushi Furukawa, Eiji Nakasho, Hiroshi Takayanagi, Thomas F Tedder, Hajime Karasuyama, Atsushi Miyawaki, Takahiro Adachi

  2016年01月, Scientific reports, 6, 18738 - 18738, 英語, 国際誌

  [査読有り]

  研究論文（学術雑誌）


• Dynamic analysis of immune inflammation and bone destruction by intravital imaging.

  Junichi Kikuta, Masaru Ishii

  Rapid development of fluorescent imaging techniques enables us to understand cellular dynamics in vivo. We have originally established an advanced imaging system for visualizing living bone tissues with intravital two-photon microscopy. By means of the system, we have recently succeeded in visualization of the in vivo behavior of living mature osteoclasts on the bone surface, and identified different functional subsets of osteoclasts in terms of their motility and function, i.e., 'static - bone resorptive' and 'moving - non resorptive'. Pathological conditions changed the composition of these populations as well as the total number of mature osteoclasts. We also found that RANKL-bearing Th17 cells could control bone resorption of mature osteoclasts, demonstrating novel actions of Th17. Furthermore, we have also developed the imaging system to visualize bone destruction by osteoclasts in arthritic joints using intravital two-photon microscopy. In this review, we summarize the latest data of intravital imaging of osteoclast dynamics, and also discuss its further application.

  2016年, Nihon Rinsho Men'eki Gakkai kaishi = Japanese journal of clinical immunology, 39(2) (2), 124 - 9, 日本語, 国内誌

  [査読有り]

  研究論文（学術雑誌）


• Adipose tissue macrophages induce PPARγ-high FOXP3(+) regulatory T cells.

  Onodera T, Fukuhara A, Jang MH, Shin J, Aoi K, Kikuta J, Otsuki M, Ishii M, Shimomura I

  Numerous regulatory T cells (Tregs) are present in adipose tissues compared with other lymphoid or non-lymphoid tissues. Adipose Tregs regulate inflammatory state and insulin sensitivity. However, the mechanism that maintains Tregs in adipose tissue remains unclear. Here, we revealed the contribution of adipose tissue macrophages (ATMs) to the induction and proliferation of adipose Tregs. ATMs isolated from mice under steady state conditions induced Tregs with high expression of PPARγ compared with splenic dendritic cells in vitro. Furthermore, ATMs from obese mice prompted the differentiation of PPARγ low Tregs. Adoptive transfer of ATMs induced differentiation and proliferation of Tregs, whereas depletion of ATMs by clodronate-liposome resulted in reduction of adipose Tregs, in vivo. Deficiency of anti-inflammatory adipocytokine, Adipoq, resulted in small proportions of ATMs and adipose Tregs without alteration of other immune cells in vivo. Therefore, these data suggest that the abundance of Tregs in adipose tissue could be partly attributed to the ability of ATMs to induce PPARγ-expressing Tregs.

  2015年11月, Scientific reports, 5, 16801 - 16801, 英語, 国際誌

  [査読有り]

  研究論文（学術雑誌）


• Oxysterols and EBI2 promote osteoclast precursor migration to bone surfaces and regulate bone mass homeostasis.

  Erin Nevius, Flavia Pinho, Meera Dhodapkar, Huiyan Jin, Kristina Nadrah, Mark C Horowitz, Junichi Kikuta, Masaru Ishii, João P Pereira

  Bone surfaces attract hematopoietic and nonhematopoietic cells, such as osteoclasts (OCs) and osteoblasts (OBs), and are targeted by bone metastatic cancers. However, the mechanisms guiding cells toward bone surfaces are essentially unknown. Here, we show that the Gαi protein-coupled receptor (GPCR) EBI2 is expressed in mouse monocyte/OC precursors (OCPs) and its oxysterol ligand 7α,25-dihydroxycholesterol (7α,25-OHC) is secreted abundantly by OBs. Using in vitro time-lapse microscopy and intravital two-photon microscopy, we show that EBI2 enhances the development of large OCs by promoting OCP motility, thus facilitating cell-cell interactions and fusion in vitro and in vivo. EBI2 is also necessary and sufficient for guiding OCPs toward bone surfaces. Interestingly, OCPs also secrete 7α,25-OHC, which promotes autocrine EBI2 signaling and reduces OCP migration toward bone surfaces in vivo. Defective EBI2 signaling led to increased bone mass in male mice and protected female mice from age- and estrogen deficiency-induced osteoporosis. This study identifies a novel pathway involved in OCP homing to the bone surface that may have significant therapeutic potential.

  2015年10月, The Journal of experimental medicine, 212(11) (11), 1931 - 46, 英語, 国際誌

  [査読有り]

  研究論文（学術雑誌）


• Trafficking of Osteoclast Precursors

  Junichi Kikuta, Erin Nevius, Masaru Ishii, João P. Pereira

  Elsevier Inc., 2015年10月, Osteoimmunology: Interactions of the Immune and Skeletal Systems: Second Edition, 25 - 40, 英語

  [査読有り]

  論文集(書籍)内論文


• 脂肪組織慢性炎症のinitial triggerとしてのS100A8の意義

  前田 法一, 堰本 亮平, 福田 士郎, 對馬 佑, 松田 圭介, 森 卓也, 中辻 秀朗, 西澤 均, 岸田 堅, 菊田 順一, 舞島 弓子, 船橋 徹, 石井 優, 下村 伊一郎

  (一社)日本肥満学会, 2015年09月, 肥満研究, 21(Suppl.) (Suppl.), 175 - 175, 日本語


• コンポーネントツリーを用いたグローバルデータアソシエーションによる細胞追跡手法

  藏重昂平, 瀬尾茂人, 間下以大, 菊田順一, 竹中要一, 石井優, 松田秀雄

  2015年06月, 日本語

  研究論文（その他学術会議資料等）


• [Frontiers in Live Bone Imaging Researches. Intravital imaging of osteoclast dynamics].

  Junichi Kikuta, Masaru Ishii

  Osteoclasts are bone-resorbing giant polykaryons that differentiate from mononuclear macrophage/monocyte-lineage hematopoietic precursors. Upon the stimulation of essential factors such as M-CSF and RANKL, osteoclast precursor monocytes attach to the bone surface ( "migration" ), fuse with each other to form giant cells ( "differentiation" ) and mediate bone resorption ( "function" ). To reveal the regulatory mechanism of these three dynamic steps of osteoclastic activity, we have originally established an advanced imaging system for visualizing living bone tissues with intravital multiphoton microscopy. By means of the system, we have recently succeeded in visualization of osteoclast migration, differentiation, and function in living bone tissues in vivo. In this review we summarize the latest data of intravital imaging of osteoclast dynamics, and discuss novel lines of osteoclast-targeted therapies that will impact future treatment of bone destructive diseases.

  2015年06月, Clinical calcium, 25(6) (6), 815 - 22, 日本語, 国内誌

  [査読有り]

  研究論文（学術雑誌）


• [Intravital multiphoton imaging of live bone tissues revealing hematopoietic cell dynamics in vivo].

  Hiroki Mizuno, Junichi Kikuta, Masayuki Furuya, Masaru Ishii

  During the last decade, multiphoton fluorescent microscopy has launched a new era in the field of biology. By using this advanced imaging technique we have established a system for visualizing the in situ behaviors of a diversity of living cells within intact tissues and organs. Among them we succeeded in visualizing the various dynamic phenomena within bones, a mysterious organ wherein various hematopoietic and immune cells are produced and functioning. However, these cell types are poorly understood with conventional methodologies such as histological analyses of decalcified bone sections. We have thus far focused on the behavior of osteoclasts, a type of specialized macrophage contributing to physiological turnover of bone tissues as well as pathological bone destruction, and have revealed novel mechanisms controlling the migration and function of osteoclasts in situ. Functional coupling between bone-destroying osteoclasts and bone-replenishing osteoblasts could also be visualized demonstrating a genuine mode of 'cross-talk' among these cell types. Furthermore, we have recently imaged in situ behaviors of different hematopoietic cells in bone marrow. Herein we present our latest data on cellular dynamics in the bone cavity, and discuss the applications of this novel methodology for future studies in the field of hematology.

  2015年06月, [Rinsho ketsueki] The Japanese journal of clinical hematology, 56(6) (6), 594 - 600, 日本語, 国内誌

  [査読有り]

  研究論文（学術雑誌）


• Intravital bone imaging by two-photon excitation microscopy to identify osteocytic osteolysis in vivo.

  Hiroshige Sano, Junichi Kikuta, Masayuki Furuya, Naoki Kondo, Naoto Endo, Masaru Ishii

  2015年05月, Bone, 74, 134 - 9, 英語, 国際誌

  [査読有り]

  研究論文（学術雑誌）


• Visualized macrophage dynamics and significance of S100A8 in obese fat.

  Ryohei Sekimoto, Shiro Fukuda, Norikazu Maeda, Yu Tsushima, Keisuke Matsuda, Takuya Mori, Hideaki Nakatsuji, Hitoshi Nishizawa, Ken Kishida, Junichi Kikuta, Yumiko Maijima, Tohru Funahashi, Masaru Ishii, Iichiro Shimomura

  Chronic low-grade inflammation of adipose tissue plays a crucial role in the pathophysiology of obesity. Immunohistological microscopic analysis in obese fat tissue has demonstrated the infiltration of several immune cells such as macrophages, but dynamics of immune cells have not been fully elucidated and clarified. Here, by using intravital multiphoton imaging technique, to our knowledge for the first time, we analyzed and visualized the inflammatory processes in adipose tissue under high-fat and high-sucrose (HF/HS) diet with lysozyme M-EGFP transgenic (LysM(EGFP)) mice whose EGFP was specifically expressed in the myelomonocytic lineage. Mobility of LysM(EGFP)-positive macrophages was shown to be activated just 5 d after HF/HS diet, when the distinct hypertrophy of adipocytes and the accumulation of macrophages still have not become prominent. Significant increase of S100A8 was detected in mature adipocyte fraction just 5 d after HF/HS diet. Recombinant S100A8 protein stimulated chemotactic migration in vitro and in vivo, as well as induced proinflammatory molecules, both macrophages and adipocytes, such as TNF-α and chemokine (C-C motif) ligand 2. Finally, an antibody against S100A8 efficiently suppressed the HF/HS diet-induced initial inflammatory change, i.e., increased mobilization of adipose LysM(EGFP)-positive macrophages, and ameliorated HF/HS diet-induced insulin resistance. In conclusion, time-lapse intravital multiphoton imaging of adipose tissues identified the very early event exhibiting increased mobility of macrophages, which may be triggered by increased expression of adipose S100A8 and results in progression of chronic inflammation in situ.

  2015年04月, Proceedings of the National Academy of Sciences of the United States of America, 112(16) (16), E2058-66 - 66, 英語, 国際誌

  [査読有り]

  研究論文（学術雑誌）


• RFPL4A increases the G1 population and decreases sensitivity to chemotherapy in human colorectal cancer cells.

  Atsushi Naito, Hirofumi Yamamoto, Yoshinori Kagawa, Yoko Naito, Daisuke Okuzaki, Keisuke Otani, Yoriko Iwamoto, Sakae Maeda, Junichi Kikuta, Keizo Nishikawa, Mamoru Uemura, Junichi Nishimura, Taishi Hata, Ichiro Takemasa, Tsunekazu Mizushima, Hideshi Ishii, Yuichiro Doki, Masaki Mori, Masaru Ishii

  Cell cycle-arrested cancer cells are resistant to conventional chemotherapy that acts on the mitotic phases of the cell cycle, although the molecular mechanisms involved in halting cell cycle progression remain unclear. Here, we demonstrated that RFPL4A, an uncharacterized ubiquitin ligase, induced G1 retention and thus conferred decreased sensitivity to chemotherapy in the human colorectal cancer cell line, HCT116. Long term time lapse observations in HCT116 cells bearing a "fluorescence ubiquitin-based cell cycle indicator" identified a characteristic population that is viable but remains in the G1 phase for an extended period of time (up to 56 h). Microarray analyses showed that expression of RFPL4A was significantly up-regulated in these G1-arrested cells, not only in HCT116 cells but also in other cancer cell lines, and overexpression of RFPL4A increased the G1 population and decreased sensitivity to chemotherapy. However, knockdown of RFPL4A expression caused the cells to resume mitosis and induced their susceptibility to anti-cancer drugs in vitro and in vivo. These results indicate that RFPL4A is a novel factor that increases the G1 population and decreases sensitivity to chemotherapy and thus may be a promising therapeutic target for refractory tumor conditions.

  2015年03月, The Journal of biological chemistry, 290(10) (10), 6326 - 37, 英語, 国際誌

  [査読有り]

  研究論文（学術雑誌）


• A Bone Marrow Recognition Method for Bone Tissue Images using Wavelet Transform

  Hironori Shigeta, Tomohiro Mashita, Takeshi Kaneko, Junichi Kikuta, Shigeto Senoo, Haruo Takemura, Hideo Matsuda, Masaru Ishii

  2015年03月, 英語

  研究論文（大学，研究機関等紀要）


• [Current Topics on Vitamin D. The effects of vitamin D on the immune system].

  Junichi Kikuta, Masaru Ishii

  Various kinds of immune cells-including macrophages, dendritic cells, T cells and B cells- express the vitamin D receptor and 1α-hydroxylase (CYP27B1), the enzyme necessary for the conversion of circulating 25-hydroxyvitamin D into its active form, 1,25-dihydroxyvitamin D. It suggests that vitamin D has a regulatory role on innate and adaptive immune responses. Vitamin D has been recently shown to promote antimicrobial responses through the production of antibacterial peptides, and stimulation of the autophagic activity in macrophages. Recent epidemiological evidence indicates a significant association between vitamin D deficiency and an increased incidence of several infectious diseases. Here, we review the essential roles of vitamin D in modulating the immune system and discuss the protective effects of vitamin D supplementation in diverse infections.

  2015年03月, Clinical calcium, 25(3) (3), 359 - 65, 日本語, 国内誌

  [査読有り]

  研究論文（学術雑誌）


• グラフカットを用いた骨髄腔画像の領域分割

  宇佐見 潤, 繁田 浩功, 間下 以大, 黒田 嘉宏, 菊田 順一, 瀬尾 茂人, 石井 優, 松田 秀雄, 竹村 治雄

  2015年03月, 情報処理学会論文誌 数理モデル化と応用, 8(1) (1), 18 - 27, 日本語

  [査読有り]

  研究論文（学術雑誌）


• Transplanted bone marrow-derived circulating PDGFRα+ cells restore type VII collagen in recessive dystrophic epidermolysis bullosa mouse skin graft.

  Iinuma S, Aikawa E, Tamai K, Fujita R, Kikuchi Y, Chino T, Kikuta J, McGrath JA, Uitto J, Ishii M, Iizuka H, Kaneda Y

  Recessive dystrophic epidermolysis bullosa (RDEB) is an intractable genetic blistering skin disease in which the epithelial structure easily separates from the underlying dermis because of genetic loss of functional type VII collagen (Col7) in the cutaneous basement membrane zone. Recent studies have demonstrated that allogeneic bone marrow transplantation (BMT) ameliorates the skin blistering phenotype of RDEB patients by restoring Col7. However, the exact therapeutic mechanism of BMT in RDEB remains unclear. In this study, we investigated the roles of transplanted bone marrow-derived circulating mesenchymal cells in RDEB (Col7-null) mice. In wild-type mice with prior GFP-BMT after lethal irradiation, lineage-negative/GFP-positive (Lin(-)/GFP(+)) cells, including platelet-derived growth factor receptor α-positive (PDGFRα(+)) mesenchymal cells, specifically migrated to skin grafts from RDEB mice and expressed Col7. Vascular endothelial cells and follicular keratinocytes in the deep dermis of the skin grafts expressed SDF-1α, and the bone marrow-derived PDGFRα(+) cells expressed CXCR4 on their surface. Systemic administration of the CXCR4 antagonist AMD3100 markedly decreased the migration of bone marrow-derived PDGFRα(+) cells into the skin graft, resulting in persistent epidermal detachment with massive necrosis and inflammation in the skin graft of RDEB mice; without AMD3100 administration, Col7 was significantly supplemented to ameliorate the pathogenic blistering phenotype. Collectively, these data suggest that the SDF1α/CXCR4 signaling axis induces transplanted bone marrow-derived circulating PDGFRα(+) mesenchymal cells to migrate and supply functional Col7 to regenerate RDEB skin.

  2015年02月, Journal of immunology (Baltimore, Md. : 1950), 194(4) (4), 1996 - 2003, 英語, 国際誌

  [査読有り]

  研究論文（学術雑誌）


• 生体骨組織における骨髄腔画像のウェーブレット変換を用いた骨髄腔領域の認識手法

  繁田 浩功, 間下 以大, 金子 雄, 菊田 順一, 瀬尾 茂人, 竹村 治雄, 松田 秀雄, 石井 優

  生体イメージング技術の向上により生体内の細胞の動態を動画像として観察が可能となり,疾病のメカニズム解明や創薬等への応用が期待されている.これらの応用のためには,細胞画像から特定領域を抽出したり,細胞の特定の動きを検出する必要がある.また,膨大な数の画像に対して一定の基準で領域分割や細胞の検出を行うためには計算機での処理が必要である.本稿では,その一例となる二光子励起顕微鏡を用いた骨髄腔画像の骨髄腔の領域分割手法を検討する.このような画像を解析する際には,時系列画像の時間情報を必要としない認識手法を確立することが好ましい.そのため,本研究では骨髄腔画像に映り込む染まり方への模様に着目し,ウェーブレット変換を用いて特徴量の検出を行い,骨髄腔の認識を行う方法を検討する.本稿で提案した手法を用いることで,対象画像においておおむね抽出に成功した例を確認した.

  一般社団法人情報処理学会, 2015年01月, 情報処理学会研究報告. CVIM, [コンピュータビジョンとイメージメディア], 2015(42) (42), 1 - 5, 日本語

  研究論文（その他学術会議資料等）


• Sweat, the driving force behind normal skin: an emerging perspective on functional biology and regulatory mechanisms.

  Hiroyuki Murota, Saki Matsui, Emi Ono, Akiko Kijima, Junichi Kikuta, Masaru Ishii, Ichiro Katayama

  The various symptoms associated with excessive or insufficient perspiration can significantly reduce a patient's quality of life. If a versatile and minimally invasive method could be established for returning sweat activity to normalcy, there is no question that it could be used in the treatment of many diseases that are believed to involve perspiration. For this reason, based on an understanding of the sweat-gland control function and sweat activity, it was necessary to conduct a comprehensive search for the factors that control sweating, such as the central and peripheral nerves that control sweat-gland function, the microenvironment surrounding the sweat glands, and lifestyle. We focused on the mechanism by which atopic dermatitis leads to hypohidrosis and confirmed that histamine inhibits acetylcholinergic sweating. Acetylcholine promotes the phosphorylation of glycogen synthesis kinase 3β (GSK3β) in the sweat-gland secretory cells and leads to sensible perspiration. By suppressing the phosphorylation of GSK3β, histamine inhibits the movement of sweat from the sweat-gland secretory cells through the sweat ducts, which could presumably be demonstrated by dynamic observations of the sweat glands using two-photon microscopy. It is expected that the discovery of new factors that control sweat-gland function can contribute to the treatment of diseases associated with dyshidrosis.

  2015年01月, Journal of dermatological science, 77(1) (1), 3 - 10, 英語, 国際誌

  [査読有り]

  研究論文（学術雑誌）


• 脂肪組織慢性炎症における免疫細胞の動態変化の解析

  堰本 亮平, 前田 法一, 對馬 佑, 松田 圭介, 森 卓也, 福田 士郎, 中辻 秀朗, 西澤 均, 岸田 堅, 菊田 順一, 船橋 徹, 石井 優, 下村 伊一郎

  (一社)日本肥満学会, 2014年10月, 肥満研究, 20(Suppl.) (Suppl.), 168 - 168, 日本語


• Improvement approach of cell tracking accuracy by using inter-frame information

  Kohei Kurashige, Shigeto Seno, Tomohiro Mashita, Junichi Kikuta, Yoichi Takenaka, Masaru Ishii, Hideo Matsuda

  2014年10月, 英語


• A Segmentation Method for Bone Marrow Cavity Image Using Graph-Cuts

  Tomohiro Mashita, Jun Usami, Hironori Shigeta, Yoshihiro Kuroda, Junichi Kikuta, Shigeto Senoo, Masaru Ishii, Hideo Matsuda, Haruo Takemura

  2014年09月, 英語

  [査読有り]

  研究論文（国際会議プロシーディングス）


• A Graph Cuts Image Segmentation Method for Quantifying Barrier Permeation in Bone Tissue

  Hironori Shigeta, Tomohiro Mashita, Takeshi Kaneko, Junichi Kikuta, Shigeto Senoo, Haruo Takemura, Hideo Matsuda, Masaru Ishii

  2014年09月, 英語

  [査読有り]

  研究論文（国際会議プロシーディングス）


• Interferon-α acts on the S/G2/M phases to induce apoptosis in the G1 phase of an IFNAR2-expressing hepatocellular carcinoma cell line.

  Maeda S, Wada H, Naito Y, Nagano H, Simmons S, Kagawa Y, Naito A, Kikuta J, Ishii T, Tomimaru Y, Hama N, Kawamoto K, Kobayashi S, Eguchi H, Umeshita K, Ishii H, Doki Y, Mori M, Ishii M

  Interferon-α (IFN-α) is used clinically to treat hepatocellular carcinoma (HCC), although the detailed therapeutic mechanisms remain elusive. In particular, IFN-α has long been implicated in control of the cell cycle, but its actual point of action has not been clarified. Here, using time lapse imaging analyses of the human HCC cell line HuH7 carrying a fluorescence ubiquitination-based cell cycle indicator (Fucci), we found that IFN-α induced cell cycle arrest in the G0/G1 phases, leading to apoptosis through an IFN-α type-2 receptor (IFNAR2)-dependent signaling pathway. Detailed analyses by time lapse imaging and biochemical assays demonstrated that the IFN-α/IFNAR2 axis sensitizes cells to apoptosis in the S/G2/M phases in preparation for cell death in the G0/G1 phases. In summary, this study is the first to demonstrate the detailed mechanism of IFN-α as an anticancer drug, using Fucci-based time lapse imaging, which will be informative for treating HCC with IFN-α in clinical practice.

  2014年08月, The Journal of biological chemistry, 289(34) (34), 23786 - 95, 英語, 国際誌

  [査読有り]

  研究論文（学術雑誌）


• A Segmentation Method for Bone Marrow Cavity Imaging Using Graph Cuts

  Tomohiro Mashita, Jun Usam, Hironori Shigeta, Yoshihiro Kuroda, Junichi Kikuta, Shigeto Senoo, Masaru Ishi, Hideo Matsuda, Haruo Takemura

  IEEE, 2014年08月, 2014 1st Workshop on Pattern Recognition Techniques for Indirect Immunofluorescence Images

  研究論文（国際会議プロシーディングス）


• 生体二光子励起イメージングによる骨芽細胞・破骨細胞カップリングの可視化

  古家 雅之, 菊田 順一, 白崎 舞, 柏井 将文, 牧野 孝洋, 海渡 貴司, 岩崎 幹季, 石井 優, 吉川 秀樹

  (公社)日本整形外科学会, 2014年08月, 日本整形外科学会雑誌, 88(8) (8), S1416 - S1416, 日本語


• Left-right asymmetry is formed in individual cells by intrinsic cell chirality.

  Ryo Hatori, Tadashi Ando, Takeshi Sasamura, Naotaka Nakazawa, Mitsutoshi Nakamura, Kiichiro Taniguchi, Shunya Hozumi, Junichi Kikuta, Masaru Ishii, Kenji Matsuno

  2014年08月, Mechanisms of development, 133, 146 - 62, 英語, 国際誌

  [査読有り]

  研究論文（学術雑誌）


• Filamin acts as a key regulator in epithelial defence against transformed cells.

  Mihoko Kajita, Kaoru Sugimura, Atsuko Ohoka, Jemima Burden, Hitomi Suganuma, Masaya Ikegawa, Takashi Shimada, Tetsuya Kitamura, Masanobu Shindoh, Susumu Ishikawa, Sayaka Yamamoto, Sayaka Saitoh, Yuta Yako, Ryosuke Takahashi, Takaharu Okajima, Junichi Kikuta, Yumiko Maijima, Masaru Ishii, Masazumi Tada, Yasuyuki Fujita

  2014年07月, Nature communications, 5, 4428 - 4428, 英語, 国際誌

  [査読有り]

  研究論文（学術雑誌）


• [Bone metabolism and cardiovascular function update. Relationship between immune and vascular system in bone tissues].

  Junichi Kikuta, Masaru Ishii

  Bone marrow is a highly vascularized organ and the vessel walls have a large number of fenestrations. Various kinds of hematopoietic cells are continuously entering into and egressing from bone marrow cavity through these unique features. Monocytoid osteoclast precursors also migrate across the capillary endothelium, serving as a dynamic point of control of bone homeostasis. We have recently revealed the cellular dynamics of monocytoid osteoclast precursors in vivo by a novel fluorescent imaging technology. Furthermore we found the regulatory mechanism of vascular permeability in bone marrow in vivo by using intravital multiphoton microscopy. Here we show the latest data and also discuss its further application.

  2014年07月, Clinical calcium, 24(7) (7), 63 - 8, 日本語, 国内誌

  [査読有り]

  研究論文（学術雑誌）


• 脂肪組織慢性炎症における免疫細胞の動態変化の解析

  堰本 亮平, 前田 法一, 對馬 佑, 松田 圭介, 森 卓也, 福田 士郎, 中辻 秀朗, 西澤 均, 岸田 堅, 菊田 順一, 船橋 徹, 石井 優, 下村 伊一郎

  (一社)日本動脈硬化学会, 2014年06月, 日本動脈硬化学会総会プログラム・抄録集, 46回, 320 - 320, 日本語


• Olopatadine hydrochloride restores histamine-induced impaired sweating.

  Saki Matsui, Hiroyuki Murota, Emi Ono, Junichi Kikuta, Masaru Ishii, Ichiro Katayama

  2014年06月, Journal of dermatological science, 74(3) (3), 260 - 1, 英語, 国際誌

  [査読有り]


• Generation of colonic IgA-secreting cells in the caecal patch.

  Kazunori Masahata, Eiji Umemoto, Hisako Kayama, Manato Kotani, Shota Nakamura, Takashi Kurakawa, Junichi Kikuta, Kazuyoshi Gotoh, Daisuke Motooka, Shintaro Sato, Tomonori Higuchi, Yoshihiro Baba, Tomohiro Kurosaki, Makoto Kinoshita, Yosuke Shimada, Taishi Kimura, Ryu Okumura, Akira Takeda, Masaru Tajima, Osamu Yoshie, Masahiro Fukuzawa, Hiroshi Kiyono, Sidonia Fagarasan, Tetsuya Iida, Masaru Ishii, Kiyoshi Takeda

  2014年04月, Nature communications, 5, 3704 - 3704, 英語, 国際誌

  [査読有り]

  研究論文（学術雑誌）


• [Bone and Stem Cells. Intravital imaging of bone marrow microenvironment].

  Hiroki Mizuno, Junichi Kikuta, Masaru Ishii

  Various kinds of cell types, such as osteoclasts, osteoblasts, hematopoietic cells, and mesenchymal cells, have been reported to exist in the bone marrow and communicate with each other. Although there have been many previous studies about bone marrow microenvironment, most of them were analyzed by conventional methods such as histological analysis and flow cytometry. These methods could not observe the dynamic cell movement in living bone marrow. Recently rapid development of fluorescent imaging techniques enables us to understand the cellular dynamics in vivo . That's why we have originally established an advanced imaging system for visualizing living bone tissues with intravital two-photon microscopy. Here we show the latest data and the detailed methodology of intravital imaging of bone marrow microenvironment, and also discuss its further application.

  2014年04月, Clinical calcium, 24(4) (4), 541 - 6, 日本語, 国内誌

  [査読有り]

  研究論文（学術雑誌）


• Lysophosphatidic acid receptor type 1 (LPA1) plays a functional role in osteoclast differentiation and bone resorption activity.

  Marion David, Irma Machuca-Gayet, Junichi Kikuta, Penelope Ottewell, Fuka Mima, Raphael Leblanc, Edith Bonnelye, Johnny Ribeiro, Ingunn Holen, Rùben Lopez Vales, Pierre Jurdic, Jerold Chun, Philippe Clézardin, Masaru Ishii, Olivier Peyruchaud

  2014年03月, The Journal of biological chemistry, 289(10) (10), 6551 - 64, 英語, 国際誌

  [査読有り]

  研究論文（学術雑誌）


• Dynamic analysis of histamine-mediated attenuation of acetylcholine-induced sweating via GSK3β activation.

  Matsui S, Murota H, Takahashi A, Yang L, Lee JB, Omiya K, Ohmi M, Kikuta J, Ishii M, Katayama I

  2014年02月, The Journal of investigative dermatology, 134(2) (2), 326 - 334, 英語, 国際誌

  [査読有り]

  研究論文（学術雑誌）


• Intravital multiphoton microscopy for dissecting cellular dynamics in arthritic inflammation and bone destruction.

  Junichi Kikuta, Masaru Ishii

  Osteoclasts are giant bone-resorbing polykaryons that differentiate from mononuclear macrophage/monocyte-lineage hematopoietic precursors. They play critical roles not only in normal bone homeostasis (remodeling) but also in the pathogenesis of bone-destructive disorders such as osteoporosis and rheumatoid arthritis. However, how the activity of mature osteoclasts is regulated in vivo remains unclear. To answer this question, we recently developed an advanced imaging system to visualize living bone tissues with intravital multiphoton microscopy. Using this system, we succeeded in visualization of mature osteoclasts in living bones. We herein describe the detailed methodology for visualizing bone resorption of mature osteoclasts in living bone marrow and joints using intravital multiphoton microscopy. This approach would be beneficial for studying the cellular dynamics in arthritic inflammation and bone destruction in vivo and would thus be useful for evaluating novel anti-bone-resorptive drugs.

  2014年, Methods in molecular biology (Clifton, N.J.), 1142, 1 - 10, 英語, 国際誌

  [査読有り]

  研究論文（学術雑誌）


• [Dynamics of bone resorption analyzed by intravital imaging].

  Junichi Kikuta, Masaru Ishii

  By using conventional methods such as histological analysis, many osteoclasts can be observed in the site of bone destruction. However, how the bone-resorptive functions of mature osteoclasts are controlled in vivo remains unclear. To answer this question, we have originally developed an advanced imaging system for visualizing living bone tissues with intravital multiphoton microscopy. Using this system, we succeeded in visualizing bone resorption of mature osteoclasts in living bone. Here we show the latest data and also discuss the further application of intravital bone imaging. This approach would be quite useful for evaluating novel anti-bone-resorptive drugs in vivo .

  2013年11月, Clinical calcium, 23(11) (11), 1627 - 33, 日本語, 国内誌

  [査読有り]

  研究論文（学術雑誌）


• Sphingosine-1-phosphate and chemokines in the control of osteoclast precursors migration visualized by intravital multiphoton microscopy

  Junichi Kikuta, Atsuko Kubo, Masaru Ishii

  2013年10月, Osteoimmunology: Interactions of the immune and skeletal systems, 73 - 79

  [査読有り]

  論文集(書籍)内論文


• Dynamic live imaging of bone: opening a new era with 'bone histodynametry'.

  Masaru Ishii, Sayumi Fujimori, Takeshi Kaneko, Junichi Kikuta

  Recent advances in optical imaging with two-photon excitation microscopy have enabled visualization of the inside of intact bone tissues in living animals. Using these advanced techniques, the dynamic behaviors of live bone cells and static histological information on bone tissue structures can be elucidated. The migration and positioning of osteoclast precursor monocytes, the bone-resorbing function of mature osteoclasts, and its functional coupling with bone-replenishing osteoblasts have been evaluated, including their dynamic properties in intact live bones. This novel 'bone histodynametric' methodology, combined with conventional histomorphometric analyses, will surely contribute to opening of a new era in bone and mineral research.

  2013年09月, Journal of bone and mineral metabolism, 31(5) (5), 507 - 11, 英語, 国内誌

  [査読有り]

  研究論文（学術雑誌）


• Layer V cortical neurons require microglial support for survival during postnatal development.

  Masaki Ueno, Yuki Fujita, Tatsuhide Tanaka, Yuka Nakamura, Junichi Kikuta, Masaru Ishii, Toshihide Yamashita

  Neurons require trophic support during neural circuit formation; however, how the cellular milieu contributes to neuronal survival remains unclear. We found that layer V cortical neurons require support from microglia for survival during postnatal development. Specifically, we found that microglia accumulated close to the subcerebral and callosal projection axons in the postnatal brain. Inactivation of microglia by minocycline treatment or transient ablation of microglia in CD11b-DTR transgenic mice led to increased apoptosis, specifically in layer V subcerebral and callosal projection neurons. CX3CR1 in microglia was required for the survival of layer V neurons. Microglia consistently promoted the survival of cortical neurons in vitro. In addition, we identified microglia-derived IGF1 as a trophic factor that maintained neuronal survival. Our results highlight a neuron-glia interaction that is indispensable for network formation during a specific period in the developing brain.

  2013年05月, Nature neuroscience, 16(5) (5), 543 - 51, 英語, 国際誌

  [査読有り]

  研究論文（学術雑誌）


• Sphingosine-1-phosphate-mediated osteoclast precursor monocyte migration is a critical point of control in antibone-resorptive action of active vitamin D.

  Junichi Kikuta, Shunsuke Kawamura, Fumie Okiji, Mai Shirazaki, Sadaoki Sakai, Hitoshi Saito, Masaru Ishii

  National Academy of Sciences, 2013年04月, Proceedings of the National Academy of Sciences of the United States of America, 110(17) (17), 7009 - 13, 英語, 国際誌

  [査読有り]

  研究論文（学術雑誌）


• [Analysis of bone tissues by using fluorescent imaging].

  Junichi Kikuta, Masaru Ishii

  Rapid development of fluorescent imaging techniques enables us to understand cellular dynamics in vivo . We have originally established an advanced imaging system for visualizing live bone tissues with intravital multiphoton microscopy. By means of the system we have recently succeeded in visualization of mature osteoclasts in live bones and revealed that RANKL regulates bone-resorptive functions of mature osteoclasts in vivo . We also developed new image analysis software for tracking morphological changes of mature osteoclasts and generated pH-sensing chemical fluorescent probes for detecting bone resorption at local sites on the bone surface. Here we show the latest data and the detailed methodology of intravital imaging of bone tissues, and also discuss its further application.

  2013年03月, Clinical calcium, 23(3) (3), 355 - 60, 日本語, 国内誌

  [査読有り]

  研究論文（学術雑誌）


• Osteoclast migration, differentiation and function: novel therapeutic targets for rheumatic diseases.

  Junichi Kikuta, Masaru Ishii

  RA is a chronic autoimmune disease characterized by joint synovial inflammation and progressive cartilage/bone destruction. Although various kinds of RA drug have been developed worldwide, there are currently no established methods for preventing RA-associated bone destruction, the most severe outcome of this disease. One of the major pathogenic factors in arthritic bone destruction is the enhanced activity of osteoclasts at inflammatory sites. Osteoclasts are bone-resorbing giant polykaryons that differentiate from mononuclear macrophage/monocyte-lineage haematopoietic precursors. Upon stimulation by cytokines, such as M-CSF and RANK ligand, osteoclast precursor monocytes migrate and attach onto the bone surface (migration). They then fuse with each other to form giant cells (differentiation) and mediate bone resorption (function). In this review, we summarize the current understanding regarding the mechanisms underlying these three dynamic steps of osteoclastic activity and discuss novel lines of osteoclast-targeted therapies that will impact future treatment of RA.

  2013年02月, Rheumatology (Oxford, England), 52(2) (2), 226 - 34, 英語, 国際誌

  [査読有り]

  研究論文（学術雑誌）


• Dynamic visualization of RANKL and Th17-mediated osteoclast function.

  Junichi Kikuta, Yoh Wada, Toshiyuki Kowada, Ze Wang, Ge-Hong Sun-Wada, Issei Nishiyama, Shin Mizukami, Nobuhiko Maiya, Hisataka Yasuda, Atsushi Kumanogoh, Kazuya Kikuchi, Ronald N Germain, Masaru Ishii

  Osteoclasts are bone resorbing, multinucleate cells that differentiate from mononuclear macrophage/monocyte-lineage hematopoietic precursor cells. Although previous studies have revealed important molecular signals, how the bone resorptive functions of such cells are controlled in vivo remains less well characterized. Here, we visualized fluorescently labeled mature osteoclasts in intact mouse bone tissues using intravital multiphoton microscopy. Within this mature population, we observed cells with distinct motility behaviors and function, with the relative proportion of static - bone resorptive (R) to moving - nonresorptive (N) varying in accordance with the pathophysiological conditions of the bone. We also found that rapid application of the osteoclast-activation factor RANKL converted many N osteoclasts to R, suggesting a novel point of action in RANKL-mediated control of mature osteoclast function. Furthermore, we showed that Th17 cells, a subset of RANKL-expressing CD4+ T cells, could induce rapid N-to-R conversion of mature osteoclasts via cell-cell contact. These findings provide new insights into the activities of mature osteoclasts in situ and identify actions of RANKL-expressing Th17 cells in inflammatory bone destruction.

  2013年02月, The Journal of clinical investigation, 123(2) (2), 866 - 73, 英語, 国際誌

  [査読有り]

  研究論文（学術雑誌）


• Systemic circulation and bone recruitment of osteoclast precursors tracked by using fluorescent imaging techniques.

  Manato Kotani, Junichi Kikuta, Frederick Klauschen, Takenao Chino, Yasuhiro Kobayashi, Hisataka Yasuda, Katsuto Tamai, Atsushi Miyawaki, Osami Kanagawa, Michio Tomura, Masaru Ishii

  2013年01月, Journal of immunology (Baltimore, Md. : 1950), 190(2) (2), 605 - 12, 英語, 国際誌

  [査読有り]

  研究論文（学術雑誌）


• Sphingosine-1-phosphate signaling controlling osteoclasts and bone homeostasis.

  Masaru Ishii, Junichi Kikuta

  Bone is a dynamic organ that is continuously turned over during growth, even in adults. During bone remodeling, homeostasis is regulated by the balance between bone formation by osteoblasts and bone resorption by osteoclasts. However, in pathological conditions such as osteoporosis, osteopetrosis, arthritic joint destruction, and bone metastasis, this equilibrium is disrupted. Since osteoclasts are excessively activated in osteolytic diseases, the inhibition of osteoclast function has been a major therapeutic strategy. It has recently been demonstrated that sphingosine-1-phosphate (S1P), a biologically active lysophospholipid that is enriched in blood, controls the trafficking of osteoclast precursors between the circulation and bone marrow cavities via G protein-coupled receptors, S1PRs. While S1PR1 mediates chemoattraction toward S1P in bone marrow, where S1P concentration is low, S1PR2 mediates chemorepulsion in blood, where the S1P concentration is high. The regulation of precursor recruitment may represent a novel therapeutic strategy for controlling osteoclast-dependent bone remodeling. By means of intravital multiphoton imaging of bone tissues, we have recently revealed that the reciprocal action of S1P controls the migration of osteoclast precursors between bone tissues and blood stream. Imaging technologies have enabled us to visualize the in situ behaviors of different cell types in intact tissues. In this review we also discuss future perspectives on this new method in the field of bone biology and medical sciences in general. This article is part of a Special Issue entitled Advances in Lysophospholipid Research.

  2013年01月, Biochimica et biophysica acta, 1831(1) (1), 223 - 7, 英語, 国際誌

  [査読有り]

  研究論文（学術雑誌）


• Cell cycle-dependent Rho GTPase activity dynamically regulates cancer cell motility and invasion in vivo.

  Yoshinori Kagawa, Shinji Matsumoto, Yuji Kamioka, Koshi Mimori, Yoko Naito, Taeko Ishii, Daisuke Okuzaki, Naohiro Nishida, Sakae Maeda, Atsushi Naito, Junichi Kikuta, Keizo Nishikawa, Junichi Nishimura, Naotsugu Haraguchi, Ichiro Takemasa, Tsunekazu Mizushima, Masataka Ikeda, Hirofumi Yamamoto, Mitsugu Sekimoto, Hideshi Ishii, Yuichiro Doki, Michiyuki Matsuda, Akira Kikuchi, Masaki Mori, Masaru Ishii

  The mechanism behind the spatiotemporal control of cancer cell dynamics and its possible association with cell proliferation has not been well established. By exploiting the intravital imaging technique, we found that cancer cell motility and invasive properties were closely associated with the cell cycle. In vivo inoculation of human colon cancer cells bearing fluorescence ubiquitination-based cell cycle indicator (Fucci) demonstrated an unexpected phenomenon: S/G2/M cells were more motile and invasive than G1 cells. Microarray analyses showed that Arhgap11a, an uncharacterized Rho GTPase-activating protein (RhoGAP), was expressed in a cell-cycle-dependent fashion. Expression of ARHGAP11A in cancer cells suppressed RhoA-dependent mechanisms, such as stress fiber formation and focal adhesion, which made the cells more prone to migrate. We also demonstrated that RhoA suppression by ARHGAP11A induced augmentation of relative Rac1 activity, leading to an increase in the invasive properties. RNAi-based inhibition of Arhgap11a reduced the invasion and in vivo expansion of cancers. Additionally, analysis of human specimens showed the significant up-regulation of Arhgap11a in colon cancers, which was correlated with clinical invasion status. The present study suggests that ARHGAP11A, a cell cycle-dependent RhoGAP, is a critical regulator of cancer cell mobility and is thus a promising therapeutic target in invasive cancers.

  2013年, PloS one, 8(12) (12), e83629, 英語, 国際誌

  [査読有り]

  研究論文（学術雑誌）


• Use of intravital microscopy and in vitro chemotaxis assays to study the roles of sphingosine-1-phosphate in bone homeostasis.

  Taeko Ishii, Shunsuke Kawamura, Issei Nishiyama, Junichi Kikuta, Masaru Ishii

  We describe a method to visualize the migration of osteoclast precursors within intact murine bone -marrow in real time using intravital multiphoton microscopy. Conventionally, cell migration has been evaluated using in vitro systems, such as transmigration assays. Although these methods are convenient for quantification and are highly reproducible, these in vitro assay systems may not accurately reflect in vivo cellular behavior. In addition to in vitro analyses, recent technological progress in two-photon excitation-based laser microscopy has enabled the visualization of dynamic cell behavior deep inside intact living organs. Combining this imaging method with in vitro chemoattraction analyses, we have revealed that sphingosine-1-phosphate (S1P), a lipid mediator enriched in blood, bidirectionally controls the trafficking of osteoclast precursors between the circulation and bone marrow cavities via G protein-coupled receptors (GPCRs).

  2012年, Methods in molecular biology (Clifton, N.J.), 874, 129 - 39, 英語, 国際誌

  [査読有り]

  研究論文（学術雑誌）


• Recent advances in intravital imaging of dynamic biological systems.

  Junichi Kikuta, Masaru Ishii

  Intravital multiphoton microscopy has opened a new era in the field of biological imaging. Focal excitation of fluorophores by simultaneous attack of multiple (normally "two") photons generates images with high spatial resolution, and use of near-infrared lasers for multiphoton excitation allows penetration of thicker specimens, enabling biologists to visualize living cellular dynamics deep inside tissues and organs without thin sectioning. Moreover, the minimized photo-bleaching and toxicity associated with multiphoton techniques is beneficial for imaging of live specimens for extended observation periods. Here we focus on recent findings using intravital multiphoton imaging of dynamic biological systems such as the immune system and bone homeostasis. The immune system comprises highly dynamic networks, in which many cell types actively travel throughout the body and interact with each other in specific areas. Therefore, real-time intravital imaging represents a powerful tool for understanding the mechanisms underlying this dynamic system.

  2012年, Journal of pharmacological sciences, 119(3) (3), 193 - 7, 英語, 国内誌

  [査読有り]

  研究論文（学術雑誌）


• [Bone and calcium update; bone research update. Trafficking of osteoclast precursors visualized by intravital imaging technology].

  Manato Kotani, Junichi Kikuta, Masaru Ishii

  Osteoclasts are 'bone-resorbing' giant polykaryons that differentiate from mononuclear macrophage/monocyte-lineage hematopoietic precursors. However, it has remained unclear how osteoclast precursors migrate into the bone surface and what controls their migratory behaviors in vivo . To investigate these questions, we utilized an advanced imaging system for visualizing live bone tissues with intravital multiphoton microscopy that we have recently established. By means of the system we have recently succeeded in visualization of osteoclast precursors in live bones and revealed that sphingosine-1-phosphate (S1P) , a lipid mediator, dynamically regulates migration and localization of osteoclasts and their precursors in vivo . Here we show the latest data and the detailed methodology of intravital imaging of bone tissues, and also discuss its further application.

  2011年12月, Clinical calcium, 21(12) (12), 85 - 91, 日本語, 国内誌

  [査読有り]

  研究論文（学術雑誌）


• In vivo fluorescence imaging of bone-resorbing osteoclasts.

  Toshiyuki Kowada, Junichi Kikuta, Atsuko Kubo, Masaru Ishii, Hiroki Maeda, Shin Mizukami, Kazuya Kikuchi

  2011年11月, Journal of the American Chemical Society, 133(44) (44), 17772 - 6, 英語, 国際誌

  [査読有り]

  研究論文（学術雑誌）


• 【生きたままの姿を見る4Dイメージング-免疫・癌・脳神経の時空間的ダイナミクス】免疫・血液系の2光子励起イメージング"soft-wired network"の実体的解明

  菊田 順一, 久保 厚子, 島津 裕, 石井 優

  (株)羊土社, 2011年10月, 実験医学, 29(16) (16), 2602 - 2606, 日本語

  [査読有り]


• [In vivo imaging of mature osteoclasts and RANKL signaling].

  Junichi Kikuta, Masaru Ishii

  Osteoclasts are 'bone-resorbing' giant polykaryons that differentiate from mononuclear macrophage/monocyte-lineage hematopoietic precursors. However how the activity of mature osteoclasts is regulated in vivo remains unclear. To answer the question, we utilized an advanced imaging system for visualizing live bone tissues with intravital multiphoton microscopy that we have recently established. By means of the system we have recently succeeded in visualization of mature osteoclasts in live bones and revealed that RANKL regulates bone-resorptive functions of mature osteoclasts in vivo . Here we show the latest data and the detailed methodology of intravital imaging of bone tissues, and also discuss its further application.

  2011年08月, Clinical calcium, 21(8) (8), 1181 - 5, 日本語, 国内誌

  [査読有り]

  研究論文（学術雑誌）


• S1P-targeted therapy for elderly rheumatoid arthritis patients with osteoporosis.

  Junichi Kikuta, Kaori Iwai, Yukihiko Saeki, Masaru Ishii

  2011年07月, Rheumatology international, 31(7) (7), 967 - 9, 英語, 国際誌

  [査読有り]

  研究論文（学術雑誌）


• ヒト破骨細胞の遊走を標的とした新規骨粗鬆症治療薬の開発と応用

  石井 優, 菊田 順一, 島津 裕, 久保 厚子, 岩井 香織, 佐伯 行彦

  (公財)臨床薬理研究振興財団, 2011年06月, 臨床薬理の進歩, (32) (32), 11 - 17, 日本語

  [査読有り]


• The role of sphingosine 1-phosphate in migration of osteoclast precursors; an application of intravital two-photon microscopy.

  Taeko Ishii, Yutaka Shimazu, Issei Nishiyama, Junichi Kikuta, Masaru Ishii

  Sphingosine-1-phosphate (S1P), a biologically active lysophospholipid that is enriched in blood, controls the trafficking of osteoclast precursors between the circulation and bone marrow cavities via G protein-coupled receptors, S1PRs. While S1PR1 mediates chemoattraction toward S1P in bone marrow, where S1P concentration is low, S1PR2 mediates chemorepulsion in blood, where the S1P concentration is high. The regulation of precursor recruitment may represent a novel therapeutic strategy for controlling osteoclast-dependent bone remodeling. Through intravital multiphoton imaging of bone tissues, we reveal that the bidirectional function of S1P temporospatially regulates the migration of osteoclast precursors within intact bone tissues. Imaging technologies have enabled in situ visualization of the behaviors of several players in intact tissues. In addition, intravital microscopy has the potential to be more widely applied to functional analysis and intervention.

  2011年05月, Molecules and cells, 31(5) (5), 399 - 403, 英語, 国際誌

  [査読有り]

  研究論文（学術雑誌）


• [Encounter of cancer cells with bone. In vivo imaging of osteoclasts and their precursors in intact bone tissues].

  Junichi Kikuta, Shunsuke Kawamura, Masaru Ishii

  Osteoclasts play critical roles not only in normal bone homeostasis ('remodeling') , but also in the pathogenesis of bone destructive disorders such as osteoporosis, rheumatoid arthritis, and bone metastasis. However, it has not been known how osteoclast precursor monocytes migrate into the bone surface and what controls their migratory behaviors. To reveal these systems, we have recently established a new system for visualizing intact bone tissues and bone marrow cavities in live animals by using an advanced imaging technique with intravital two-photon microscopy. By means of the system we have revealed that sphingosine-1-phosphate (S1P) , a lipid mediator, dynamically regulates migration and localization of osteoclasts and their precursors in vivo . Here we show the latest data and the detailed methodology of intravital imaging of bone tissues, and also discuss its further application.

  2011年03月, Clinical calcium, 21(3) (3), 372 - 8, 日本語, 国内誌

  [査読有り]

  研究論文（学術雑誌）


• Chemorepulsion by blood S1P regulates osteoclast precursor mobilization and bone remodeling in vivo.

  Masaru Ishii, Junichi Kikuta, Yutaka Shimazu, Martin Meier-Schellersheim, Ronald N Germain

  Sphingosine-1-phosphate (S1P), a lipid mediator enriched in blood, controls the dynamic migration of osteoclast (OC) precursors (OPs) between the blood and bone, in part via the S1P receptor 1 (S1PR1) which directs positive chemotaxis toward S1P. We show that OPs also express S1PR2, an S1P receptor which mediates negative chemotaxis (or chemorepulsion). OP-positive chemotaxis is prominent in gradients with low maximal concentrations of S1P, whereas such behavior is minimal in fields with high maximal S1P concentrations. This reverse-directional behavior is caused by S1PR2-mediated chemorepulsion acting to override S1PR1 upgradient motion. S1PR2-deficient mice exhibit moderate osteopetrosis as a result of a decrease in osteoclastic bone resorption, suggesting that S1PR2 contributes to OP localization on the bones mediated by chemorepulsion away from the blood where S1P levels are high. Inhibition of S1PR2 function by the antagonist JTE013 changed the migratory behavior of monocytoid cells, including OPs, and relieved osteoporosis in a mouse model by limiting OP localization and reducing the number of mature OCs attached to the bone surface. Thus, reciprocal regulation of S1P-dependent chemotaxis controls bone remodeling by finely regulating OP localization. This regulatory axis may be promising as a therapeutic target in diseases affecting OC-dependent bone remodeling.

  2010年12月, The Journal of experimental medicine, 207(13) (13), 2793 - 8, 英語, 国際誌

  [査読有り]

  研究論文（学術雑誌）


• 自己免疫疾患・リウマチ性疾患・免疫不全症 骨髄線維症を認めた混合性結合組織病の一症例

  山中 隆夫, 原田 芳徳, 石井 泰子, 菊田 順一, 田中 枝里子, 石井 優, 松下 正人, 大島 至郎, 佐伯 行彦, 片田 圭宣

  (一社)日本アレルギー学会, 2009年04月, アレルギー, 58(3-4) (3-4), 380 - 380, 日本語


• Carvedilol blocks cardiac K-ATP and K-G but not I-K1 channels by acting at the bundle-crossing regions

  J Kikuta, M Ishii, K Kishimoto, Y Kurachi

  2006年01月, EUROPEAN JOURNAL OF PHARMACOLOGY, 529(1-3) (1-3), 47 - 54, 英語

  [査読有り]

  研究論文（学術雑誌）

• 歯周炎による歯槽骨破壊を引き起こす破骨前駆細胞の由来

  纐纈 友斗, 岩山 智明, 阪下 裕美, 揚村 朋弥, 菊田 順一, 石井 優, 吉田 悠作, 松本 修治, Bhongsatiern Phan, 坪井 栄生, 村上 伸也, 竹立 匡秀

  (NPO)日本歯周病学会, 2024年10月, 日本歯周病学会会誌, 66(秋季特別) (秋季特別), 140 - 140, 日本語


• 組織線維化と線維症 組織線維化と線維症 その生理と病理

  田所 慶誠, 菊田 順一, 石井 優

  (株)先端医学社, 2024年08月, 炎症と免疫, 32(5) (5), 425 - 430, 日本語


• 生体イメージングで捉える骨代謝ダイナミクス

  菊田 順一

  (一社)日本内分泌学会, 2024年05月, 日本内分泌学会雑誌, 100(1) (1), 265 - 265, 日本語


• 歯周組織由来破骨前駆細胞の解析

  阪下 裕美, 岩山 智明, 松本 修治, 岩下 瑞穂, Bhongsatiern Phan, 纐纈 友斗, 吉田 悠作, 菊田 順一, 上中 麻希, 鎗 信弥, 揚村 朋弥, 石井 優, 村上 伸也

  (NPO)日本歯周病学会, 2023年10月, 日本歯周病学会会誌, 65(秋季特別) (秋季特別), 130 - 130, 日本語


• 【膠原病】炎症性骨破壊における破骨前駆細胞

  揚村 朋弥, 菊田 順一, 石井 優

  (株)北隆館, 2023年10月, 別冊Bio Clinica: 慢性炎症と疾患, 12(2) (2), 54 - 58, 日本語


• 関節リウマチ・膠原病診療のピットフォールを回避するために(第4回)(No.7) 骨粗鬆症 関節リウマチ患者における骨粗鬆症のマネジメントについて

  金子 雄, 菊田 順一, 石井 優, 山岡 邦宏

  (株)先端医学社, 2023年08月, 炎症と免疫, 31(5) (5), 461 - 465, 日本語


• 骨・免疫系の生体イメージング研究—Intravital imaging of bone and immune system—特集 生体イメージングの最前線 : 絶え間ない技術革新と生命医科学の新展開 ; イメージングで免疫系や生体恒常性を解析する

  佐藤 和真, 鎗 伸弥, 菊田 順一

  医歯薬出版, 2023年07月29日, 医学のあゆみ, 286(5) (5), 392 - 398, 日本語


• 生体骨イメージングによる骨芽前駆細胞の動態解析

  森本 彬人, 執行 穂高, 菊田 順一, 石井 優

  (一社)日本骨代謝学会, 2023年07月, 日本骨代謝学会学術集会プログラム抄録集, 41回, 127 - 127, 日本語


• シングルセルRNAシーケンスによる関節炎特異的な破骨前駆細胞の分化経路解析

  揚村 朋弥, 菊田 順一, 石井 優

  (一社)日本骨代謝学会, 2023年07月, 日本骨代謝学会学術集会プログラム抄録集, 41回, 141 - 141, 日本語


• 光-電子相関顕微鏡法(CLEM)とFocused Ion Beam Scanning Electron Microscope(FIB-SEM)を用いた破骨細胞の三次元構造解析

  細沼 雅弘, 坂井 信裕, 松島 英輝, 菊田 順一, 高木 孝士, 武部 明, 石井 優, 高見 正道

  (一社)日本骨代謝学会, 2023年07月, 日本骨代謝学会学術集会プログラム抄録集, 41回, 143 - 143, 日本語


• RANKLによる生体骨髄内の血管透過性制御機構の解明

  菊田 順一, 石井 優

  (一社)日本骨代謝学会, 2023年07月, 日本骨代謝学会学術集会プログラム抄録集, 41回, 153 - 153, 日本語


• 【ビタミンと免疫】マクロファージ分化におけるビタミンDの作用

  佐藤 和真, 粟生 智香, 菊田 順一, 石井 優

  (有)科学評論社, 2023年06月, 臨床免疫・アレルギー科, 79(6) (6), 631 - 636, 日本語


• 光-電子相関顕微鏡法とFocused Ion Beam Scanning Electron Microscopeを用いた破骨細胞の三次元構造解析

  細沼 雅弘, 坂井 信裕, 松島 英輝, 菊田 順一, 高木 孝士, 武部 明, 石井 優, 高見 正道

  日本骨形態計測学会, 2023年05月, 日本骨形態計測学会雑誌, 33(1) (1), 198 - 198, 日本語


• 骨組織pHを測定するレシオ蛍光イメージングプローブの開発

  三木 初音, 吉村 康孝, 蓑島 維文, 鎗 伸弥, 菊田 順一, 石井 優, 菊地 和也

  日本分子イメージング学会, 2023年05月, JSMI Report, 16(2) (2), 110 - 110, 日本語


• 生体イメージングによる炎症性破骨細胞の動態・機能解析

  菊田 順一, 石井 優

  (一社)日本リウマチ学会, 2023年03月, 日本リウマチ学会総会・学術集会プログラム・抄録集, 67回, 766 - 766, 日本語


• 骨組織の生体イメージング—Intravital imaging of bone tissues—特集 新組織学シリーズ(4)骨・軟骨 ; 骨・軟骨組織の構造と細胞分化・機能

  鎗 伸弥, 菊田 順一, 石井 優

  金原一郎記念医学医療振興財団, 2023年, 生体の科学, 74(6) (6), 540 - 543, 日本語


• 【最新の骨粗鬆症学(第2版)-骨粗鬆症学の最新知見-】骨研究フロンティア 骨の構造と機能 骨組織の動態機構とその制御

  上中 麻希, 菊田 順一, 石井 優

  (株)日本臨床社, 2023年01月, 日本臨床, 81(増刊1 最新の骨粗鬆症学) (増刊1 最新の骨粗鬆症学), 64 - 70, 日本語


• Arthritis-associated osteoclastogenic macrophage, AtoM, as a key player in pathological bone erosion

  Tomoya Agemura, Tetsuo Hasegawa, Shinya Yari, Junichi Kikuta, Masaru Ishii

  2022年12月, Inflammation and Regeneration, 42(1) (1)

  書評論文,書評,文献紹介等


• 臨床免疫領域における画像・組織による診断・病型分類の役割 生体多光子励起イメージングによる免疫疾患の病態解明

  菊田 順一, 石井 優

  (一社)日本臨床免疫学会, 2022年10月, 日本臨床免疫学会総会プログラム・抄録集, 50回, 60 - 60, 日本語


• 【シングルセル解析による免疫細胞研究の新展開】生体イメージングとオミックス解析の統合による炎症性病態の真相解明

  宮本 佑, 菊田 順一, 石井 優

  (株)先端医学社, 2022年10月, 炎症と免疫, 30(6) (6), 495 - 500, 日本語


• Vocabulary 生体多光子励起イメージング

  鎗 伸弥, 菊田 順一, 石井 優

  (株)南江堂, 2022年10月, 整形外科, 73(11) (11), 1168 - 1168, 日本語


• 【自己免疫疾患 層別化する新時代へ 臨床検体のマルチオミクス解析、腸内細菌によって見えてきた免疫経路の全容】(第3章)自己免疫疾患の特異的病態経路の解明 生体イメージングが照らし出す臓器炎症プロセスの実態

  宮本 佑, 菊田 順一, 石井 優

  (株)羊土社, 2022年09月, 実験医学, 40(15) (15), 2539 - 2545, 日本語


• 【基盤病態としての慢性炎症】慢性炎症の生体イメージング

  揚村 朋弥, 菊田 順一, 石井 優

  医歯薬出版(株), 2022年07月, 医学のあゆみ, 282(1) (1), 105 - 111, 日本語


• Rising Stars in Skeletal Biology 骨芽細胞由来の細胞外小胞は骨形成期から骨吸収期への相転換に関与する

  上中 麻希, 菊田 順一, 石井 優

  (一社)日本骨代謝学会, 2022年07月, 日本骨代謝学会学術集会プログラム抄録集, 40回, 94 - 94, 日本語


• 二光子イメージングを用いた細胞外ATP依存的なCD137アゴニスト抗体による全身免疫活性化低減の検証

  金子 知里, 筒井 遥香, 尾関 和久, 本多 正樹, 原谷 健太, 成田 義規, 櫻井 実香, 菊田 順一, 田保 充康, 石井 優

  (一社)日本毒性学会, 2022年06月, The Journal of Toxicological Sciences, 47(Suppl.) (Suppl.), S110 - S110, 日本語


• 骨ミネラル代謝の新知見 SLPI PTH誘導性骨形成における新規メディエーターの発見

  石井 優, 菊田 順一

  (一社)日本腎臓学会, 2022年05月, 日本腎臓学会誌, 64(3) (3), 209 - 209, 日本語


• 【最新関節リウマチ学(第2版)-寛解・治癒を目指した研究と最新治療-】関節リウマチの臓器障害と合併症・薬剤副作用対策 骨粗鬆症、骨折(薬剤性を含む)

  塚崎 裕之, 菊田 順一, 石井 優

  (株)日本臨床社, 2022年04月, 日本臨床, 80(増刊4 最新関節リウマチ学) (増刊4 最新関節リウマチ学), 625 - 629, 日本語


• 【シン・マクロファージ あらゆる疾患を制御する機能的多様性】(第2章)病気とマクロファージの多様性を知る14の方法 関節破壊を惹起する炎症性マクロファージの同定 破骨前駆細胞の新たなサブセット

  揚村 朋弥, 長谷川 哲雄, 菊田 順一, 石井 優

  (株)羊土社, 2022年03月, 実験医学, 40(5) (5), 697 - 703, 日本語


• Arthritis-associated osteoclastogenic macrophages (AtoMs) participate in pathological bone erosion in rheumatoid arthritis

  Tomoya Agemura, Tetsuo Hasegawa, Shinya Yari, Junichi Kikuta, Masaru Ishii

  2022年, Immunological Medicine, 45(1) (1), 22 - 26

  書評論文,書評,文献紹介等


• 【夜明けを迎えたヒト免疫学 臨床検体からヒトの免疫応答機構の実態を知り、疾患制御に挑む】生体ライブイメージング技術から展開する新たなヒト免疫学

  宮本 佑, 菊田 順一, 石井 優

  (株)羊土社, 2022年01月, 実験医学, 40(1) (1), 47 - 53, 日本語


• 見る脂質のページ イメージングと細胞解析でみるイムノメタボリズム(第2回) In vivoライブイメージング技術を活用したイムノメタボリズムの見える化

  宮本 佑, 菊田 順一, 石井 優

  (株)メディカルレビュー社, 2021年10月, The Lipid, 32(2) (2), 106 - 113, 日本語


• 生体イメージングの進歩と新たな技術開発への挑戦

  粟生 智香, 菊田 順一, 石井 優

  (公社)日本薬学会, 2021年10月, ファルマシア, 57(10) (10), 917 - 921, 日本語


• 最新のテクノロジーが切り拓く骨代謝研究 生体イメージングと骨代謝

  菊田 順一, 石井 優

  (一社)日本骨代謝学会, 2021年10月, 日本骨代謝学会学術集会プログラム抄録集, 39回, 89 - 89, 日本語


• Rising Stars in Skeletal Biology SLPIはPTH誘導性骨形成に重要なメディエーターである

  森本 彬人, 菊田 順一, 石井 優

  (一社)日本骨代謝学会, 2021年10月, 日本骨代謝学会学術集会プログラム抄録集, 39回, 105 - 105, 日本語


• 【脳とからだ】腸,免疫系,脳の相互作用 交感神経系による骨髄の制御

  粟生 智香, 菊田 順一, 石井 優

  (公財)金原一郎記念医学医療振興財団, 2021年10月, 生体の科学, 72(5) (5), 393 - 396, 日本語


• 関節炎の炎症性骨破壊に関与する悪玉破骨前駆細胞の同定と機能

  揚村 朋弥, 長谷川 哲雄, 菊田 順一, 石井 優

  鳥居薬品(株), 2021年10月, 感染・炎症・免疫, 51(2) (2), 89 - 99, 日本語


• 【関節リウマチの診療update-最新の診断と治療-】関節リウマチの病態と治療薬の作用点

  鎗 伸弥, 菊田 順一, 石井 優

  (株)日本臨床社, 2021年09月, 日本臨床, 79(9) (9), 1272 - 1276, 日本語


• 骨のライブイメージング

  鎗 伸弥, 菊田 順一, 石井 優

  (株)ニュー・サイエンス社, 2021年06月, 細胞, 53(7) (7), 420 - 421, 日本語


• 【難治性免疫疾患-病態解明と新規治療戦略】免疫疾患の病態へのアプローチ マクロファージ研究によって解き明かされてきた難治性免疫疾患の実体

  宮本 佑, 菊田 順一, 石井 優

  医歯薬出版(株), 2021年05月, 医学のあゆみ, 277(9) (9), 682 - 688, 日本語


• 【膠原病・自己免疫疾患を「見える化」する】基礎から見える化する 骨関節破壊を見える化する

  鎗 伸弥, 菊田 順一, 石井 優

  (株)医学書院, 2020年11月, Medicina, 57(12) (12), 2145 - 2149, 日本語


• 臨床医が知っておくべき最新の基礎免疫学(第9回) 生体イメージング研究により明らかになったこと

  粟生 智香, 菊田 順一, 石井 優

  医歯薬出版(株), 2020年10月, 医学のあゆみ, 275(3) (3), 295 - 301, 日本語


• 生体イメージングによる細胞動態ネットワークの解明

  宮本 佑, 菊田 順一, 石井 優

  (公社)日本生化学会, 2020年10月, 生化学, 92(5) (5), 706 - 716, 日本語


• 骨免疫学研究最前線 骨免疫イメージングアップデート

  菊田 順一, 石井 優

  (一社)日本骨代謝学会, 2020年10月, 日本骨代謝学会学術集会プログラム抄録集, 38回, 97 - 97, 日本語


• 生体骨イメージングで解く破骨細胞の分化制御機構

  菊田 順一, 石井 優

  (一社)日本骨代謝学会, 2020年10月, 日本骨代謝学会学術集会プログラム抄録集, 38回, 103 - 103, 日本語


• SLPIは、骨芽細胞と破骨細胞間の相互作用を調節し、PTHによる骨形成を促進する

  森本 彬人, 石井 優, 菊田 順一

  (一社)日本骨代謝学会, 2020年10月, 日本骨代謝学会学術集会プログラム抄録集, 38回, 130 - 130, 日本語


• 局所の交感神経は急性炎症の初期において骨髄からの好中球の動員を促進する

  粟生 智香, 菊田 順一, 石井 優

  (一社)日本骨代謝学会, 2020年10月, 日本骨代謝学会学術集会プログラム抄録集, 38回, 142 - 142, 日本語


• 「骨の中の生きたままでのイメージング系」を駆使した新しい研究展開

  森本 彬人, 菊田 順一, 石井 優

  (有)科学評論社, 2020年09月, 臨床免疫・アレルギー科, 74(3) (3), 288 - 293, 日本語


• 骨破壊・免疫炎症の生体イメージング解析

  菊田 順一, 石井 優

  (一社)日本腎臓学会, 2020年09月, 日本腎臓学会誌, 62(6) (6), 503 - 503, 日本語


• 【細胞競合による生体制御とがん】細胞競合の力学・動態・数理 細胞競合と生体イメージング

  菊田 順一, 松井 崇浩, 石井 優

  医歯薬出版(株), 2020年08月, 医学のあゆみ, 274(5) (5), 531 - 536, 日本語


• 【線維化 慢性疾患のキープロセス 多彩な間質細胞が織りなす組織リモデリング"fibrosis"の理解】(第4章)線維化に対する次世代アプローチ 生体二光子励起イメージングによる線維化のin vivo解析

  宮本 佑, 菊田 順一, 石井 優

  (株)羊土社, 2020年08月, 実験医学, 38(12) (12), 2116 - 2121, 日本語


• 骨はどのようにして維持されるのか 相反する機能を持つ細胞による動的な制御機構

  鎗 伸弥, 菊田 順一, 石井 優

  (一社)日本生物物理学会, 2020年07月, 生物物理, 60(4) (4), 227 - 230, 日本語


• 骨はどのようにして維持されるのか:相反する機能を持つ細胞による動的な制御機構

  鎗伸弥, 菊田順一, 石井優

  2020年, 生物物理(Web), 60(4) (4)


• 骨芽細胞・破骨細胞は細胞間接触を介して骨リモデリングを調節する

  古家雅之, 古家雅之, 古家雅之, 菊田順一, 瀬尾茂人, 前田拓樹, 柏井将文, 柏井将文, 海渡貴司, 橋本国彦, 橋本国彦, 松田秀雄, 菊地和也, 石井優, 吉川秀樹, 吉川秀樹

  2020年, Journal of Spine Research (Web), 11(3) (3)


• 【新しい解析技術の展開】次世代の生体イメージング技術開発

  菊田 順一, 松井 崇浩, 石井 優

  (有)科学評論社, 2019年09月, 臨床免疫・アレルギー科, 72(3) (3), 293 - 298, 日本語


• 骨関節破壊の画像評価を極める 生体試料を見る

  菊田 順一, 石井 優

  (一社)日本骨代謝学会, 2019年09月, 日本骨代謝学会学術集会プログラム抄録集, 37回, 118 - 118, 日本語


• pH応答性蛍光プローブを用いた破骨細胞の骨吸収動態の解析

  菊田 順一, 石井 優

  (一社)日本骨代謝学会, 2019年09月, 日本骨代謝学会学術集会プログラム抄録集, 37回, 203 - 203, 日本語


• PTH製剤による骨形成促進メカニズムの網羅的解析

  森本 彬人, 菊田 順一, 石井 優

  (一社)日本骨代謝学会, 2019年09月, 日本骨代謝学会学術集会プログラム抄録集, 37回, 223 - 223, 日本語


• 骨関節破壊の画像評価を極める 生体試料を見る

  菊田 順一, 石井 優

  (一社)日本骨粗鬆症学会, 2019年09月, 日本骨粗鬆症学会雑誌, 5(Suppl.1) (Suppl.1), 206 - 206, 日本語


• 【"適応&修復"のサイエンスと臨床応用の最前線】様々な組織・臓器における適応・修復機構の動的イメージング

  菊田 順一, 石井 優

  (株)北隆館, 2019年07月, 別冊Bio Clinica: 慢性炎症と疾患, 8(1) (1), 10 - 14, 日本語


• 【運動器を構成する骨・軟骨・関節・筋肉・靱帯・腱のトピック】骨リモデリングに関する最新のトピック in vivoイメージングから得た新知見

  菊田 順一, 石井 優

  (株)メディカルレビュー社, 2019年05月, THE BONE, 33(1) (1), 51 - 55, 日本語


• 【4D細胞・組織ダイナミクス】イメージング技術と細胞・組織ダイナミクス

  宮本 佑, 菊田 順一, 石井 優

  (株)先端医学社, 2019年04月, 炎症と免疫, 27(3) (3), 192 - 198, 日本語


• 新しい解析技術と今後のリウマチ学の発展 生体イメージングによる疾患・創薬研究

  菊田 順一

  (一社)日本リウマチ学会, 2019年03月, 日本リウマチ学会総会・学術集会プログラム・抄録集, 63回, 228 - 228, 日本語


• 【ロコモの最新画像診断】骨の画像診断

  菊田 順一, 石井 優

  (株)先端医学社, 2018年11月, Loco Cure, 4(4) (4), 312 - 317, 日本語


• 生体骨イメージングによる生物学的製剤のin vivo作用機序の解明

  菊田 順一, 松浦 良信, 平野 亨, 熊ノ郷 淳, 石井 優

  (一社)日本臨床免疫学会, 2018年11月, 日本臨床免疫学会総会プログラム・抄録集, 46回, 104 - 104, 日本語


• 生体骨イメージングによる生物学的製剤のin vivo作用機序の解明

  菊田 順一, 松浦 良信, 平野 亨, 熊ノ郷 淳, 石井 優

  (一社)日本臨床免疫学会, 2018年11月, 日本臨床免疫学会総会プログラム・抄録集, 46回, 122 - 122, 日本語


• 骨関節破壊の生体イメージング

  菊田 順一, 石井 優

  (有)科学評論社, 2018年10月, リウマチ科, 60(4) (4), 442 - 446, 日本語


• 【アレルギーの分子的基盤研究の新しい展開】口腔粘膜特異的マクロファージサブセットが舌下免疫療法における免疫寛容の誘導に関与する

  青井 啓太, 菊田 順一, 石井 優

  (有)科学評論社, 2018年10月, 臨床免疫・アレルギー科, 70(4) (4), 357 - 360, 日本語


• 動態学-生きた動きを解析する新しい生命医科学 生体イメージングでみる骨代謝ダイナミクス

  菊田 順一

  (公社)日本生化学会, 2018年09月, 日本生化学会大会プログラム・講演要旨集, 91回, [2S04a - 04], 日本語


• 関節リウマチの関節破壊メカニズム 生体イメージングによる病態評価

  菊田 順一

  (公社)日本整形外科学会, 2018年08月, 日本整形外科学会雑誌, 92(8) (8), S1849 - S1849, 日本語


• 骨代謝・骨免疫(第17回) 生体骨組織イメージング

  菊田 順一, 石井 優

  (株)先端医学社, 2018年07月, 分子リウマチ治療, 11(3) (3), 144 - 147, 日本語


• Rising Stars in Skeletal Biology 二光子励起生体イメージングを用いた骨芽細胞・破骨細胞コミュニケーションの解明

  古家 雅之, 菊田 順一, 瀬尾 茂人, 前田 拓樹, 吉川 秀樹, 石井 優

  (一社)日本骨代謝学会, 2018年07月, 日本骨代謝学会学術集会プログラム抄録集, 36回, 117 - 117, 日本語


• 生体骨イメージング技術と計測解析の最前線 生体骨イメージングで見る破骨細胞・骨芽細胞間の4D相互作用

  古家 雅之, 菊田 順一, 瀬尾 茂人, 前田 拓樹, 吉川 秀樹, 石井 優

  日本骨形態計測学会, 2018年05月, 日本骨形態計測学会雑誌, 28(2) (2), S57 - S57, 日本語


• pH感受性赤色蛍光プローブを用いた破骨細胞プロトンポンプ動態のイメージング

  蓑島 維文, 大森 雄太, 菊田 順一, 石井 優, 菊地 和也

  日本分子イメージング学会, 2018年05月, JSMI Report, 11(2) (2), 70 - 70, 日本語


• 【イメージングの最前線】骨疾患のイメージング

  菊田 順一, 石井 優

  (有)科学評論社, 2018年04月, 臨床免疫・アレルギー科, 69(4) (4), 307 - 311, 日本語


• 【脂質メディエーターと病態】S1Pと骨病態

  菊田 順一, 石井 優

  (株)ニュー・サイエンス社, 2018年03月, 細胞, 50(3) (3), 136 - 139, 日本語


• イメージング技術が切り拓くリウマチ学の新展開 骨・関節の蛍光生体イメージングによる骨破壊の病態解明

  菊田 順一

  (一社)日本リウマチ学会, 2018年03月, 日本リウマチ学会総会・学術集会プログラム・抄録集, 62回, 251 - 251, 日本語


• 「アレルギーの臨床」に寄せる 骨・関節の生体イメージングによる骨破壊の病態解明

  菊田 順一, 石井 優

  (株)北隆館, 2018年02月, アレルギーの臨床, 38(2) (2), 196 - 198, 日本語


• 【運動器系の制御機構と破綻】可視化技術による運動器系組織のダイナミズム

  菊田 順一, 石井 優

  (株)医薬ジャーナル社, 2018年02月, Clinical Calcium, 28(3) (3), 367 - 371, 日本語


• 【リウマチ・膠原病の画像診断の進歩】生体イメージングで関節リウマチの病態を可視化する

  菊田 順一, 石井 優

  (株)先端医学社, 2018年01月, 分子リウマチ治療, 11(1) (1), 25 - 28, 日本語


• 【生体骨イメージング最前線～新しい創薬に向けて～】生体骨イメージングの基礎

  菊田 順一, 石井 優

  (株)医薬ジャーナル社, 2018年01月, Clinical Calcium, 28(2) (2), 175 - 179, 日本語


• 【生体骨イメージング最前線～新しい創薬に向けて～】生体骨イメージングの実際 破骨細胞

  菊田 順一, 石井 優

  (株)医薬ジャーナル社, 2018年01月, Clinical Calcium, 28(2) (2), 211 - 216, 日本語


• 【生体骨イメージング最前線～新しい創薬に向けて～】生体骨イメージングによる骨吸収抑制薬の作用解析

  菊田 順一, 石井 優

  (株)医薬ジャーナル社, 2018年01月, Clinical Calcium, 28(2) (2), 237 - 242, 日本語


• 【生体骨イメージング最前線～新しい創薬に向けて～】生体骨イメージングによる骨形成促進薬の作用解析

  森本 彬人, 菊田 順一, 古家 雅之, 石井 優

  (株)医薬ジャーナル社, 2018年01月, Clinical Calcium, 28(2) (2), 243 - 248, 日本語


• 生体イメージング技術による時空間情報を保持した1細胞遺伝子解析

  尾上 ひかる, 菊田 順一, 粟生 智香, 内田 穣, 石井 優

  生命科学系学会合同年次大会運営事務局, 2017年12月, 生命科学系学会合同年次大会, 2017年度, [3AT25 - 1442)], 日本語


• The effects of biologic agents on osteoclast lineage cells evaluated by intravital two-photon microscopy

  Yoshinobu Matsuura, Junichi Kikuta, Masaru Ishii

  2017年12月, CYTOKINE, 100, 111 - 112, 英語

  研究発表ペーパー・要旨（国際会議）


• 【ビタミンD Update】自己免疫疾患におけるビタミンD 基礎研究から臨床応用まで

  粟生 智香, 菊田 順一, 石井 優

  (株)医薬ジャーナル社, 2017年10月, Clinical Calcium, 27(11) (11), 1551 - 1559, 日本語


• 【免疫・炎症のin vivoイメージング】骨関節破壊のイメージング

  菊田 順一, 石井 優

  (一社)日本臨床免疫学会, 2017年10月, 日本臨床免疫学会会誌, 40(5) (5), 344 - 351, 日本語


• 骨代謝調節機構における新しい研究展開 生体イメージングで解く骨代謝調節機構

  菊田 順一

  (一社)歯科基礎医学会, 2017年09月, Journal of Oral Biosciences Supplement, 2017, 174 - 174, 日本語


• in vivoイメージングによる炎症解析 骨・関節炎症の生体イメージング

  菊田 順一

  (一社)日本臨床免疫学会, 2017年08月, 日本臨床免疫学会会誌, 40(4) (4), 268 - 268, 日本語


• 二光子励起生体イメージングを用いた破骨細胞・骨芽細胞コミュニケーションの解明

  古家 雅之, 菊田 順一, 瀬尾 茂人, 前田 拓樹, 柏井 史文, 海渡 貴司, 菊地 和也, 松田 秀雄, 吉川 秀樹, 石井 優

  (公社)日本整形外科学会, 2017年08月, 日本整形外科学会雑誌, 91(8) (8), S1711 - S1711, 日本語


• 【骨免疫学の進歩が変える骨関節疾患アプローチ】免疫細胞と骨代謝 免疫細胞による骨代謝制御 生体イメージング

  菊田 順一, 石井 優

  (株)メディカルレビュー社, 2017年07月, THE BONE, 31(2) (2), 151 - 155, 日本語


• Global Data Associationを用いたアメーバ状細胞の追跡手法 (情報論的学習理論と機械学習)

  間下 以大, 深野 淳, 白崎 舞, 菊田 順一, 石井 優, 竹村 治雄

  電子情報通信学会, 2017年06月23日, 電子情報通信学会技術研究報告 = IEICE technical report : 信学技報, 117(110) (110), 19 - 24, 日本語


• Global Data Associationを用いたアメーバ状細胞の追跡手法 (ニューロコンピューティング)

  間下 以大, 深野 淳, 白崎 舞, 菊田 順一, 石井 優, 竹村 治雄

  電子情報通信学会, 2017年06月23日, 電子情報通信学会技術研究報告 = IEICE technical report : 信学技報, 117(109) (109), 69 - 74, 日本語


• 「リウマチ・膠原病とアレルギー疾患」に寄せる 生体イメージング技術による骨・関節疾患の病態解明

  菊田 順一, 石井 優

  (株)北隆館, 2017年05月, アレルギーの臨床, 37(5) (5), 487 - 489, 日本語


• 電子顕微鏡・光学顕微鏡による最先端・骨イメージング 二光子励起顕微鏡で観る骨組織内の細胞動態ネットワーク

  菊田 順一

  日本骨形態計測学会, 2017年05月, 日本骨形態計測学会雑誌, 27(1) (1), S56 - S56, 日本語


• pH感受性赤色蛍光プローブを用いた生体内破骨細胞活性リアルタイムイメージング

  蓑島 維文, 大森 雄太, 前田 拓樹, 菊田 順一, 石井 優, 菊地 和也

  日本分子イメージング学会, 2017年05月, JSMI Report, 10(2) (2), 128 - 128, 日本語


• 細胞接着・接触を介した肝臓における糖代謝制御機構の解明

  宮地 康高, 土屋 恭一郎, 柴 久美子, 森 健太郎, 出牛 三千代, 大坂 瑞子, 吉田 雅幸, 菊田 順一, 石井 優, 小川 佳宏

  (一社)日本内分泌学会, 2017年04月, 日本内分泌学会雑誌, 93(1) (1), 380 - 380, 日本語


• 細胞間物理的相互作用に注目した肥満に伴う肝臓の糖代謝異常の分子機構

  宮地 康高, 土屋 恭一郎, 山口 忍, 柴 久美子, 森 健太郎, 出牛 三千代, 大坂 瑞子, 吉田 雅幸, 菊田 順一, 石井 優, 小川 佳宏

  (一社)日本糖尿病学会, 2017年04月, 糖尿病, 60(Suppl.1) (Suppl.1), S - 491, 日本語


• 緑内障手術のサイエンス 濾過手術とサイトカインを中心に

  井上 俊洋, 谷原 秀信, 布田 龍佑, 福島 美紀子, 伊藤 康裕, 川路 隆博, 岩尾 圭一郎, 高橋 枝里, 藤本 智和, 井上 みゆき, 岩尾 美奈子, 笠岡 奈々子, 原 竜平, 正林 耕平, 榮木 大輔, 大平 さおり, 後藤 章子, 小島 祥, 芳賀 彰, 黒田 詩子, 中島 圭一, 平川 沙織, 福島 亜矢子, 松村 理世, 徳永 瞳, 川畑 和幸, 北村 文香, 西澤 麻保, 二口 亜希子, 松本 桃佳, 吉村 長久, 亀田 隆範, 吉田 晃敏, 川井 基史, 川井 尚子, 木下 茂, 上田 真由美, 古賀 彩加, 稲谷 大, 瀧原 祐史, 本庄 恵, 石井 優, 菊田 順一, 古家 雅之, 松川 昭博, 猿渡 淳二, 田賀 哲也, 鹿川 哲史

  (公財)日本眼科学会, 2017年03月, 日本眼科学会雑誌, 121(3) (3), 314 - 335, 日本語


• 細胞接着と接触を介した肥満に伴う糖代謝異常の惹起機構

  宮地 康高, 土屋 恭一郎, 柴 久美子, 森 健太郎, 出牛 三千代, 大坂 瑞子, 吉田 雅幸, 菊田 順一, 石井 優, 小川 佳宏

  (一社)日本内分泌学会, 2017年01月, 日本内分泌学会雑誌, 92(3) (3), 889 - 889, 日本語


• 【骨粗鬆症治療における新しい接点】生体骨イメージングで捉える骨代謝

  菊田 順一, 古家 雅之, 石井 優

  (株)先端医学社, 2017年01月, 骨粗鬆症治療, 16(1) (1), 52 - 55, 日本語


• 【慢性炎症性疾患の新たな展開】慢性炎症の病的意義 骨代謝と慢性炎症

  菊田 順一, 松浦 良信, 石井 優

  (株)最新医学社, 2016年11月, 最新医学, 71(11月増刊) (11月増刊), 2257 - 2262, 日本語


• 【マクロファージのすべて】組織マクロファージ 破骨細胞

  菊田 順一, 石井 優

  医歯薬出版(株), 2016年10月29日, 医学のあゆみ, 259(5) (5), 446 - 452, 日本語


• 肝臓における細胞接触・接着を介した新たな糖代謝制御機構の解明

  宮地 康高, 土屋 恭一郎, 柴 久美子, 森 健太郎, 出牛 三千代, 大坂 瑞子, 吉田 雅幸, 菊田 順一, 石井 優, 小川 佳宏

  (一社)日本肥満学会, 2016年09月, 肥満研究, 22(Suppl.) (Suppl.), 193 - 193, 日本語


• 病理アトラス 膠原病・アレルギー疾患の可視化

  菊田 順一, 石井 優

  (株)北隆館, 2016年08月, 別冊Bio Clinica: 慢性炎症と疾患, 5(3) (3), 1 - 4, 日本語


• 免疫・炎症のin vivoイメージング 骨関節破壊のイメージング

  菊田 順一

  (一社)日本臨床免疫学会, 2016年08月, 日本臨床免疫学会会誌, 39(4) (4), 351 - 351, 日本語


• 運動器イメージングの進歩 蛍光生体イメージングで解く破骨細胞・骨芽細胞のクロストーク

  菊田 順一

  (公社)日本整形外科学会, 2016年08月, 日本整形外科学会雑誌, 90(8) (8), S1435 - S1435, 日本語


• 【骨リモデリングの制御機構】骨芽細胞と破骨細胞の生体イメージング観察

  菊田 順一, 古家 雅之, 石井 優

  (株)メディカルレビュー社, 2016年07月, THE BONE, 30(2) (2), 141 - 144, 日本語


• 先端技術で読み解く正常造血と造血器腫瘍 蛍光生体イメージングで解く免疫・血液細胞の動的ネットワーク

  菊田 順一

  (一社)日本サイトメトリー学会, 2016年07月, Cytometry Research, 26(Suppl.) (Suppl.), 36 - 36, 日本語


• 汗腺の構造/機能解析の最前線 Claudin-3が汗腺からの汗の漏出を止める

  山賀 康右, 室田 浩之, 宮田 浩史, 近江 雅人, 菊田 順一, 石井 優, 田村 淳, 月田 早智子, 片山 一朗

  日本発汗学会, 2016年07月, 日本発汗学会総会プログラム・抄録集, 24回, 31 - 31, 日本語


• 骨免疫学の最前線 骨免疫イメージング

  菊田 順一

  (一社)日本骨代謝学会, 2016年07月, 日本骨代謝学会学術集会プログラム抄録集, 34回, 115 - 115, 日本語


• 骨小腔・骨細管のin vivoイメージング

  佐野 博繁, 菊田 順一, 古家 雅之, 近藤 直樹, 藤澤 純一, 木島 靖文, 遠藤 直人, 石井 優

  日本骨形態計測学会, 2016年06月, 日本骨形態計測学会雑誌, 26(1) (1), S148 - S148, 日本語


• 病態メカニズムに迫るイメージング技術 DDSへの期待 生体二光子励起イメージングによる骨髄血管透過性の制御機構解明とDDS研究への応用

  菊田 順一

  日本DDS学会, 2016年06月, 日本DDS学会学術集会プログラム予稿集, 32回, 92 - 92, 日本語


• Physical Cell-Cell Interaction Regulates Hepatic Glucose Metabolism in Mice

  Kyoichiro Tsuchiya, Yasutaka Miyachi, Chikara Komiya, Kumiko Shiba, Noriko Shimazu, Shinobu Yamaguchi, Mizuko Osaka, Masayuki Yoshida, Koji Inoue, Kenjiro Wake, Yuta Sato, Junichi Kikuta, Masaru Ishii, Yoshihiro Ogawa

  2016年06月, DIABETES, 65, A462 - A462, 英語

  研究発表ペーパー・要旨（国際会議）


• 最新用語解説 基礎(第52回) 生体イメージング

  菊田 順一, 石井 優

  (株)先端医学社, 2016年06月, 骨粗鬆症治療, 15(1) (1), 59 - 63, 日本語


• ウェーブレット特徴量を用いた生体骨組織の骨髄腔の領域分割手法

  繁田浩功, 間下以大, 菊田順一, 瀬尾茂人, 竹村治雄, 松田秀雄, 石井優

  2016年06月, 日本語


• 慢性炎症時のマクロファージ動態のイメージング解析

  菊田 順一, 石井 優

  (株)最新医学社, 2016年05月, 最新医学, 71(5) (5), 1026 - 1030, 日本語


• 【代謝調節における免疫細胞の役割】最新の解析技術 免疫細胞の生体イメージング

  菊田 順一, 石井 優

  医歯薬出版(株), 2016年05月07日, 医学のあゆみ, 257(6) (6), 727 - 731, 日本語


• pH感受性蛍光プローブを用いた生体内破骨細胞活性リアルタイムイメージング

  大森 雄太, 前田 拓樹, 小和田 俊行, 菊田 順一, 古家 雅之, 石井 優, 菊地 和也

  日本分子イメージング学会, 2016年04月, JSMI Report, 9(2) (2), 134 - 134, 日本語


• 【慢性炎症と自己免疫疾患】生体イメージングで捉える免疫炎症・骨破壊の動的な実体

  菊田 順一, 石井 優

  (一社)日本臨床免疫学会, 2016年04月, 日本臨床免疫学会会誌, 39(2) (2), 124 - 129, 日本語


• 骨免疫研究の最前線 破骨細胞動態イメージング

  菊田 順一

  (一社)日本リウマチ学会, 2016年03月, 日本リウマチ学会総会・学術集会プログラム・抄録集, 60回, 176 - 176, 日本語


• 【再生医学と血管】血管生物学における生体イメージング技術

  菊田 順一, 石井 優

  (株)メディカルレビュー社, 2016年03月, 血管医学, 17(1) (1), 41 - 45, 日本語


• 生体イメージング技術を駆使したトロンボモジュリンの新たな作用機序解明

  西澤 志乃, 菊田 順一, 瀬尾 茂人, 松田 秀雄, 石井 優

  (公社)日本生化学会, 2015年12月, 日本生化学会大会・日本分子生物学会年会合同大会講演要旨集, 88回・38回, [3T23p - 05(3P0080)], 日本語


• 【骨関節疾患の画像評価】In vivoイメージング 骨関節疾患評価の新たな取り組み

  菊田 順一, 石井 優

  (株)先端医学社, 2015年12月, Rheumatology Clinical Research, 4(3) (3), 180 - 185, 日本語


• 最新基礎科学 知っておきたい ビタミンDの骨吸収抑制機序

  菊田 順一, 石井 優

  (株)医学書院, 2015年11月, 臨床整形外科, 50(11) (11), 1114 - 1117, 日本語


• IL-6から見える景色(第9回) IL-6と骨破壊 反応編(4)

  松浦 良信, 菊田 順一, 石井 優

  (株)先端医学社, 2015年10月, Keynote R・A, 3(4) (4), 190 - 191, 日本語


• 多光子励起顕微鏡を用いた骨組織細胞動態イメージング

  菊田 順一, 石井 優

  (有)科学評論社, 2015年08月, 臨床免疫・アレルギー科, 64(2) (2), 194 - 199, 日本語


• 骨の4Dイメージング 骨のライブイメージング最前線

  菊田 順一, 水野 紘樹, 古家 雅之, 石井 優

  (株)メディカルレビュー社, 2015年08月, O.li.v.e., 5(3) (3), 208 - 211, 日本語


• 最新用語解説 基礎(第50回) S1P

  菊田 順一, 石井 優

  (株)先端医学社, 2015年08月, 骨粗鬆症治療, 14(2) (2), 151 - 154, 日本語


• in vivoイメージングを用いた骨細胞性骨溶解機序解明への試み

  佐野 博繁, 菊田 順一, 古家 雅之, 近藤 直樹, 遠藤 直人, 石井 優

  (一社)日本骨代謝学会, 2015年07月, 日本骨代謝学会学術集会プログラム抄録集, 33回, 166 - 166, 日本語


• Imaging and hematopoiesis/oncogenesis 生体骨組織を用いた多光子励起イメージングによるin vivo造血細胞の動態解析

  水野 紘樹, 菊田 順一, 古家 雅之, 石井 優

  (一社)日本血液学会-東京事務局, 2015年06月, 臨床血液, 56(6) (6), 594 - 600, 日本語


• 【骨の生体イメージング 最前線】破骨細胞の生体イメージング

  菊田 順一, 石井 優

  (株)医薬ジャーナル社, 2015年05月, Clinical Calcium, 25(6) (6), 815,789 - 790, 日本語


• 骨の4Dイメージング

  菊田 順一, 石井 優

  (株)メディカルレビュー社, 2015年05月, O.li.v.e., 5(2) (2), 132 - 135, 日本語


• 【骨転移病変に対する最新の基礎的知見と臨床マネージメント】固形癌転移と骨微小環境をめぐる最新の基礎研究 破骨細胞イメージングによる骨破壊メカニズムの解明

  菊田 順一, 石井 優

  (株)メディカルレビュー社, 2015年03月, Surgery Frontier, 22(1) (1), 28,3 - 4, 日本語


• 【ビタミンDをめぐる最近の話題】ビタミンDの免疫細胞への作用と感染防御

  菊田 順一, 石井 優

  (株)医薬ジャーナル社, 2015年02月, Clinical Calcium, 25(3) (3), 359 - 365, 日本語


• 病理アトラス 生体イメージングによる炎症病態の機能的解析

  水野 紘樹, 菊田 順一, 石井 優

  (株)北隆館, 2015年02月, 別冊Bio Clinica: 慢性炎症と疾患, 4(1) (1), np1 - np4, 日本語


• PAMPs受容体研究の最前線 スフィンゴシン1リン酸受容体とB細胞受容体との会合にMD-1は影響を及ぼす(PAMPs receptor update MD-1 influences a binding between S1P1 and BCR)

  高村 祥子, 山川 奈津子, 谷村 奈津子, 柴田 琢磨, 鈴木 一博, 菊田 順一, 石井 優, 三宅 健介

  日本細菌学会, 2015年02月, 日本細菌学雑誌, 70(1) (1), 128 - 128, 英語


• 【骨代謝研究の最前線】イメージングを用いた骨代謝研究

  菊田 順一, 石井 優

  (株)北隆館, 2015年01月, BIO Clinica, 30(1) (1), 23 - 27, 日本語


• 内分泌 基礎分野での進歩 骨代謝のin vivoイメージング

  菊田 順一, 石井 優

  (株)中外医学社, 2015年01月, Annual Review糖尿病・代謝・内分泌, 2015, 152 - 159, 日本語


• 骨の生体二光子励起イメージング 骨代謝・免疫研究の新展開

  菊田 順一

  松本歯科大学学会, 2014年12月, 松本歯学, 40(2) (2), 170 - 171, 日本語


• 【炎症-全体像を知り慢性疾患を制御する 炎症シグナル、免疫細胞のダイナミズムからがん・糖尿病などの発症機序と治療標的まで】(第1章)炎症応答の時・空間的ダイナミクスと関与する細胞たち 炎症のvisualization

  菊田 順一, 石井 優

  (株)羊土社, 2014年11月, 実験医学, 32(17) (17), 2716 - 2720, 日本語


• 【がん・免疫・代謝・発生におけるM1/M2分類を超えたマクロファージの機能】ライブイメージングから見る骨吸収・慢性炎症時のマクロファージ動態

  菊田 順一, 石井 優

  (株)学研メディカル秀潤社, 2014年11月, 細胞工学, 33(12) (12), 1267 - 1271, 日本語


• 骨と骨髄による全身制御 生体二光子励起イメージングによる骨代謝ダイナミクスの動的解析

  菊田 順一

  (公社)日本生化学会, 2014年10月, 日本生化学会大会プログラム・講演要旨集, 87回, [2S12p - 4], 日本語


• リウマチ治療薬の効果予測及び効果判定を目的としたIL-6受容体抗体の125I標識及び体内動態の観察

  金井 泰和, 菊田 順一, 渡部 直史, 仲 定宏, 礒橋 佳也子, 加藤 弘樹, 巽 光朗, 阿部 浩司, 下瀬川 恵久, 石井 優, 畑澤 順

  (一社)日本核医学会, 2014年09月, 核医学, 51(3) (3), 336 - 336, 英語


• 目で見るBone Biology(第38回) S1P/S1P受容体と骨粗鬆症

  菊田 順一, 石井 優

  (株)先端医学社, 2014年08月, 骨粗鬆症治療, 13(2) (2), 93 - 96, 日本語


• 【高齢者脆弱性骨折の予防と治療】病態の最新知見 画像イメージングから

  菊田 順一, 古家 雅之, 石井 優

  (株)南江堂, 2014年07月, 整形外科, 65(8) (8), 712 - 718, 日本語


• 新たなイメージングの展開 生体多光子励起イメージングによる生きた細胞動態の解析

  菊田 順一

  (一社)日本医用画像工学会, 2014年07月, MEDICAL IMAGING TECHNOLOGY, 32(Suppl.) (Suppl.), 1 - 1, 日本語


• 骨代謝研究領域におけるバイオイメージング技術の発展からその応用まで 2光子励起顕微鏡を駆使した骨髄イメージング

  菊田 順一, 石井 優

  (一社)日本骨代謝学会, 2014年07月, 日本骨代謝学会学術集会プログラム抄録集, 32回, 160 - 160, 日本語


• 生体二光子励起イメージングによるビスホスホネート製剤の短期的効果の検討

  菊田 順一, 白崎 舞, 石井 優

  (一社)日本骨代謝学会, 2014年07月, 日本骨代謝学会学術集会プログラム抄録集, 32回, 205 - 205, 日本語


• 【骨バイオイメージング】動的イメージング 骨髄内細胞動態イメージング(2光子励起顕微鏡を用いて)

  菊田 順一, 石井 優

  (株)メディカルレビュー社, 2014年06月, THE BONE, 28(2) (2), 185 - 190, 日本語


• 【骨・血管連関～最近の進歩～】免疫系と骨・血管連関

  菊田 順一, 石井 優

  (株)医薬ジャーナル社, 2014年06月, Clinical Calcium, 24(7) (7), 1031 - 1036, 日本語


• 新しい医療技術 4Dイメージング

  菊田 順一, 石井 優

  金原出版(株), 2014年06月, 整形・災害外科, 57(7) (7), 921 - 926, 日本語


• 【骨代謝 つくり、壊し、変える-そのメカニズムと最新治療 分子機構から骨粗鬆症・リウマチなど骨疾患への応用まで】(第I部) 骨代謝制御機構を知る (第3章)骨をみる イメージングの現状と展望 骨組織におけるin vivoイメージング

  菊田 順一, 古家 雅之, 石井 優

  (株)羊土社, 2014年05月, 実験医学, 32(7) (7), 1054 - 1060, 日本語


• 【最新関節リウマチ学-寛解・治癒を目指した研究と最新治療-】関節リウマチの基礎研究 骨組織の生体多光子励起イメージング

  菊田 順一, 石井 優

  (株)日本臨床社, 2014年04月, 日本臨床, 72(増刊3 最新関節リウマチ学) (増刊3 最新関節リウマチ学), 178 - 182, 日本語


• 【骨と幹細胞】骨髄環境のライブイメージング

  水野 紘樹, 菊田 順一, 石井 優

  (株)医薬ジャーナル社, 2014年03月, Clinical Calcium, 24(4) (4), 541 - 546, 日本語


• 骨粗鬆症と骨代謝 活性型ビタミンDはS1Pによる破骨前駆細胞の遊走制御機構を調整することにより骨吸収を抑制する

  菊田 順一, 石井 優

  (一社)日本リウマチ学会, 2014年03月, 日本リウマチ学会総会・学術集会プログラム・抄録集, 58回, 387 - 387, 日本語


• グラフカットを用いた骨髄腔画像の領域分割

  宇佐見 潤, 繁田 浩功, 間下 以大, 黒田 嘉宏, 菊田 順一, 瀬尾 茂人, 石井 優, 松田 秀雄, 竹村 治雄

  生体イメージング技術の向上により生体内の細胞の動態を動画像として観察が可能となり，疾病のメカニズム解明や創薬等への応用が期待されている．これらの応用のためには，細胞画像から特定領域を抽出したり，細胞の特定の動きを検出する必要がある．また，膨大な数の画像に対して一定の基準で領域分割や細胞の検出を行うためには計算機での処理が必要である．本論文では，二光子励起顕微鏡を用いた骨髄腔画像の骨髄腔の領域分割手法を提案する．本研究で対象とする生体細胞画像は生体の一部を時系列画像として撮影したものであり，骨髄腔領域と血流領域を分割するには，輝度値の高低や血流の有無等の情報を総合して判断される．提案手法ではこれらの情報をエネルギー関数に取り入れた上でグラフカットによる最適化を行い，領域分割を実現する．提案手法と従来のグラフカットとの比較評価を行った結果，hard-constraint というユーザの入力に依存した制約を用いていない提案手法が hard-constraint を用いている従来手法と同等以上の性能が得られることを確認した．Emerging bio-imaging technologies are expected to contribute to the discovery of new drugs and the mechanisms by which diseases survive. In applications involving cell and bacterial imaging, extracting a particular region or detecting cell motion is essential. Moreover, automatic extraction and detection in image processing are also required because it is unrealistic to manually process a large number of images accurately, uniformly, and in a short period of time. To help automate this process, we introduce a bone-marrow space segmentation method for two-photon excitation microscopy images. Cellular dynamics specialists typically separate regions of bone-marrow and other spaces using several criteria such as blood flow characteristics and intensity. Taking these consideration, we designed data-term in graph-cuts method to process sequential images of the inside of a living mouse. Results of evaluations and comparison with normal graph-cuts show that our proposed method, which doesn't use hard constraints, achieves better performance than normal hard-constraint based graph-cuts methods.

  一般社団法人情報処理学会, 2014年02月24日, 研究報告数理モデル化と問題解決（MPS）, 2014(17) (17), 1 - 6, 日本語


• 【細胞の3次元組織化-その最先端技術と材料技術 再生医療とその支援分野(細胞研究、創薬研究)への応用と発展のために】(第4章)細胞3次元組織化培養のための周辺技術 ライブイメージング

  水野 紘樹, 菊田 順一, 石井 優

  (株)メディカルドゥ, 2014年02月, 遺伝子医学MOOK, 別冊(細胞の3次元組織化-その最先端技術と材料技術) (細胞の3次元組織化-その最先端技術と材料技術), 354 - 358, 日本語


• Dynamic analysis of short-term effects of bisphosphonates by using intravital two-photon microscopy

  Junichi Kikuta, Mai Shirazaki, Masaru Ishii

  2014年02月, JOURNAL OF BONE AND MINERAL RESEARCH, 29, S193 - S193, 英語

  研究発表ペーパー・要旨（国際会議）


• 免疫細胞の動態制御とストローマ細胞 生体多光子励起イメージングによる骨髄・免疫細胞の動態解析

  菊田 順一

  日本リンパ学会, 2013年12月, リンパ学, 36(2) (2), 123 - 126, 日本語


• 【ストレスによる骨制御】骨吸収のダイナミクス ライブイメージング

  菊田 順一, 石井 優

  (株)医薬ジャーナル社, 2013年10月, Clinical Calcium, 23(11) (11), 1627 - 1633, 日本語


• 【骨粗鬆症領域の進歩】破骨細胞研究の最前線

  菊田 順一, 石井 優

  (株)北隆館, 2013年09月, BIO Clinica, 28(10) (10), 911 - 916, 日本語


• 消化器研究における最新技術(第5回) 4Dイメージングによる消化器疾患への応用

  前田 栄, 菊田 順一, 石井 優

  (株)先端医学社, 2013年08月, G.I.Research, 21(4) (4), 368 - 374, 日本語


• 生体多光子励起イメージングによる「骨形態・動態計測」研究の最前線

  石井 優, 菊田 順一

  日本骨形態計測学会, 2013年06月, 日本骨形態計測学会雑誌, 23(1) (1), S71 - S71, 日本語


• 関節リウマチと骨軟骨代謝 バイオイメージングで解析する関節リウマチの骨破壊病態

  石井 優, 菊田 順一

  (一社)日本骨代謝学会, 2013年05月, 日本骨代謝学会学術集会プログラム抄録集, 31回, 66 - 66, 日本語


• 生体多光子励起顕微鏡による生体骨組織内の血管透過性の評価とその制御機構の解明

  金子 雄, 菊田 順一, 石井 優

  (一社)日本骨代謝学会, 2013年05月, 日本骨代謝学会学術集会プログラム抄録集, 31回, 104 - 104, 日本語


• Histamine attenuates acetylcholine-mediated sweating by inactivating GSK3 beta via H1-receptor mediated pathway

  S. Matsui, H. Murota, Y. Lingli, K. Omiya, M. Ohmi, J. Kikuta, M. Ishii, I. Katayama

  2013年05月, JOURNAL OF INVESTIGATIVE DERMATOLOGY, 133, S105 - S105, 英語

  研究発表ペーパー・要旨（国際会議）


• 【骨代謝-「見えざる手」が制御する骨破壊と骨形成 リウマチ・骨粗鬆症の発症機構と分子標的治療】骨組織のin vivoイメージング

  菊田 順一, 石井 優

  (株)羊土社, 2013年04月, 実験医学, 31(6) (6), 862 - 869, 日本語


• グラフカットを用いた生体骨組織における血管透過性の評価手法

  繁田 浩功, 間下 以大, 金子 雄, 菊田 順一, 瀬尾 茂人, 竹村 治雄, 松田 秀雄, 石井 優

  生体イメージング技術の向上により生体内の動態を画像として観察することが可能となり,疾病のメカニズム解明や創薬等への応用が期待されている.一方で統計的信頼性を得るために膨大な画像を解析する必要性が生じている.本研究では,生体骨組織内の血管透過性の制御機構を解明するため,二光子励起顕微鏡を用いて得られた生体画像に対して,血液が骨髄腔に染み出す血管透過性を評価する手法を提案する.提案手法では,対象となる時系列画像に対してグラフカットを用いて骨髄腔領域をセグメンテーションし,その染み出し量を定量的に評価する.評価実験の結果,専門家の手による抽出結果を用いた結果とほぼ同様の結果が得られた.このことから,本手法は染み出しの程度について統計的かつ定量的な評価の一つとして利用できると思われる.

  一般社団法人情報処理学会, 2013年03月14日, 情報処理学会研究報告. BIO, バイオ情報学, 2013(3) (3), 1 - 6, 日本語


• VF-022-2 生体イメージングによる細胞周期と浸潤関連分子の同定と臨床応用(VF ビデオフォーラム,第113回日本外科学会定期学術集会)

  賀川 義規, 前田 栄, 深見 美和, 菊田 順一, 内藤 敦, 西川 恵三, 太田 勝也, 藤森 さゆ美, Simmons Szandor, 石井 泰子, 植村 守, 原口 直紹, 西村 潤一, 畑 泰司, 竹政 伊知朗, 水島 恒和, 山本 浩文, 石井 秀始, 土岐 祐一郎, 石井 優, 森 正樹

  一般社団法人日本外科学会, 2013年03月05日, 日本外科学会雑誌, 114(2) (2), 440 - 440, 日本語


• 生体イメージングによる細胞周期と浸潤関連分子の同定と臨床応用

  賀川 義規, 前田 栄, 深見 美和, 菊田 順一, 内藤 敦, 西川 恵三, 太田 勝也, 藤森 さゆ美, Simmons Szandor, 石井 泰子, 植村 守, 原口 直紹, 西村 潤一, 畑 泰司, 竹政 伊知朗, 水島 恒和, 山本 浩文, 石井 秀始, 土岐 祐一郎, 石井 優, 森 正樹

  (一社)日本外科学会, 2013年03月05日, 日本外科学会雑誌, 114(臨増2) (臨増2), 440 - 440, 日本語


• 【骨評価法の進歩】蛍光イメージングによる骨評価

  菊田 順一, 石井 優

  (株)医薬ジャーナル社, 2013年02月, Clinical Calcium, 23(3) (3), 355 - 360, 日本語


• S1P-mediated Osteoclast Precursor Monocyte Migration Is a Critical Point of Control in Antibone-resorptive Action of Active Vitamin D

  Junichi Kikuta, Junichi Kikuta, Fumie Okiji, Mai Shirazaki, Sadaoki Sakai, Hitoshi Saito, Masaru Ishii

  2013年02月, JOURNAL OF BONE AND MINERAL RESEARCH, 28, 英語

  研究発表ペーパー・要旨（国際会議）


• Sphingosine-1-phosphate signaling controlling osteoclasts and bone homeostasis

  Masaru Ishii, Junichi Kikuta

  2013年01月, BIOCHIMICA ET BIOPHYSICA ACTA-MOLECULAR AND CELL BIOLOGY OF LIPIDS, 1831(1) (1), 223 - 227, 英語

  書評論文,書評,文献紹介等


• Orthovisual 骨のライブイメージング

  菊田 順一, 石井 優

  (株)メディカルレビュー社, 2012年12月, Arthritis-運動器疾患と炎症-, 10(3) (3), 170 - 176, 日本語


• 生体イメージングクリニック

  菊田 順一, 佐藤 康彦, 石井 優

  (NPO)日本免疫学会, 2012年11月, 日本免疫学会総会・学術集会記録, 41, 223 - 223, 日本語


• Dynamic in Vivo Imaging of Th17-Mediated Osteoclastic Bone Resorption in Live Bones by Using Intravital Multiphoton Microscopy

  Junichi Kikuta, Masaru Ishii

  2012年10月, ARTHRITIS AND RHEUMATISM, 64(10) (10), S311 - S312, 英語

  研究発表ペーパー・要旨（国際会議）


• 【骨免疫学-研究最前線】骨免疫とin vivoイメージング 螢光イメージングを用いた免疫細胞と破骨細胞の挙動

  菊田 順一, 石井 優

  医歯薬出版(株), 2012年09月01日, 医学のあゆみ, 242(9) (9), 727 - 733, 日本語


• 【血管系のin vivoイメージング】生体骨組織内での破骨細胞動態イメージング

  小谷 真奈斗, 菊田 順一, 石井 優

  (株)メディカルレビュー社, 2012年06月, 血管医学, 13(2) (2), 137 - 143, 日本語


• Evolving FACS Technology FACSを応用した生体イメージング関連分子の解析

  賀川 義規, 西川 恵三, 前田 栄, 菊田 順一, 太田 勝也, 西村 潤一, 竹政 伊知朗, 水島 恒和, 池田 正孝, 山本 浩文, 関本 貢嗣, 石井 秀始, 土岐 祐一郎, 森 正樹, 石井 優

  (一社)日本サイトメトリー学会, 2012年06月, Cytometry Research, 22(Suppl.) (Suppl.), 46 - 46, 日本語


• 癌の細胞周期蛍光イメージングによる抗腫瘍薬の効果ポイントの可視化

  賀川 義規, 前田 栄, 太田 勝也, 菊田 順一, 西川 恵三, 西村 潤一, 竹政 伊知朗, 水島 恒和, 池田 正孝, 山本 浩文, 関本 貢嗣, 石井 秀始, 土岐 祐一郎, 森 正樹, 石井 優

  (一社)日本サイトメトリー学会, 2012年06月, Cytometry Research, 22(Suppl.) (Suppl.), 57 - 57, 日本語


• DYNAMIC IN VIVO IMAGING OF TH17-MEDIATED CONTROL OF OSTEOCLAST FUNCTION IN LIVE BONES BY USING INTRAVITAL MULTIPHOTON MICROSCOPY

  J. Kikuta, M. Ishii

  2012年06月, ANNALS OF THE RHEUMATIC DISEASES, 71, 66 - 66, 英語

  研究発表ペーパー・要旨（国際会議）


• 【破骨細胞の機能とその異常】破骨細胞機能のイメージング

  菊田 順一, 石井 優

  (株)日本メディカルセンター, 2012年04月, 腎と骨代謝, 25(2) (2), 107 - 113, 日本語


• 関節リウマチの病因・病態(2) 生体多光子励起イメージングによる成熟破骨細胞の動態解析

  菊田 順一, 石井 優

  (一社)日本リウマチ学会, 2012年03月, 日本リウマチ学会総会・学術集会・国際リウマチシンポジウムプログラム・抄録集, 56回・21回, 341 - 341, 日本語


• 破骨細胞のイメージング

  菊田 順一, 石井 優

  (有)科学評論社, 2012年02月, リウマチ科, 47(2) (2), 195 - 199, 日本語


• Migratory control of bone-resorbing osteoclasts visualized by intravital multiphoton microcopy

  Masaru Ishii, Junichi Kikuta

  2012年, JOURNAL OF PHARMACOLOGICAL SCIENCES, 118, 23P - 23P, 英語

  研究発表ペーパー・要旨（国際会議）


• Dynamic in vivo imaging of bone-resorptive functions of mature osteoclasts in live bones by using intravital multiphoton microscopy

  Junichi Kikuta, Yoh Wada, Masaru Ishii

  2012年, JOURNAL OF PHARMACOLOGICAL SCIENCES, 118, 70P - 70P, 英語

  研究発表ペーパー・要旨（国際会議）


• 【分子イメージングの最先端】骨組織のライブイメージングによる破骨前駆細胞の動態解析

  小谷 真奈斗, 菊田 順一, 石井 優

  先端医療技術研究所, 2011年12月, PET Journal, (16) (16), 17 - 19, 日本語


• 【骨・カルシウムUPDATE】骨代謝研究UPDATE 蛍光イメージング技術で観察する破骨前駆細胞のトラフィッキング

  小谷 真奈斗, 菊田 順一, 石井 優

  (株)医薬ジャーナル社, 2011年11月, Clinical Calcium, 21(12) (12), 1835 - 1841, 日本語


• 【分子イメージングの最先端】内科疾患の分子イメージング 骨・免疫系の細胞動態イメージング

  島津 裕, 菊田 順一, 久保 厚子, 石井 優

  (株)最新医学社, 2011年10月, 最新医学, 66(10) (10), 2349 - 2353, 日本語


• New advances in allergy and immune regulation by imaging analysis 骨組織の生体多光子励起イメージングによる破骨細胞動態の解析

  石井 優, 菊田 順一

  (一社)日本アレルギー学会, 2011年10月10日, アレルギー, 60(9-10) (9-10), 1258 - 1258, 日本語


• 【RANKLシグナルと骨病変】破骨細胞のイメージングとRANKLシグナル

  菊田 順一, 石井 優

  (株)医薬ジャーナル社, 2011年07月, Clinical Calcium, 21(8) (8), 1181 - 1185, 日本語


• 生体二光子励起顕微鏡による成熟破骨細胞の「機能」のイメージング

  菊田 順一, 石井 優

  (一社)日本骨代謝学会, 2011年07月, 日本骨代謝学会学術集会プログラム抄録集, 29回, 197 - 197, 日本語


• 二光子励起顕微鏡を用いたがん細胞の生体イメージング

  賀川 義規, 石井 秀始, 石井 泰子, 久保 厚子, 鈴木 陽三, 宮崎 進, 川村 俊輔, 小谷 真奈斗, 藤森 さゆ美, 島津 裕, 菊田 順一, 竹政 伊知朗, 水島 恒和, 池田 正孝, 山本 浩文, 坂上 朝子[沢野], 宮脇 敦史, 石井 優, 関本 貢嗣, 土岐 祐一郎, 森 正樹

  (一社)日本外科学会, 2011年05月, 日本外科学会雑誌, 112(臨増1-2) (臨増1-2), 379 - 379, 日本語


• 【癌と生体イメージング】二光子励起顕微鏡による骨髄・骨転移性癌の生体イメージング

  小谷 真奈斗, 菊田 順一, 大畑 絵美, 石井 優

  (株)メディカルレビュー社, 2011年03月, Surgery Frontier, 18(1) (1), 44 - 49, 日本語


• 【がんと骨の遭遇】骨組織のin vivoイメージングによる破骨細胞動態の解析

  菊田 順一, 川村 俊輔, 石井 優

  (株)医薬ジャーナル社, 2011年02月, Clinical Calcium, 21(3) (3), 372 - 378, 日本語


• 破骨細胞遊走の制御機構

  菊田 順一, 小谷 真奈斗, 石井 優

  (有)科学評論社, 2011年02月, リウマチ科, 45(2) (2), 209 - 214, 日本語


• 【骨を壊して骨を作る 細胞レベルの制御から3Dデザインへ】蛍光イメージングによる骨解析

  菊田 順一, 川村 俊輔, 石井 優

  (株)学研メディカル秀潤社, 2011年02月, 細胞工学, 30(3) (3), 277 - 282, 日本語


• Chemokine-mediated migration control of bone cells visualized by intravital multiphoton bone imaging

  Masaru Ishii, Junichi Kikuta, Atsuko Kubo, Yutaka Shimazu

  2011年, JOURNAL OF PHARMACOLOGICAL SCIENCES, 115, 57P - 57P, 英語

  研究発表ペーパー・要旨（国際会議）


• Dynamic in vivo imaging of differentiation and function of osteoclasts in live bones by using intravital two-photon microscopy

  Junichi Kikuta, Issei Nishiyama, Atsuko Kubo, Yutaka Shimazu, Yoh Wada, Masaru Ishii

  2011年, JOURNAL OF PHARMACOLOGICAL SCIENCES, 115, 93P - 93P, 英語

  研究発表ペーパー・要旨（国際会議）


• スフィンゴシン1リン酸の破骨細胞分化に及ぼす影響

  天野 均, 唐川 亜希子, 菊田 順一, 石井 優, 山田 庄司

  (一社)日本骨代謝学会, 2010年07月, 日本骨代謝学会学術集会プログラム抄録集, 28回, 167 - 167, 日本語


• 【歯と骨の炎症制御の新規標的】骨組織の生体イメージングによる骨吸収の新しい調節機構

  菊田 順一, 石井 優

  (株)先端医学社, 2010年06月, 炎症と免疫, 18(4) (4), 349 - 354, 日本語


• 破骨細胞の遊走・位置決めの機構の解明とこれをターゲットとした新しい骨粗鬆症治療薬の開発

  石井 優, 岩井 香織, 菊田 順一

  ライフサイエンス出版(株), 2009年10月, Osteoporosis Japan, 17(4) (4), 689 - 691, 日本語


• 【生物学的DMARDs 抗リウマチ薬のイノベーション】RA治療におけるエビデンス 抗IL-6受容体抗体

  菊田 順一, 佐伯 行彦

  (株)南山堂, 2009年03月, 薬局, 60(3) (3), 431 - 437, 日本語


• 骨粗鬆症 脂質メディエーターを標的とした新規の骨吸収性疾患治療薬の開発

  石井 優, 菊田 順一, 佐伯 行彦, Germain Ronald N.

  (一社)日本リウマチ学会, 2009年03月, 日本リウマチ学会総会・学術集会・国際リウマチシンポジウムプログラム・抄録集, 53回・18回, 283 - 283, 日本語


• インフリキシマブ療法中、BOOPを発症したRAの2例

  田中 枝里子, 松下 正人, 菊田 順一, 石井 優, 大島 至郎, 佐伯 行彦

  (一社)日本リウマチ学会, 2009年03月, 日本リウマチ学会総会・学術集会・国際リウマチシンポジウムプログラム・抄録集, 53回・18回, 311 - 311, 日本語


• 重度の血球貪食症候群を発症したが、ステロイド・パルス療法が著効した関節リウマチの1例

  松下 正人, 菊田 順一, 田中 枝里子, 石井 優, 大島 至郎, 山中 隆夫, 原田 芳徳, 石井 泰子, 片田 圭宣, 佐伯 行彦

  (一社)日本リウマチ学会, 2009年03月, 日本リウマチ学会総会・学術集会・国際リウマチシンポジウムプログラム・抄録集, 53回・18回, 317 - 317, 日本語


• TNF阻害療法(インフリキシマブおよびエタネルセプト)無効でIL-6阻害療法が著功した難治性関節リウマチの3症例

  菊田 順一, 田中 枝里子, 石井 優, 松下 正人, 大島 至郎, 佐伯 行彦

  (一社)日本リウマチ学会, 2009年03月, 日本リウマチ学会総会・学術集会・国際リウマチシンポジウムプログラム・抄録集, 53回・18回, 389 - 389, 日本語


• 境界領域 知っておきたい インターロイキン(IL)-17

  菊田 順一, 佐伯 行彦

  (株)医学書院, 2008年10月, 臨床整形外科, 43(10) (10), 1016 - 1020, 日本語


• 低血糖発作を認め、非侵襲的にインスリノーマを否定しえたMENtype1合併の抗インスリン抗体陽性糖尿病の一例

  菊田 順一, 北村 哲宏, 遠藤 升蔵, 渡辺 長治, 佐藤 智晃, 今泉 有希, 表 順子, 大池 教子, 成瀬 亜由美, 平尾 利恵子, 幸原 晴彦

  (一社)日本糖尿病学会, 2007年04月, 糖尿病, 50(Suppl.1) (Suppl.1), S - 261, 日本語


• カルベジロールがATP感受性カリウムチャネルとG蛋白質制御カリウムチャネルのポアを阻害する

  菊田 順一, 石井 優, 岸本 健太郎, 倉智 嘉久

  (一社)日本生理学会, 2005年02月, 日本生理学雑誌, 67(2) (2), 77 - 78, 日本語


• カルベジロールがATP感受性カリウムチャネルとG蛋白質制御カリウムチャネルのポアを阻害する

  菊田 順一, 岸本 健太郎, 石井 優, 倉智 嘉久

  (公社)日本薬理学会, 2004年02月, 日本薬理学雑誌, 123(2) (2), 51P - 51P, 日本語

• 日本骨免疫学会


• 日本炎症・再生医学会


• 日本分子生物学会


• 日本臨床免疫学会


• 日本免疫学会


• 日本薬理学会


• 日本骨代謝学会


• 日本リウマチ学会


• 日本内科学会


• 日本生化学会

• バイオミネラル制御戦略に基づく新奇な結晶構造自在制御技術の開発

  丸山 美帆子, 吉川 洋史, 菊田 順一, 岡田 淳志

  日本学術振興会, 科学研究費助成事業, 挑戦的研究(開拓), 大阪大学, 2024年06月28日 - 2027年03月31日


• 多彩なマクロファージによる関節炎病態形成メカニズムの解明

  菊田 順一

  日本学術振興会, 科学研究費助成事業, 基盤研究(B), 大阪大学, 2024年04月01日 - 2027年03月31日


• 線維症の時空間的動態解析による新規治療法の開発

  菊田 順一

  国立研究開発法人科学技術振興機構, 創発的研究支援事業, 2023年04月 - 2026年03月


• 動的多細胞コミュニティの生体イメージング解析

  菊田 順一

  日本学術振興会, 科学研究費助成事業, 学術変革領域研究(B), 大阪大学, 2022年05月20日 - 2025年03月31日


• 骨イメージングではじめる動的多細胞コミュニティ学

  菊田 順一

  日本学術振興会, 科学研究費助成事業, 学術変革領域研究(B), 2022年05月 - 2025年03月


• 骨・関節破壊の病態生理の解明と新規治療法の開発

  菊田 順一

  日本学術振興会, 科学研究費助成事業, 基盤研究(B), 大阪大学, 2021年04月01日 - 2024年03月31日

  破骨細胞は、単球系血液細胞から分化・成熟する多核巨細胞で、骨破壊を担う。生理的な環境下では「骨髄」にのみ存在し、骨形成を担う骨芽細胞と協調して働くことによって骨の恒常性維持に重要な役割を果たしている（骨リモデリング）。一方、関節炎などの病的状態では、関節を包む「滑膜」を主座とする慢性炎症が、骨表面の破骨細胞の形成を促し、関節周囲の骨破壊を惹起すると言われている。これまでの破骨細胞研究の多くは、固定した骨・関節組織を切り出して顕微鏡で観察していた。この方法でも、骨が壊されている部位に多数の破骨細胞が集まっている様子は観察されたが、骨髄と炎症滑膜の二つの異なる局在・微小環境で形成された破骨細胞がどのような機能的な違いを持つのかはよく分かっていない。本研究では、これまで独自に開発・応用してきた骨髄・関節の生体二光子励起イメージング技術と、新規に開発する破骨細胞の単離・オミクス解析技術を融合させ、病的な骨・関節破壊に関わる破骨細胞の分化機構・病態生理を解析して、骨・関節破壊の本質を分子レベルで明らかにするとともに、この病的な破骨細胞を標的とした薬剤の開発の実現を目指す。本年度は、破骨細胞を特異的に蛍光標識したレポーターマウスと、破骨細胞が出す酸を感知して蛍光がONとなるpH応答性蛍光プローブを活用して、動物個体を生かしたまま生体内の骨破壊の現場をin vivoで可視化し、生理的な骨リモデリングに関わる破骨細胞と病的な骨・関節破壊に関わる破骨細胞の動態・機能を比較検討した。


• 生体イメージングによるヒト免疫細胞の動態評価系の確立

  菊田 順一

  日本学術振興会, 科学研究費助成事業, 挑戦的研究(萌芽), 大阪大学, 2021年07月09日 - 2023年03月31日

  免疫システムは、病原微生物からわれわれの体を守るために作られた、生命にとって必要不可欠な生体防御機構である。免疫学研究は、次世代シーケンサーを用いた1 細胞遺伝子発現解析など解析技術が急速に進歩し、免疫応答に関わる数多くの細胞や分子が次々と同定されている。さらに最近、生きた細胞の挙動を観察する生体イメージング研究が発展し、生体内の様々な免疫現象が明らかなってきた。一方で、マウスを中心とした疾患モデル動物で見出された多くの知見がヒトの病態の全てを反映することはできない。例えば、疾患モデルマウスではどの個体も同じように病気を発症し、薬剤に対しても画一的な反応を示すのに対して、実際の患者の症状や臨床経過、薬剤への反応性は患者によってばらつきがある。ヒトの免疫疾患の病態を理解し、より理想的な治療法を開発するためには、マウスとヒトの免疫システムの共通点と相違点を明らかにするとともに、個々のヒトの免疫応答の違いを正しく評価する必要がある。本研究では、これまで開発・応用してきた生体二光子励起イメージング技術をさらに発展させ、ヒト免疫細胞の動態・機能を解析する新たな基盤技術を確立し、ヒト疾患の病態解明と新たな治療法開発の実現を目指す。本年度は、生体内においてヒト免疫細胞が抗原を排除する機序を明らかにするため、マウスの細胞の一部をヒトの細胞に置き換えたヒト化マウスに、蛍光色素で標識したヒト免疫細胞を移植し、マウスを生かしたまま生体組織内を二光子励起顕微鏡で観察することにより、生体内におけるヒト免疫細胞の細胞動態と免疫機能を解析した。


• ４Dマルチスケールイメージング研究で解き明かす生体組織修復機構とその破綻

  菊田 順一

  国立研究開発法人日本医療研究開発機構, 革新的先端研 究開発支援事業 PRIME, 大阪大学, 2018年10月 - 2022年03月, 研究代表者


• 関節リウマチにおける炎症細胞ダイナミクスの動的解析と新規治療法の最適化

  菊田 順一

  日本学術振興会, 科学研究費助成事業, 若手研究, 大阪大学, 2019年04月01日 - 2021年03月31日

  関節リウマチ（RA）は、炎症性骨破壊を来す難治性の自己免疫疾患である。近年、様々な分子標的治療薬が関節リウマチ治療において臨床応用され、その骨破壊抑制効果が示されているが、生体内に投与した分子標的治療薬が実際に、“いつ”、“どこで”、“どのようにして”効果を発揮するのか、その薬理作用の詳細については不明な点が多い。本研究では、生体関節の二光子励起イメージング技術を駆使して、関節破壊に関わる炎症細胞の動態を可視化し関節炎の発症機序を明らかにするとともに、RA治療において臨床応用されている分子標的治療薬の生体内動態・薬効発現メカニズムを解明した。


• 蛍光生体イメージングによる細胞競合制御メカニズムの解明

  菊田 順一

  日本学術振興会, 科学研究費助成事業, 新学術領域研究(研究領域提案型), 大阪大学, 2017年04月01日 - 2019年03月31日

  近年、上皮由来がんの発生初期において、変異細胞がその周囲を正常上皮細胞に囲まれた状態で存在し、両者が生存を争う細胞競合現象が生じていることが明らかとなってきた。これまでに細胞競合に関与する細胞間シグナルや蛋白質が数多く同定されてきたが、生体内における細胞競合を制御する分子メカニズムを詳細に解明するためには、in vitroでの哺乳類培養細胞系の解析に加えて、個体を生かしたままin vivoで生きた組織内の生きた細胞を観察し、時空間的な挙動を明らかにすることが大変重要である。 本研究では、二光子励起顕微鏡を駆使して、動物個体を生かしたまま生理的な環境を維持しながら、生体腸管組織内の生きた細胞動態を可視化する生体イメージング手法を確立した。本技術を用いて、細胞競合モデルマウスの生体腸管組織内を観察し、細胞競合により変異がん細胞が正常上皮細胞層より排除される様子を経時的に可視化することに成功し、ショウジョウバエやin vitroの哺乳類上皮培養細胞系で明らかになった細胞競合現象が、哺乳類生体内のin vivoの系でも実際に起こっていることが明らかとなった。さらに、正常上皮細胞や変異細胞の動きや形態変化など得られたイメージング画像を定量的に数理解析するとともに、生理学的・病理学的ながん微小環境を維持した上で細胞競合の際に起こる多彩な遺伝子発現変化を1細胞レベルで定量的に解析する新たな方法論を構築した。本研究で確立した技術は、今後、細胞競合制御メカニズムの解明において強力なツールになると考えられる。


• 蛍光生体イメージングで読み解くウイルス共生とその破綻

  菊田 順一

  日本学術振興会, 科学研究費助成事業, 新学術領域研究(研究領域提案型), 大阪大学, 2017年04月01日 - 2019年03月31日

  長い進化の歴史の中で、ウイルスとヒトはお互いに適応しながら共生してきた。一方で、ヒトの体の中には多種多様な免疫細胞が存在し、日々ウイルスと闘いを繰り広げている。「生体内でウイルスが実際にどのように振る舞ってヒトの細胞と共生するのか」、「ヒトの免疫細胞はウイルスにどのような攻撃をして共生関係を破綻させるのか」など、ウイルスとヒトの共生の“実態”を理解するためには、“生きた”組織内で“生きた”ウイルスや細胞を観察し、時空間的な挙動を明らかにすることが大変重要である。 近年、マウスの細胞の一部をヒトの細胞に置き換えたヒト化マウスモデルが確立し、ヒトに共生するウイルスのin vivo解析が可能となったが、フローサイトメトリーやELISAなどの従来の解析手法では、生体内における動態をリアルタイムで評価することはできなかった。一方、蛍光生体イメージング技術が発展し、個体を生かしたまま生体内の宿主細胞とウイルスの挙動を可視化できるようになったが、そのいずれもがマウスを中心としたモデル動物を使用していたため、ウイルスがヒトの細胞に及ぼす影響を解析することは困難であった。 本研究では、“ヒト化マウス技術”と“蛍光生体イメージング技術”を融合させることで、ヒト化マウスの生体組織内においてウイルスの挙動を可視化することに成功した。さらに、ヒト抗ウイルス免疫応答の制御メカニズムを明らかにするため、ウイルスとヒト免疫細胞の相互作用を可視化し、その細胞動態（細胞の動きや形態変化）を定量解析した。本研究で確立した技術は、今後、ウイルスとヒト細胞の共生の実態の解明において強力なツールになると考えられる。


• 蛍光生体イメージング技術による関節リウマチの病態解明と新規薬効評価系の開発

  菊田 順一

  日本学術振興会, 科学研究費助成事業, 若手研究(A), 大阪大学, 2015年04月01日 - 2018年03月31日

  関節リウマチは、炎症性骨破壊を来す難治性の自己免疫疾患である。近年、様々な生物学的製剤が関節リウマチ治療において臨床応用され、その骨破壊抑制効果が示されているが、生体内における各種薬剤の作用機序の差異についてはよく分かっていない。本研究では、骨・関節破壊の現場をin vivoで可視化する蛍光生体イメージング系を確立し、炎症性骨破壊における生きた細胞の動態を明らかにした。さらに、本技術を用いて、各種生物学的製剤が炎症によって誘導された破骨細胞に及ぼす効果を解析し、薬剤間の薬効の差異を解明した。


• 蛍光生体イメージング技術による免疫細胞の骨吸収制御機構の解明

  菊田 順一

  日本学術振興会, 科学研究費助成事業, 新学術領域研究(研究領域提案型), 大阪大学, 2015年04月01日 - 2017年03月31日

  骨組織は、骨格の維持だけでなく、血液系幹細胞から多種多様な免疫細胞が生成される造血の場であり、免疫システム構築の場でもある。骨髄腔内では血管が縦横無尽に走行し、その血管壁には多くの開窓が存在するという特徴を持つ。骨髄腔で生成された免疫細胞は、この開窓を利用して骨髄腔と血管腔を出入りし、末梢に遊走する、もしくは末梢より骨髄に戻る事で、免疫系を維持していると考えられている。生体内で感染や炎症が生じると、多くの免疫細胞が骨髄腔から炎症部位に動員され、細胞遊走が活発になる。骨髄内の血管透過性によって制御された免疫系の細胞遊走システムは、生体の恒常性維持だけでなく、生体防御反応においても必要不可欠な機能である。精緻に制御された免疫システムの本質を理解するためには、骨髄腔内を流れる豊富な循環血流を保ったまま、生体骨組織内の細胞動態を四次元で解析することが重要である。 本研究では、これまで独自に開発・応用してきた骨の蛍光生体イメージング技術を活用して、細胞集団の相互的空間情報や生理活性物質の濃度など生理的な骨髄環境を保持したまま、骨髄腔内の免疫細胞動態を四次元で可視化し1細胞レベルで機能解析する技術開発を行った。本研究で確立した生体二光子励起イメージング技術は、様々な免疫細胞の遊走や機能を1細胞レベルで解析することができるため、今後、免疫細胞の遊走システムの本質を明らかにし、免疫系の細胞遊走システムが担う生体恒常性維持機構の解明にもつながると考えられる。


• 蛍光生体イメージング技術を駆使した細胞競合現象の解明

  菊田 順一

  日本学術振興会, 科学研究費助成事業, 新学術領域研究(研究領域提案型), 大阪大学, 2015年04月01日 - 2017年03月31日

  近年、上皮由来がんの発生初期において、変異細胞がその周囲を正常上皮細胞に囲まれた状態で存在し、両者が生存を争う細胞競合現象が生じていることが明らかとなってきた。これまでに細胞競合に関与する細胞間シグナルや蛋白質が数多く同定されてきたが、生体内における細胞競合を制御する分子メカニズムを詳細に解明するためには、in vitroでの哺乳類培養細胞系の解析に加えて、個体を生かしたままin vivoで生きた組織内の生きた細胞を観察し、時空間的な挙動を明らかにすることが大変重要である。 本研究では、二光子励起顕微鏡を駆使して、動物個体を生かしたまま生理的な環境を維持しながら、生体腸管組織内の生きた細胞動態を可視化する生体イメージング手法を確立した。さらに、確立した生体二光子励起イメージング技術を用いて、細胞競合モデルマウス（Villin-CreERT2; CAG-LSL-RasV12-IRES-EGFP）の生体腸管組織内を観察し、細胞競合によりRas変異がん細胞が正常上皮細胞層より排除される様子をリアルタイムで可視化することに成功した。一方、Ras変異が誘導されないコントロールマウス（Villin-CreERT2; CAG-LSL-IRES-EGFP）では、EGFP陽性細胞は腸上皮層より排除されなかった。これらの結果から、ショウジョウバエやin vitroの哺乳類上皮培養細胞系で明らかになった細胞競合現象が、哺乳類生体内のin vivoの系でも実際に起こっている現象であることが明らかとなった。


• 生体多光子励起イメージングによる関節リウマチの病態生理の解明

  菊田 順一

  日本学術振興会, 科学研究費助成事業, 研究活動スタート支援, 大阪大学, 2013年08月30日 - 2015年03月31日

  本研究では、関節リウマチの病態生理を解明するために、生体多光子励起顕微鏡を駆使して関節炎・関節破壊の現場をin vivoで可視化するライブイメージング系を確立し、炎症関節内における生きた破骨細胞の動態を明らかにした。さらに、関節リウマチ治療において臨床応用されている分子標的治療薬が破骨細胞の骨吸収に及ぼす効果をin vivoで明らかにした。本研究で確立した生体イメージング技術は、今後、関節リウマチの発症メカニズムの解明だけでなく、新たな関節リウマチ治療薬の開発においても強力な手段となることが強く期待される。


• 生体多光子励起イメージングによる関節炎・関節破壊の機序解明

  菊田 順一

  日本学術振興会, 科学研究費助成事業, 特別研究員奨励費, 大阪大学, 2012年 - 2013年, 研究代表者

  関節リウマチは、日本の人口の約0.5%の患者を有する最も頻度の高い自己免疫疾患の一つであり、滑膜の炎症に伴う進行性の骨破壊を生じるため、患者の運動機能が著しく制限される。そのため、骨破壊による関節機能障害の制御が、リウマチ治療の最大の課題である。関節リウマチにおける関節破壊は、関節を包む滑膜の炎症から始まり、滑膜の増殖、パンヌスの形成、骨・軟骨破壊へと進行していく。その病態形成には、破骨細胞、マクロファージ、T細胞が複雑に関与すると考えられている。しかしながら、「それぞれの細胞が、いつどのようにして関節内に遊走してきて、関節炎が発症するのか」という関節破壊に関わる細胞の遊走制御や動態については、これまでほとんど解明されていない。 そこで、本研究初年度では、関節炎・関節破壊の現場をin vivoで可視化するべく技術開発を行い、生体多光子励起イメージング系を独自に改良することで、動物の生きた関節内における生きた破骨細胞やマクロファージの遊走を経時的に観察することができる実験系を確立した。さらに、関節リウマチ治療薬の薬効評価を単一細胞レベルでリアルタイムに行うことも可能となった。本研究で確立した関節内ライブイメージング系は、今後、関節リウマチの関節炎・関節破壊における「細胞遊走」のメカニズムの解明や、「細胞遊走」を標的とした新たな関節リウマチ治療法の開発において強力な手段となり得ると考えられる。

  競争的資金

• 関節炎治療剤

  石井 優, 菊田 順一, 長谷川 哲雄

  JP2019043057, 2019年11月01日, 国立大学法人大阪大学, WO2020-091052, 2020年05月07日

  特許権

  外部リンク


• 評価方法、評価装置およびプログラム

  石井 優, 菊田 順一, 松田 秀雄, 瀬尾 茂人

  特願2018-007571, 2018年01月19日, 国立大学法人大阪大学, 特開2019-128148, 2019年08月01日

  特許権

  外部リンク


• 画像処理装置および方法、並びにプログラム

  米谷 信彦, 石井 優, 菊田 順一

  特願2011-232479, 2011年10月24日, 株式会社ニコン, 国立大学法人大阪大学, 特開2013-085546, 2013年05月13日

  特許権

  外部リンク