SEARCH

Search Details

AOI Michiyo

Researcher basic information

■ Research Keyword
  • 幹細胞
  • 再生医学
  • 核初期化
  • 分化多能性
  • 転写因子
  • microRNA
  • ES細胞
  • iPS細胞
  • 体細胞の初期化
  • レトロウイルス
■ Research Areas
  • Life sciences / Medical biochemistry
  • Life sciences / Virology

Research activity information

■ Paper
  • Tomohiro Miyamoto, Naomasa Fukase, Teruya Kawamoto, Shuichi Fujiwara, Hitomi Hara, Ryoko Sawada, Yuta Nakamatsu, Yutaka Mifune, Kenichiro Kakutani, Yuichi Hoshino, Shinya Hayashi, Tomoyuki Matsumoto, Takehiko Matsushita, Michiyo Koyanagi-Aoi, Takashi Aoi, Toshiyuki Takemori, Shunsuke Yahiro, Ryosuke Kuroda, Toshihiro Akisue
    Cancer stem cells (CSCs) have been implicated as critical mediators in the progression, chemoresistance and metastatic capabilities of diverse malignancies, including osteosarcoma (OS). The authors have succeeded in generating CSC‑like cells (MG‑OKS) from the OS cell line MG‑63 by transducing defined factors. A significant increase in small proline‑rich protein 1A (SPRR1A) expression, a cross‑linked envelope protein in keratinocytes, was observed in MG‑OKS cells. Therefore, SPRR1A could be involved in tumor initiation, growth and poor OS progression. However, its specific role in OS remains unclear. The present study aimed to evaluate the role of SPRR1A in OS both in vitro and in vivo using MG‑OKS cells. Three experimental groups were established: MG‑OKS cells transfected with SPRR1A small interfering (si)RNA (siMG‑OKS), untransfected MG‑OKS cells and MG‑OKS cells transfected with scrambled siRNA (scMG‑OKS) as controls. SPRR1A expression, morphological changes, cell proliferation and migration were assessed in these groups. RNA sequencing was performed to examine the genetic changes caused by SPRR1A suppression. To evaluate tumorigenicity in vivo, cells from each group were subcutaneously transplanted into the backs of nude mice. Tumor volume and Ki‑67 expression were assessed and compared among the three groups at four weeks post‑transplantation. The siMG‑OKS group exhibited altered cell morphology, reduced cell proliferation and decreased migratory abilities in vitro. RNA sequencing revealed suppression of genes involved in cell adhesion in the siMG‑OKS group. Furthermore, the in vivo tumorigenicity of siMG‑OKS was lower than that of the other two experimental groups. These findings suggest that SPRR1A is one of the key cell adhesion‑related molecules involved in OS progression, potentially serving as a therapeutic target for this refractory tumor. However, further research is needed to fully elucidate the mechanisms by which SPRR1A influences OS pathogenesis and to explore its clinical potential.
    Feb. 2025, Oncology reports, 53(2) (2), English, International magazine
    Scientific journal
    Link to Kobe Univ. Repository (Kernel)

  • Katsuya Sato, Michiyo Koyanagi-Aoi, Keiichiro Uehara, Yosuke Yamashita, Masakazu Shinohara, Suji Lee, Anika Reinhardt, Knut Woltjen, Koji Chiba, Hideaki Miyake, Masato Fujisawa, Takashi Aoi
    Late-onset hypogonadism (LOH) syndrome is characterized by age-related testosterone deficiency and negatively affects the quality of life of older men. A promising therapeutic approach for LOH syndrome is transplantation of testosterone-producing Leydig-like cells (LLCs) derived from human induced pluripotent stem cells (hiPSCs). However, previous studies have encountered obstacles, such as limited cell longevity, insufficient testosterone production, and inefficiency of differentiation. To address these issues, we developed a novel protocol that includes forced NR5A1 expression, a cytokine cocktail promoting mesoderm differentiation, and a transitional shift from 3D to 2D cultures. The resultant cells survived on culture dishes for over 16 weeks, produced 22-fold more testosterone than the conventional method, and constituted a homogeneous population of LLCs with a differentiation efficiency exceeding 99% without purification. Furthermore, these LLCs were successfully engrafted subcutaneously into mice, resulting in increased serum testosterone levels. Our study will facilitate innovative therapeutic strategies for LOH syndrome.
    Jan. 2025, Stem cell reports, 102392 - 102392, English, International magazine
    Scientific journal
    Link to Kobe Univ. Repository (Kernel)

  • 宮本 智弘, 藤原 周一, 原 仁美, 深瀬 直政, 澤田 良子, 中松 裕太, 青井 貴之, 小柳 三千代, 黒田 良祐, 秋末 敏宏
    (公社)日本整形外科学会, Sep. 2024, 日本整形外科学会雑誌, 98(8) (8), S1967 - S1967, Japanese

  • Erika Tanaka, Michiyo Koyanagi-Aoi, So Nakagawa, Sayumi Shimode, Hideto Yamada, Yoshito Terai, Takashi Aoi
    Elsevier BV, Jun. 2024, Regenerative Therapy, 26, 729 - 740
    Scientific journal
    Link to Kobe Univ. Repository (Kernel)

  • Chihiro Takemori, Michiyo Koyanagi-Aoi, Takeshi Fukumoto, Makoto Kunisada, Kazumasa Wakamatsu, Shosuke Ito, Chieko Hosaka, Seiji Takeuchi, Akiharu Kubo, Takashi Aoi, Chikako Nishigori
    Elsevier BV, Jun. 2024, Journal of Dermatological Science
    Scientific journal
    Link to Kobe Univ. Repository (Kernel)

  • LOH症候群に対する新しい再生医療の開発
    佐藤 克哉, 山下 遥介, 河村 駿, 賀來 泰大, 千葉 公嗣, 小柳 三千代, 青井 貴之, 藤澤 正人
    (一社)日本性機能学会, Aug. 2023, 日本性機能学会雑誌, 38(2) (2), 131 - 131, Japanese

  • 宮本 智弘, 藤原 周一, 原 仁美, 深瀬 直政, 澤田 良子, 八尋 俊輔, 青井 貴之, 小柳 三千代, 黒田 良祐, 秋末 敏宏
    (公社)日本整形外科学会, Aug. 2023, 日本整形外科学会雑誌, 97(8) (8), S1892 - S1892, Japanese

  • 男性医学2023 テストステロンを増やすための基礎研究から生活習慣まで ヒトiPS細胞由来Leydig細胞の作製
    青井 貴之, 佐藤 克哉, 小柳 三千代
    (一社)日本抗加齢医学会, Jun. 2023, 日本抗加齢医学会総会プログラム・抄録集, 23回, 125 - 125, Japanese

  • Kohei Yamakawa, Michiyo Koyanagi-Aoi, Akihito Machinaga, Nobuyuki Kakiuchi, Tomonori Hirano, Yuzo Kodama, Takashi Aoi
    Abstract Background Our study and several studies have reported that in some cancers, including pancreatic ductal adenocarcinoma (PDAC), the expression of squamous lineage markers, such as esophagus-tissue-specific genes, correlated with a poor prognosis. However, the mechanism by which the acquisition of squamous lineage phenotypes leads to a poor prognosis remains unclear. We previously reported that retinoic acid signaling via retinoic acid receptor γ (RARγ signaling) determines the differentiation lineage into the esophageal squamous epithelium. These findings hypothesized that the activation of RARγ signaling contributed to acquiring squamous lineage phenotypes and malignant behavior in PDAC. Methods This study utilized public databases and immunostaining of surgical specimens to examine RARγ expression in PDAC. We evaluated the function of RARγ signaling by inhibitors and siRNA knockdown using a PDAC cell line and patient-derived PDAC organoids. The mechanism of the tumor-suppressive effects by blocking RARγ signaling was examined by a cell cycle analysis, apoptosis assays, RNA sequencing and Western blotting. Results RARγ expression in pancreatic intraepithelial neoplasia (PanIN) and PDAC was higher than that in the normal pancreatic duct. Its expression correlated with a poor patient prognosis in PDAC. In PDAC cell lines, blockade of RARγ signaling suppressed cell proliferation by inducing cell cycle arrest in the G1 phase without causing apoptosis. We demonstrated that blocking RARγ signaling upregulated p21 and p27 and downregulated many cell cycle genes, including cyclin-dependent kinase 2 (CDK2), CDK4 and CDK6. Furthermore, using patient-derived PDAC organoids, we confirmed the tumor-suppressive effect of RARγ inhibition and indicated the synergistic effects of RARγ inhibition with gemcitabine. Conclusions This study clarified the function of RARγ signaling in PDAC progression and demonstrated the tumor-suppressive effect of selective blockade of RARγ signaling against PDAC. These results suggest that RARγ signaling might be a new therapeutic target for PDAC.
    Springer Science and Business Media LLC, May 2023, Cancer Cell International, 23(1) (1)
    Scientific journal
    Link to Kobe Univ. Repository (Kernel)

  • Nobuyuki Murai, Michiyo Koyanagi-Aoi, Hiroto Terashi, Takashi Aoi
    Elsevier BV, Mar. 2023, Stem Cell Reports
    Scientific journal
    Link to Kobe Univ. Repository (Kernel)

  • Kenji Miura, Michiyo Koyanagi-Aoi, Yoshimasa Maniwa, Takashi Aoi
    Abstract Background The chorioallantoic membrane (CAM) assay is a well-established technique to evaluate tumor invasion and angiogenesis and may overcome the shortcoming of the patient-derived xenograft (PDX) mouse model. Currently, few reports have described lung cancer invasion and angiogenesis in the CAM assay. We therefore used the CAM assay in the evaluation of lung cancer. Method Lung cancer cell line-derived organoids or lung cancer cell lines were transplanted into the CAM on embryonic development day (EDD) 10, and an analysis was performed on EDD 15. Microscopic and macroscopic images and movies of the grafts on the CAM were captured and analyzed. The relationships between the graft and chick vessels were evaluated using immunohistochemistry. Results We transplanted lung cancer cell lines and cell line-derived organoid into a CAM to investigate angiogenesis and invasion. They engrafted on the CAM at a rate of 50–83%. A549-OKS cells showed enhanced cell invasion and angiogenesis on the CAM in comparison to A549-GFP cells as was reported in vitro. Next, we found that A549-TIPARP cells promoted angiogenesis on the CAM. RNA-seq identified 203 genes that were upregulated more than twofold in comparison to A549-GFP cells. A pathway analysis revealed many upregulated pathways related to degradation and synthesis of the extracellular matrix in A549-TIPARP cells. Conclusions The CAM assay can be used to evaluate and research invasion and angiogenesis in lung cancer. The elevated expression of TIPARP in lung cancer may induce angiogenesis by remodeling the extracellular matrix.
    Springer Science and Business Media LLC, Feb. 2023, Cancer Cell International, 23(1) (1)
    Scientific journal
    Link to Kobe Univ. Repository (Kernel)

  • Shunsuke Yahiro, Teruya Kawamoto, Shuichi Fujiwara, Hitomi Hara, Naomasa Fukase, Ryoko Sawada, Toshiyuki Takemori, Tomohiro Miyamoto, Yutaka Mifune, Kenichiro Kakutani, Yuichi Hoshino, Shinya Hayashi, Tomoyuki Matsumoto, Takehiko Matsushita, Michiyo Koyanagi-Aoi, Takashi Aoi, Ryosuke Kuroda, Toshihiro Akisue
    Ewing sarcoma (ES) is an aggressive primary malignant bone tumor that predominantly affects children and young adults. Multimodal treatment approaches have markedly improved the survival of patients with localized ES. However, local recurrence and distant metastasis following curative therapies remain a main concern for patients with ES. Recent studies have suggested that slow‑cycling cells (SCCs) are associated with tumor progression, local recurrence and distant metastasis in various types of cancers. According to the results of these studies, it was hypothesized that SCCs may play a critical role in tumor progression, chemoresistance and local/distal recurrence in patients with ES. The present study applied a label‑retaining system using carboxyfluorescein diacetate succinimidyl ester (CFSE) to identify and isolate SCCs in ES cell lines. In addition, the properties of SCCs, including sphere formation ability, cell cycle distribution and chemoresistance, in comparison with non‑SCCs were investigated. RNA sequencing also revealed several upregulated genes in SCCs as compared with non‑SCCs; the identified genes not only inhibited cell cycle progression, but also promoted the malignant properties of SCCs. On the whole, the present study successfully identified SCCs in ES cells through a label‑retaining system using CFSE. Moreover, to the best of our knowledge, the present study is the first to describe the characteristic properties of SCCs in ES. The findings of this study, if confirmed, may prove to be useful in elucidating the underlying molecular mechanisms and identifying effective therapeutic targets for ES.
    Nov. 2022, International journal of oncology, 61(5) (5), English, International magazine
    Scientific journal
    Link to Kobe Univ. Repository (Kernel)

  • Kohei Yamakawa, Michiyo Koyanagi-Aoi, Keiichiro Uehara, Atsuhiro Masuda, Hiroaki Yanagimoto, Hirochika Toyama, Takumi Fukumoto, Yuzo Kodama, Takashi Aoi
    Objectives Small proline-rich protein 1A (SPRR1A) is recognized as a squamous differentiation marker but is also upregulated in some non-squamous cancers. However, its expression in pancreatic ductal adenocarcinoma (PDAC) has not been investigated. This study elucidated the expression of SPRR1A in PDAC and its effect on the prognosis and malignant behavior of PDAC. Methods We examined the SPRR1A expression by immunohistochemistry in 86 surgical PDAC cases and revealed the relationship between its expression and the prognosis of the PDAC patients. Furthermore, we overexpressed SPRR1A in pancreatic cancer cell lines (PK-1 and Panc-1) and assessed the phenotype and gene expression changes in vitro. Results Among the 84 cases, excluding 2 with squamous differentiation, 31 (36.9%) had a high SPRR1A expression. The overall survival (median 22.1 months vs. 33.6 months, p = 0.0357) and recurrence-free survival (median 10.7 months vs. 15.5 months, p = 0.0298) were significantly lower in the high-SPRR1A-expression group than in the low-SPRR1A-expression group. A multivariate analysis indicated that a high SPRR1A expression (HR 1.706, 95% CI 1.018 to 2.862, p = 0.0427) and residual tumor status (HR 2.687, 95% CI 1.487 to 4.855, p = 0.00106) were independent prognostic factors. The analysis of TCGA transcriptome data demonstrated that the high-SPRR1A-expression group had a significantly worse prognosis than the low-SPRR1A-expression group, which supported our data. SPRR1A overexpression in PK-1 and Panc-1 did not result in remarkable changes to in vitro phenotypes, such as the cell proliferation, chemo-resistance, EMT, migration or global gene expression. Conclusion Increased expression of SPRR1A is associated with a poor prognosis in PDAC and may serve as a novel prognostic marker. However, our in vitro study suggests that the SPRR1A expression may be a consequence, not a cause, of the aggressive behavior of PDAC.
    Public Library of Science (PLoS), May 2022, PLOS ONE, 17(5) (5), e0266620 - e0266620, English, International magazine
    Scientific journal
    Link to Kobe Univ. Repository (Kernel)

  • Takahiro Koide, Michiyo Koyanagi-Aoi, Keiichiro Uehara, Yoshihiro Kakeji, Takashi Aoi
    Intestinal metaplasia is related to gastric carcinogenesis. Previous studies have suggested the important role of CDX2 in intestinal metaplasia, and several reports have shown that the overexpression of CDX2 in mouse gastric mucosa caused intestinal metaplasia. However, no study has examined the induction of intestinal metaplasia using human gastric mucosa. In the present study, to produce an intestinal metaplasia model in human gastric mucosa in vitro, we differentiated human-induced pluripotent stem cells (hiPSC) to gastric organoids, followed by the overexpression of CDX2 using a tet-on system. The overexpression of CDX2 induced, although not completely, intestinal phenotypes and the enhanced expression of many, but not all, intestinal genes and previously reported intestinal metaplasia-related genes in the gastric organoids. This model can help clarify the mechanisms underlying intestinal metaplasia and carcinogenesis in human gastric mucosa and develop therapies to restitute precursor conditions of gastric cancer to normal mucosa.
    Elsevier BV, May 2022, iScience, 25(5) (5), 104314 - 104314, English, International magazine
    Scientific journal
    Link to Kobe Univ. Repository (Kernel)

  • 小寺澤 康文, 小柳 三千代, 押切 太郎, 掛地 吉弘, 青井 貴之
    (NPO)日本気管食道科学会, Apr. 2022, 日本気管食道科学会会報, 73(2) (2), 165 - 165, Japanese

  • Keiichiro Uehara, Michiyo Koyanagi-Aoi, Takahiro Koide, Tomoo Itoh, Takashi Aoi
    Human gastric development has not been well studied. The generation of human pluripotent stem cell-derived gastric organoids (hGOs) comprising gastric marker-expressing epithelium without an apparent smooth muscle (SM) structure has been reported. We modified previously reported protocols to generate hGOs with muscularis mucosa (MM) from hiPSCs. Time course analyses revealed that epithelium development occurred prior to MM formation. Sonic hedgehog (SHH) and TGF-β1 were secreted by the epithelium. HH and TGF-β signal inhibition prevented subepithelial MM formation. A mechanical property of the substrate promoted SM differentiation around hGOs in the presence of TGF-β. TGF-β signaling was shown to influence the HH signaling and mechanical properties. In addition, clinical specimen findings suggested the involvement of TGF-β signaling in MM formation in recovering gastric ulcers. HH and TGF-β signaling from the epithelium to the stroma and the mechanical properties of the subepithelial environment may influence the emergence of MM in human stomach tissue.
    Elsevier BV, Feb. 2022, Stem Cell Reports, 17(4) (4), 820 - 834, English, International magazine
    Scientific journal
    Link to Kobe Univ. Repository (Kernel)

  • Takaki Ishida, Michiyo Koyanagi-Aoi, Daisuke Yamamiya, Atsushi Onishi, Katsuya Sato, Keiichiro Uehara, Masato Fujisawa, Takashi Aoi
    Abstract Late-onset hypogonadism (LOH) syndrome, due to a partial lack of testosterone, decreases the quality of life of older men. Testosterone is mainly secreted by Leydig cells in the testes. Leydig cell transplantation is expected to be a promising alternative to conventional testosterone replacement therapy for LOH syndrome. We herein report a simple and robust protocol for directed differentiation of human induced pluripotent stem cells (hiPSCs) into Leydig-like cells by doxycycline-inducible overexpression of NR5A1 and treatment with a combination of 8-bromoadenosine-3′,5′-cyclic monophosphate (8-Br-cAMP) and forskolin. The differentiated cells expressed the steroidogenic enzyme genes STAR, CYP11A1, CYP17A1, and HSD3B2 and the specific markers of adult Leydig cells HSD17B3, INSL3, and LHCGR. Furthermore, we confirmed the secretion of functional testosterone from the cells into the culture supernatant by a testosterone-sensitive cell proliferation assay. These findings showed that the hiPSCs were able to be differentiated into Leydig-like cells, supporting the expectation that hiPSC-derived Leydig-like cells can be novel tools for treating LOH syndrome.
    The Endocrine Society, Dec. 2021, Endocrinology, 162(12) (12)
    Scientific journal
    Link to Kobe Univ. Repository (Kernel)

  • Mariko Taniguchi-Ikeda, Michiyo Koyanagi-Aoi, Tatsuo Maruyama, Toru Takaori, Akiko Hosoya, Hiroyuki Tezuka, Shotaro Nagase, Takuma Ishihara, Taisuke Kadoshima, Keiko Muguruma, Keiko Ishigaki, Hidetoshi Sakurai, Akira Mizoguchi, Bennett G. Novitch, Tatsushi Toda, Momoko Watanabe, Takashi Aoi
    Elsevier BV, Oct. 2021, iScience, 24(10) (10), 103140 - 103140
    Scientific journal
    Link to Kobe Univ. Repository (Kernel)

  • Shuichi Fujiwara, Teruya Kawamoto, Yohei Kawakami, Yasufumi Koterazawa, Hitomi Hara, Toshiyuki Takemori, Kazumichi Kitayama, Shunsuke Yahiro, Kenichiro Kakutani, Tomoyuki Matsumoto, Takehiko Matsushita, Takahiro Niikura, Michiyo Koyanagi-Aoi, Takashi Aoi, Ryosuke Kuroda, Toshihiro Akisue
    BACKGROUND: Cancer stem cells (CSCs) are considered to be responsible for tumor initiation, formation, and poor prognosis of cancer patients. However, the rarity of CSCs in clinical samples makes it difficult to elucidate characteristics of CSCs, especially in osteosarcoma (OS). The aim of this study is to verify whether it is possible to generate CSC-like cells by transducing defined factors into an OS cell line. METHODS: We retrovirally transduced the Octamer-binding transcription factor 3/4 (OCT3/4), Kruppel-like factor 4 (KLF4), and SRY-box transcription factor 2 (SOX2) genes into the MG-63 human OS cell line (MG-OKS). Parental and GFP-transduced MG-63 cells were used as negative control. We assessed the properties of the generated cells in vitro and in vivo. Multiple comparisons among groups were made using a one-way analysis of variance (ANOVA) followed by post hoc testing with Tukey's procedure. RESULTS: MG-OKS cells in vitro exhibited the significantly increased mRNA expression levels of CSC markers (CD24, CD26, and CD133), decreased cell growth, increased chemoresistance and cell migration, and enhanced sphere formation. Notably, MG-OKS cells cultured under osteogenic differentiation conditions showed strongly positive staining for both Alizarin Red S and alkaline phosphatase, indicating osteogenesis of the cells. Gene ontology analysis of microarray data revealed significant upregulation of epidermal-related genes. Tumors derived from MG-OKS cells in vivo were significantly larger than those from other cells in μCT analysis, and immunohistochemical staining showed that Ki-67, osteocalcin, and HIF-1α-positive cells were more frequently detected in the MG-OKS-derived tumors. CONCLUSIONS: In this study, we successfully generated OS CSC-like cells with significantly enhanced CSC properties following transduction of defined factors.
    Oct. 2020, Stem cell research & therapy, 11(1) (1), 429 - 429, English, International magazine
    Scientific journal
    Link to Kobe Univ. Repository (Kernel)

  • Yasufumi Koterazawa, Michiyo Koyanagi-Aoi, Keiichiro Uehara, Yoshihiro Kakeji, Takashi Aoi
    In the original publication of the article, the following errors were noted and corrected in this correction.
    Oct. 2020, Journal of gastroenterology, 55(10) (10), 1010 - 1011, English, Domestic magazine

  • Yasufumi Koterazawa, Michiyo Koyanagi-Aoi, Keiichiro Uehara, Yoshihiro Kakeji, Takashi Aoi
    BACKGROUND: The esophagus is known to be derived from the foregut. However, the mechanisms regulating this process remain unclear. In particular, the details of the human esophagus itself have been poorly researched. In this decade, studies using human induced pluripotent stem cells (hiPSCs) have proven powerful tools for clarifying the developmental biology of various human organs. Several studies using hiPSCs have demonstrated that retinoic acid (RA) signaling promotes the differentiation of foregut into tissues such as lung and pancreas. However, the effect of RA signaling on the differentiation of foregut into esophagus remains unclear. METHODS: We established a novel stepwise protocol with transwell culture and an air-liquid interface system for esophageal epithelial cell (EEC) differentiation from hiPSCs. We then evaluated the effect of all-trans retinoic acid (ATRA), which is a retinoic acid receptor (RAR)α, RARβ and RARγ agonist, on the differentiation from the hiPSC-derived foregut. Finally, to identify which RAR subtype was involved in the differentiation, we used synthetic agonists and antagonists of RARα and RARγ, which are known to be expressed in esophagus. RESULTS: We successfully generated stratified layers of cells expressing EEC marker genes that were positive for lugol staining. The enhancing effect of ATRA on EEC differentiation was clearly demonstrated with quantitative reverse transcription polymerase chain reaction, immunohistology, lugol-staining and RNA sequencing analyses. RARγ agonist and antagonist enhanced and suppressed EEC differentiation, respectively. RARα agonist had no effect on the differentiation. CONCLUSION: We revealed that RARγ activation promotes the differentiation of hiPSCs-derived foregut into EECs.
    Jun. 2020, Journal of gastroenterology, 55(8) (8), 763 - 774, English, Domestic magazine
    [Refereed]
    Scientific journal
    Link to Kobe Univ. Repository (Kernel)

  • Chieko Hosaka, Makoto Kunisada, Michiyo Koyanagi-Aoi, Taro Masaki, Chihiro Takemori, Mariko Taniguchi-Ikeda, Takashi Aoi, Chikako Nishigori
    Induced pluripotent stem cell (iPSC) technology offers a novel approach for conversion of human primary fibroblasts into melanocytes. During attempts to explore various protocols for differentiation of iPSCs into melanocytes, we found a distinct and self-renewing cell lineage that could differentiate into melanocytes, named as melanocyte precursor cells (MPCs). The MPCs exhibited a morphology distinctive from that of melanocytes, in lacking either the melanosomal structure or the melanocyte-specific marker genes MITF, TYR, and SOX10. In addition, gene expression studies in the MPCs showed high-level expression of WNT5A, ROR2, which are non-canonical WNT pathway markers, and its related receptor TGFβR2. In contrast, MPC differentiation into melanocytes was achieved by activating the canonical WNT pathway using the GSK3β inhibitor. Our data demonstrated the distinct characteristic of MPCs' ability to differentiate into melanocytes, and the underlying mechanism of interfacing between canonical WNT signaling pathway and non-canonical WNT signaling pathway.
    Sep. 2019, Pigment cell & melanoma research, 32(5) (5), 623 - 633, English, International magazine
    [Refereed]
    Scientific journal
    Link to Kobe Univ. Repository (Kernel)

  • Kotaro Suzuki, Michiyo Koyanagi-Aoi, Keiichiro Uehara, Nobuyuki Hinata, Masato Fujisawa, Takashi Aoi
    For augmentation or reconstruction of urinary bladder after cystectomy, bladder urothelium derived from human induced pluripotent stem cells (hiPSCs) has recently received focus. However, previous studies have only shown the emergence of cells expressing some urothelial markers among derivatives of hiPSCs, and no report has demonstrated the stratified structure, which is a particularly important attribute of the barrier function of mature bladder urothelium. In present study, we developed a method for the directed differentiation of hiPSCs into mature stratified bladder urothelium. The caudal hindgut, from which the bladder urothelium develops, was predominantly induced via the high-dose administration of CHIR99021 during definitive endoderm induction, and this treatment subsequently increased the expressions of uroplakins. Terminal differentiation, characterized by the increased expression of uroplakins, CK13, and CK20, was induced with the combination of Troglitazone + PD153035. FGF10 enhanced the expression of uroplakins and the stratification of the epithelium, and the transwell culture system further enhanced such stratification. Furthermore, the barrier function of our urothelium was demonstrated by a permeability assay using FITC-dextran. According to an immunohistological analysis, the stratified uroplakin II-positive epithelium was observed in the transwells. This method might be useful in the field of regenerative medicine of the bladder.
    Jul. 2019, Scientific reports, 9(1) (1), 10506 - 10506, English, International magazine
    [Refereed]
    Scientific journal
    Link to Kobe Univ. Repository (Kernel)

  • Kotaro Suzuki, Michiyo Koyanagi-Aoi, Keiichiro Uehara, Nobuyuki Hinata, Takashi Aoi, Masato Fujisawa
    Apr. 2019, Journal of Urology, English
    [Refereed]
    Scientific journal

  • Kotaro Suzuki, Michiyo Koyanagi-Aoi, Keiichiro Uehara, Nobuyuki Hinata, Takashi Aoi, Masato Fujisawa
    Springer Science and Business Media {LLC}, Mar. 2019, European Urology Supplements, 18(1) (1), e1763, English
    [Refereed]
    Scientific journal

  • Daisuke Watanabe, Michiyo Koyanagi-Aoi, Mariko Taniguchi-Ikeda, Yukiko Yoshida, Takeshi Azuma, Takashi Aoi
    John Wiley and Sons Ltd., Jan. 2018, Stem Cells Translational Medicine, 7(1) (1), 34 - 44, English
    [Refereed]
    Scientific journal
    Link to Kobe Univ. Repository (Kernel)

  • Ryo Ishida, Michiyo Koyanagi-Aoi, Nobu Oshima, Yoshihiro Kakeji, Takashi Aoi
    Oct. 2017, MOLECULAR CANCER RESEARCH, 15(10) (10), 1455 - 1466, English
    [Refereed]
    Scientific journal

  • Hiroyuki Ogawa, Michiyo Koyanagi-Aoi, Kyoko Otani, Yoh Zen, Yoshimasa Maniwa, Takashi Aoi
    Sep. 2017, SCIENTIFIC REPORTS, 7(1) (1), 12317, English
    [Refereed]
    Scientific journal
    Link to Kobe Univ. Repository (Kernel)

  • Masatoshi Nishizawa, Kazuhisa Chonabayashi, Masaki Nomura, Azusa Tanaka, Masahiro Nakamura, Azusa Inagaki, Misato Nishikawa, Ikue Takei, Akiko Oishi, Koji Tanabe, Mari Ohnuki, Hidaka Yokota, Michiyo Koyanagi-Aoi, Keisuke Okita, Akira Watanabe, Akifumi Takaori-Kondo, Shinya Yamanaka, Yoshinori Yoshida
    Sep. 2016, CELL STEM CELL, 19(3) (3), 341 - 354, English
    [Refereed]
    Scientific journal

  • Bernhard Payer, Michael Rosenberg, Masashi Yamaji, Yukihiro Yabuta, Michiyo Koyanagi-Aoi, Katsuhiko Hayashi, Shinya Yamanaka, Mitinori Saitou, Jeannie T. Lee
    Dec. 2013, Molecular Cell, 52(6) (6), 805 - 818, English, Co-authored internationally
    [Refereed]
    Scientific journal

  • Michiyo Koyanagi-Aoi, Mari Ohnuki, Kazutoshi Takahashi, Keisuke Okita, Hisashi Noma, Yuka Sawamura, Ito Teramoto, Megumi Narita, Yoshiko Sato, Tomoko Ichisaka, Naoki Amano, Akira Watanabe, Asuka Morizane, Yasuhiro Yamada, Tosiya Sato, Jun Takahashi, Shinya Yamanaka
    Dec. 2013, PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 110(51) (51), 20569 - 20574, English
    [Refereed]
    Scientific journal

  • Kyoko Miura, Yohei Okada, Takashi Aoi, Aki Okada, Kazutoshi Takahashi, Keisuke Okita, Masato Nakagawa, Michiyo Koyanagi, Koji Tanabe, Mari Ohnuki, Daisuke Ogawa, Eiji Ikeda, Hideyuki Okano, Shinya Yamanaka
    Aug. 2009, NATURE BIOTECHNOLOGY, 27(8) (8), 743 - 745, English
    [Refereed]
    Scientific journal

  • Masato Nakagawa, Michiyo Koyanagi, Koji Tanabe, Kazutoshi Takahashi, Tomoko Ichisaka, Takashi Aoi, Keisuke Okita, Yuji Mochiduki, Nanako Takizawa, Shinya Yamanaka
    Jan. 2008, NATURE BIOTECHNOLOGY, 26(1) (1), 101 - 106, English
    [Refereed]
    Scientific journal

  • M Koyanagi, M Hijikata, K Watashi, O Masui, K Shimotohno
    Apr. 2005, JOURNAL OF BIOLOGICAL CHEMISTRY, 280(13) (13), 12430 - 12437, English
    [Refereed]
    Scientific journal

  • RelA suppresses the Wnt/beta-catenin pathway without exerting trans-acting transcriptional ability
    O Masui, Y Ueda, A Tsumura, M Koyanagi, M Hijikata, K Shimotohno
    May 2002, INTERNATIONAL JOURNAL OF MOLECULAR MEDICINE, 9(5) (5), 489 - 493, English
    [Refereed]
    Scientific journal

■ MISC
  • 下垂体発生における視床下部-口腔外胚葉間FGFの役割
    松本 隆作, 青井 貴之, 小柳 三千代, 須賀 英隆, 福岡 秀規, 井口 元三, 小川 渉, 高橋 裕
    (一社)日本内分泌学会, Apr. 2019, 日本内分泌学会雑誌, 95(1) (1), 408 - 408, Japanese

  • 下垂体発生における視床下部-口腔外胚葉間FGFの役割
    松本 隆作, 青井 貴之, 小柳 三千代, 須賀 英隆, 福岡 秀規, 井口 元三, 小川 渉, 高橋 裕
    (一社)日本内分泌学会, Apr. 2019, 日本内分泌学会雑誌, 95(1) (1), 352 - 352, Japanese

  • 誘導性多能性幹細胞の神経分化と治療的効果(Neural differentiation and therapeutic effects of induced pluripotent stem cell)
    三浦 恭子, 辻 収彦, 岡田 洋平, 西野 誠, 富里 周太, 幸田 和久, 池田 栄二, 沖田 圭介, 高橋 和利, 小柳 三千代, 田邊 剛士, 柚崎 通介, 戸山 芳昭, 中村 雅也, 山中 伸弥, 岡野 栄之
    (公社)日本生化学会, Nov. 2008, 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, 81回・31回, 1T25 - 10, English

■ Books And Other Publications
  • ヒト内在性レトロウイルスと疾患
    青井(小柳)三千代, 青井貴之
    医学のあゆみ 第273巻12号, Jun. 2020

  • ヒト多能性幹細胞株の分化特性とばらつき
    青井(小柳)三千代, 青井貴之
    日本臨床73巻増刊号5, p74-79, 2015

  • 多能性幹細胞の標準化に向けて
    青井(小柳)三千代, 山中伸弥
    ファルマシア vol.48, No.9, p872-874, 2012

  • 多能性幹細胞とmicroRNA
    小柳三千代, 山中伸弥
    生体の科学vol.61, No.4, p332-337, 2010

  • 多能性幹細胞とmicroRNA
    小柳三千代, 山中伸弥
    遺伝子医学MOOK 最新RNAと疾患研究 p186-191, 2009

  • iPS細胞の樹立と今後の展望
    小柳三千代, 山中伸弥
    臨床検査 53巻, 1号, p123-128, 2009

  • がん遺伝子Mycを用いない人工多能性幹細胞の作成
    小柳三千代, 中川誠人, 山中伸弥
    BIO INDUSTRY 4号, p89-95, 2008

  • マウス線維芽細胞培養から誘導される多能性幹細胞
    小柳三千代, 山中伸弥
    遺伝子医学MOOK 進み続ける細胞移植治療の実際（上巻）p124-127, 2008

■ Lectures, oral presentations, etc.
  • 多能性幹細胞を⽤いた消化器領域の発生・病態研究
    青井貴之, 小柳三千代
    第23回日本再生医療学会総会, Mar. 2024

  • 大腸がんで発現するHERV由来タンパク質の局在と機能の解析
    青井(小柳)三千代, 中川 草, 青井 貴之
    第46回日本分子生物学会年会, Dec. 2023

  • Establishment of PHOX2B-GFP reporter iPS cell lines derived from healthy individuals and congenital central hypoventilation syndrome patients
    Takumi Kido, Yukito Imagawa, Michiyo Koyanagi, Takashi Aoi, Kazumichi Fujioka
    The 22nd Congress of the Federation of Asia and Oceania Perinatal Societies, Oct. 2023

  • 福山型筋ジストロフィーに対する糖鎖増強療法
    池田 真理子, 小柳 三千代, 森田 健太, 櫻井 英俊, 原田 陽一郎, 丸山 達生, 青井 貴之
    第9回日本筋学会学術集会, Aug. 2023

  • 患者由来 iPS 細胞を用いた先天性中枢性低換気症候群の病態メカニズムの解析
    城戸拓海, 小柳三千代, 藤岡一路, 青井貴之
    第22回日本再生医療学会総会, Mar. 2023

  • GENERATION OF HUMAN IPS CELL-DERIVED LEYDIG CELLS AND ENCAPSULATION IN IMPLANTABLE DEVICES
    Katsuya Sato, Michiyo Koyanagi-Aoi, Takashi Aoi
    ISSCR 2022 Annual Meeting (San Francisco), Jun. 2022

  • iPS細胞を用い細胞老化に着目した色素性乾皮症の病態解明
    福本毅, 竹森千尋, 錦織千佳子, 久保亮治, 青井(小柳)三千代, 青井貴之
    第121回日本皮膚科学会総会, Jun. 2022

  • 先天性中枢性低換気症候群患者由来iPS細胞の樹立と神経細胞への分化誘導
    城戸拓海, 小柳三千代, 藤岡一路, 荒田晶子, 青井貴之
    第21回日本再生医療学会総会, Mar. 2022

  • ヒトiPS細胞由来Leydig細胞の作製
    佐藤克哉, 大西篤史, 賀來泰大, 石田貴樹, 岡田桂輔, 千葉公嗣, 藤澤正人, 小柳三千代, 青井貴之
    第109回 日本泌尿器科学会総会, Dec. 2021, Japanese

  • Epigenetic regulation in melanocytes differentiated from induced pruripotent stem cells originated from xeroderma pigmentosum
    Chihiro Takemori, Takeshi Fukumoto, Michiyo Koyanagi-Aoi, Makoto Kunisada, Chieko Hosaka, Takashi Aoi, Chikako Nishigori
    第46回日本研究皮膚科学会学術大会, English

  • ヒトiPS細胞由来Leydig細胞の作製法の改良と，分化過程の解明
    佐藤克哉, 大西篤史, 賀來泰大, 石田貴樹, 岡田桂輔, 千葉公嗣, 松下経, 藤澤正人, 小柳三千代, 青井貴之
    第66回 日本生殖医学会学術講演会・総会, Nov. 2021, Japanese

  • 患者由来大腸癌オルガノイドを用いたバルプロ酸の新規治療薬としての有用性の検討
    堀江 和正, 堀川 学, 小柳 三千代, 青井 貴之, 松田 武, 掛地 吉弘
    第63回日本消化器病学会大会, Japanese

  • Generation of human iPS cell-derived Leydig cells
    Katsuya Sato, Atsushi Onishi, Yasuhiro Kaku, Takaki Ishida, Keisuke Okada, Koji Chiba, Kei Matsusita, Michiyo Koyanagi-Aoi, Masato Fujisawa, Takashi Aoi
    第73回 西日本泌尿器科学会総会, Nov. 2021, English

  • Generation of human iPS cell-derived Leydig cells
    Katsuya Sato, Michiyo Koyanagi-Aoi, Takashi Aoi
    ISSCR/JSRM 2021 Tokyo International Symposium, Oct. 2021, English

  • The epithelial-derived factors and mechanical environment induce the muscularis mucosa of the iPSC-derived human gastric organoid
    Keiichiro Uehara, Michiyo Koyanagi-Aoi, Takashi Aoi
    ISSCR/JSRM 2021 Tokyo International Symposium, Oct. 2021, English

  • Restoration of the defect in radial glial fiver cell migration and cortical plare organization in brain organoid model of Fukyama mascular dystrophy
    Taniguchi-Ikeda M, Koyanagi-Aoi M, Muguruma K, Sakurai H, Novitch B, Watanabe M, Aoi T
    ISSCR/JSRM 2021 Tokyo International Symposium, English

  • Usefulness of Melanocytes Differentiated from Induced Pluripotent Stem Cells for Research on Pathology of Xeroderma Pigmentosum
    Chihiro Takemori, Michiyo Koyanagi-Aoi, Takeshi Fukumoto, Makoto Kunisada, Chieko Hosaka, Takashi Aoi, Chikako Nishigori
    ISSCR/JSRM 2021 Tokyo International Symposium, English

  • Re-generation of cytotoxic γδT cells from human γδT-derived iPSCs
    Nobuyuki Murai, Michiyo Koyanagi-Aoi, Hiroto Terashi, Takashi Aoi
    ISSCR/JSRM 2021 Tokyo International Symposium, English

  • Valproic acid suppresses the tissue-reconstructing ability of colorectal cancer stem cells by inhibition of GSK3
    Kazumasa Horie, Manabu Horikawa, Michiyo Koyanagi-Aoi, Takashi Aoi
    ISSCR/JSRM 2021 Tokyo International Symposium, English

  • 色素性乾皮症A群由来iPS細胞より分化したメラノサイトを用いた網羅的遺伝子発現解析
    竹森 千尋, 福本 毅, 青井(小柳)三千代, 国定 充, 保坂 千恵子, 青井 貴之, 錦織 千佳子
    第30回日本色素細胞学会学術大会, Oct. 2021, Japanese

  • Ewing肉腫におけるslow-cycling cellsの同定
    八尋 俊輔, 河本 旭哉, 原 仁美, 竹森 俊幸, 藤原 周一, 北山 和道, 宮本 智弘, 青井 貴之, 青井 三千代, 黒田 良祐, 秋末 敏宏
    第36回日本整形外科学会基礎学術集会, Japanese

  • ヒトiPS細胞由来Leydig細胞の作製法の改良と，分化過程の解明
    佐藤克哉, 大西篤史, 賀來泰大, 石田貴樹, 岡田桂輔, 千葉公嗣, 松下経, 藤澤正人, 小柳三千代, 青井貴之
    第71回 日本泌尿器科学会中部総会, Oct. 2021, Japanese

  • Comprehensive analyses on melanocytes differentiated from induced pruripotent stem cells originated from xeroderma pigmentosum complementation group A
    Chihiro Takemori, Michiyo Koyanagi-Aoi, Takeshi Fukumoto, Makoto Kunisada, Chieko Hosaka, Takashi Aoi, Chikako Nishigori
    The 50th European Society for Dermatological Research Annual Meeting, English

  • Ewing肉腫におけるslow cycling cellsの同定
    八尋 俊輔, 河本 旭哉, 川上 洋平, 原 仁美, 竹森 俊幸, 藤原 周一, 北山 和道, 宮本 智弘, 青井 貴之, 青井(小柳)三千代, 黒田 良祐, 秋末 敏宏
    第54回日本整形外科学会骨・軟部腫瘍学術集会, Japanese

  • 上皮由来SHH、TGFβシグナル及び基質の硬度がhiPSC由来胃オルガノイドの粘膜筋板を誘導する
    上原慶一郎, 小柳三千代, 伊藤智雄, 青井貴之
    第110回日本病理学会総会, Japanese

  • ヒトiPS細胞由来Leydig細胞の作製
    佐藤克哉, 大西篤史, 賀來泰大, 石田貴樹, 岡田桂輔, 千葉公嗣, 松下経, 藤澤正人, 小柳三千代, 青井貴之
    第18回 泌尿器科再建再生研究会, Jun. 2021, Japanese

  • 上皮由来SHH、TGFβシグナル及び気質の硬度がhiPSC由来胃オルガノイドの粘膜筋板を誘導する
    上原慶一郎, 小柳三千代, 伊藤智雄, 青井貴之
    第20回日本再生医療学会総会, Japanese

  • γδT由来iPS細胞から再誘導したγδT細胞の腫瘍傷害性
    村井信幸, 青井貴之, 小柳三千代, 寺師浩人
    第20回日本再生医療学会総会, Japanese

  • 人工肺癌幹細胞を用いた、肺癌組織再構築と新規治療法開発の試み
    Ogawa Hiroyuki, Aoi Michiyo, Aoi Takashi, Maniwa Yoshimasa
    第58回日本肺癌学会学術集会, Oct. 2017, Japanese, 日本肺癌学会, 横浜, Domestic conference
    Oral presentation

  • 人工大腸がん幹細胞を用いたがん幹細胞の特性解析とがん治療への応用
    Aoi Michiyo
    東海大学医学部セミナー, Sep. 2017, Japanese, Tokai University, 小田原, Domestic conference
    Public discourse

  • 人工癌幹細胞と癌オルガノイドを用いた癌幹細胞研究
    Aoi Takashi, 石田 諒, Ogawa Hiroyuki, Aoi Michiyo, Kakeji Yoshihiro, Maniwa Yoshimasa
    第27回日本サイトメトリー学会学術集会, Jun. 2017, Japanese, 日本サイトメトリー学会, 神戸, Domestic conference
    [Invited]
    Nominated symposium

  • 人工肺癌幹細胞による、肺癌組織再構築の試み
    Ogawa Hiroyuki, Shimizu Nahoko, Hokka Daisuke, Tanaka Yugo, Aoi Michiyo, Aoi Takashi, Maniwa Yoshimasa
    第34回日本呼吸器外科学会総会, May 2017, Japanese, 日本呼吸器外科学会, 福岡, Domestic conference
    Oral presentation

  • Interleukin-6 blockade, a novel cancer stem cell targeted therapy, attenuates lung cancer tissue construction.
    Ogawa Hiroyuki, Aoi Michiyo, Aoi Takashi, Maniwa Yoshimasa
    American Association for Cancer Research annual meeting 2017, Apr. 2017, English, American Association for Cancer Research, Washington D.C., USA, International conference
    Poster presentation

■ Affiliated Academic Society
  • THE MOLECULAR BIOLOGY SOCIETY OF JAPAN

  • 日本ウイルス学会

■ Research Themes
  • B型肝炎ウイルスの生活環と病原性の機序解明による革新的創薬基盤研究
    勝二 郁夫, 西辻 裕紀, 立花太郎, 阿部 隆之, Hussein Hassan Aly, 青井(小柳)三千代
    国立研究開発法人日本医療研究開発機構, Ｂ型肝炎創薬実用化等研究事業, Apr. 2025 - Mar. 2028, Coinvestigator

  • 福山型筋ジストロフィー及び類縁疾患に対する低分子化合物Mn007を用いた糖鎖増強療法の実用化にむけた研究
    池田真理子, 青井貴之, 小柳三千代, 丸山達生, 原田陽一郎, 犬塚 達俊
    AMED難治性疾患実用化研究事業, Apr. 2024 - Mar. 2027

  • ヒト内在性レトロウイルス様配列由来Gag類似タンパク質の大腸がんにおける 機能解析
    青井(小柳)三千代
    基盤研究C, Apr. 2024 - Mar. 2027

  • 大腸癌に対する腫瘍免疫活性を増強するエピジェネティクス標的薬の探索と作用機構解明
    青井貴之、掛地吉弘、青井(小柳)三千代
    基盤研究B, Apr. 2022 - Mar. 2026

  • FCMDのαジストログリカン糖鎖のホメオスタシスに着目した治療法開発
    池田真理子, 青井貴之, 石垣景子, 中嶋和紀, 丸山達生, 青井(小柳)三千代, 長坂美和子
    基盤研究B, Apr. 2021 - Mar. 2026

  • レポーターHBVを駆使したB型肝炎ウイルス増殖機構の解析と創薬 ターゲットの探索・同定に資する研究
    勝二 郁夫, 下遠野 邦忠, 西辻 裕紀, 立花太郎, 三木大樹, 杉山 真也, 宮川敬, Hussein Hassan Aly, 阿部 隆之, 伊藤 昌彦, 上田 優輝, 青井(小柳)三千代
    AMED肝炎等克服実用化研究事業, Apr. 2022 - Mar. 2025

  • FCMD及び類縁疾患のiPSCs由来三次元培養法による疾患モデルを駆使した 病態評価と低分子治療法開発
    池田真理子, 青井(小柳)三千代, 森田健太, 原田陽一郎
    AMED疾患特異的iPS細胞の利活用促進・難病研究加速 プログラム, Jun. 2021 - Mar. 2024

  • 疾患特異的iPS細胞を用いた先天性中枢性低換気症候群における低CO2感受性の 分子機構
    藤岡一路, 山本暢之, 青井(小柳)三千代, 荒田晶子, 上坂敏弘
    AMED疾患特異的iPS細胞の利活用促進・難病研究加速 プログラム, Jun. 2021 - Mar. 2024

  • Functional analysis of HERV in colorectal cancer
    青井(小柳)三千代 中川 草
    Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C), Grant-in-Aid for Scientific Research (C), Kobe University, Apr. 2019 - Mar. 2022

  • Identification of prognostic factors and development of preclinical POC for novel therapies for colorectal cancer based on molecular mechanisms of cancer stem cells.
    Takashi Aoi
    Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B), Grant-in-Aid for Scientific Research (B), Kobe University, Apr. 2018 - Mar. 2022
    In this study, we tried to obtain preclinical POC for a novel treatment of colorectal cancer based on the molecular mechanism of cancer stem cells. We established a system to generate organoids derived from human clinical specimens of colorectal cancer and to quantitatively evaluate the efficacy of drugs using these organoids, as well as a system to create xenografts by transplanting these organoids into immunodeficient mice and to evaluate the efficacy of drugs against these xenografts. Using these systems, we examined the efficacy of the novel therapeutic drug candidates that the applicants had discovered in our previous studies using colon cancer cell lines, and revealed that they were effective in the systems.

  • 福山型筋ジストロフィーに対する低分子化合物スクリーニングを用いた分子標的治療法開発
    池田真理子;青井貴之;丸山達生
    国立研究開発法人日本医療研究開発機構, 日本医療研究開発機構研究費, Apr. 2018 - Mar. 2021, Coinvestigator

  • アセンブラーとしての癌/非癌幹細胞の機能解明とその制御技術の開発
    青井貴之, 蓮沼誠久, 荒木通啓, 山本一彦, 青井三千代
    AMED幹細胞・再生医学イノベーション創出プログラム, Nov. 2016 - Mar. 2019

  • Aoi Takashi, HOTTA Haku
    Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C), Grant-in-Aid for Scientific Research (C), Kobe University, Apr. 2014 - Mar. 2017
    In the process of cancer initiation and development, escape from tumor immunosurveillance has been thought to be important. However its molecular mechanisms are still unclear. This study aimed to clarify regulatory mechanisms of the expression of immune target molecules induced by viral infection of human non-cancer hepatocytes by utilizing iPS cell technologies and establish a system for drug discovery targeting it. In this study, we established a drug-inducible system for forced expression of each proteins encoded by hepatitis C virus in human iPS cell - derived hepatocytes. Using this system, we analyzed the regulatory mechanism of the expression of immune target molecules.
    Competitive research funding

  • 青井 三千代
    学術研究助成基金助成金／若手研究(B), Apr. 2014 - Mar. 2016
    Competitive research funding

  • 体細胞核の初期化に関わるmicroRNAの機能解析
    小柳 三千代
    日本学術振興会, 科学研究費助成事業, 特別研究員奨励費, 京都大学, 2009 - 2011
    ヒト胚性幹(ES)細胞やヒトiPS細胞は様々な細胞へと分化することができる一方で、クローン間で分化傾向に違いがあることも報告されている。我々は様々な年齢、性別のドナーの様々な組織から、いろいろな方法で49個のiPS細胞を樹立した。また、これらのiPS細胞のmicroRNAの発現、遺伝子発現、DNAのメチル化を10株のES細胞と比較した。その結果、ES細胞とiPS細胞は分子レベルで非常に似通っており、両者を完全に区別できるようなマーカーは存在しなかった。次に、このような分子レベルでのばらつきと、神経細胞への分化傾向との関連を調べた。ES/iPS細胞をSFEBq法で分化させ、分化誘導開始後14日目に初期の神経分化マーカーPSA-NCAMと未分化細胞のマーカーOCT3/4の発現をフローサイトメトリーで調べた。ほとんどのクローンが、90%以上の割合でPSA-NCAM陽性の神経細胞へと分化したが、ごくまれに未分化細胞マーカーであるOCT3/4が10%程度残っているクローンが存在した。我々は後者のクローンを"bad"クローンと定義づけ、一方で、実験を繰り返しても未分化細胞の残存率が1%を超えない細胞を"good"クローンと定義した。分化誘導を行う前に、"bad"クローンと"good"クローンが予測できるかどうかを調べるために、我々は"bad"クローンと"good"クローンの未分化状態での遺伝子発現とmiRNA発現をマイクロアレイで比較した。その結果、いくつかの分子が"bad"クローンで高く発現している傾向にあることがわかった。これらの因子を組み合わせてマーカーとして用いることができれば、臨床応用に適さない、移植後に未分化腫瘍を形成する可能性のあるクローンを分化誘導前に除去することができると期待される。現在は、これらの因子の制御機構について解析し、報告する準備をしている。

  • Molecular mechanisms underlying nuclear reprogramming of somatic cells
    YAMANAKA Shinya, AOI Takashi, NAKAGAWA Masato, TAKAHASHI Kazutoshi, OKITA Keisuke, YOSHIDA Yoshinori, WATANABE Akira, YAMAMOTO Takuya, KNUT Woltjen, KOYANAGI Michiyo
    Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Specially Promoted Research, Grant-in-Aid for Specially Promoted Research, Kyoto University, 2007 - 2011
    iPS cells can be generated by introduction of four reprogramming factors into somatic cells. We have found that chimera mice produced with c-Myc show high tumorigenicity. To overcome this issue, we could generate iPS cells without Myc under the modified conditions. But Myc minus iPS cells showed less pluripotency compared to iPS cells generated with c-Myc. Next, we explored the factors which could replace the c-Myc function. We found L-Myc as a candidate. We could confirm that L-Myc enhances the efficiency of iPS generation and that the tumorigenicity of L-Myc is hardly observed. So, we can obtain the safer iPS cells using L-Myc. We established the system to examine the safety of iPS cells using the iPS cells-derived neuronal cells. We also established the differentiation methods of iPS cells into hepatocytes, blood cells, and cardiomyocytes in vitro. Next generation sequencer(NGS) is very powerful tool for analysis of genetic properties of iPS cells. We have analyzed the gene expression, DNA methylation, changes of splicing pattern in iPS cells using NGS

  • Functional analysis of microRNAs which are involved in reprogramming of somatic cells
    KOYANAGI Michiyo
    Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research, Grant-in-Aid for Young Scientists (B), Kyoto University, 2007 - 2008
    マウス、ヒトの線維芽細胞にOct3/4, Sox2, c-Myc,Klf4の4つの因子を導入することによって様々な細胞へと分化できる能力を持ったES細胞と同様の性質を持つ細胞(iPS細胞)ができる。この時、がん遺伝子Mycを導入しなくてもiPS細胞を作製できること、また、Mycを除くとMycを含む4因子で導入した場合と比較してiPS細胞の誘導効率は減少するが、その際に、特定のmicroRNAをレトロウイルスを使って共発現させることによって、iPS細胞の誘導効率が上昇することがわかった。

■ Industrial Property Rights
  • 色素幹細胞の製造方法
    錦織千佳子, 青井貴之, 保坂千恵子, 国定充, 青井三千代
    特願2015-242724, 11 Dec. 2015, 特表2017-104085, 15 Jun. 2017
    Patent right

  • 大腸がん幹細胞の維持増殖方法、及び大腸がんオルガノイドの誘導方法
    青井貴之, 石田諒, 青井三千代, 掛地吉弘
    特願2017-110626, 05 Jun. 2017
    Patent right

  • 人工多能性幹細胞の選別方法
    山中伸弥, 高橋和利, 小柳三千代, 大貫茉里
    特願2014-503890, 25 Jul. 2012, 特表523735A, 18 Sep. 2014, 特許6143265
    Patent right

  • 人工多能性幹細胞の選別方法
    山中伸弥, 小柳三千代
    特願2012-548681, 17 Jan. 2011, 特表2013-516982, 16 May 2013, 特許5936131
    Patent right

  • 効率的な核初期化方法
    山中伸弥, 小柳三千代
    特願2008-335129, 26 Dec. 2008, 特開2010-158171, 22 Jul. 2010, 特許5626619
    Patent right

  • 効率的な核初期化法
    山中伸弥, 小柳三千代
    PCT/JP2008/059586, 23 May 2008, WO2009/075119, 18 Jun. 2009, 特許5558097
    Patent right

TOP