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増見 恭子大学院科学技術イノベーション研究科 科学技術イノベーション専攻准教授
研究活動情報
■ 受賞- 2017年09月 化学工学会, バイオ部会優秀ポスター賞, 昆虫細胞による2Aペプチドを用いた抗体タンパク質の生産国内学会・会議・シンポジウム等の賞
- 2005年06月 日本乳癌学会, 研究奨励賞, 「網羅的遺伝子発現解析による乳癌のドセタキセル効果予測診断法の開発」
- 2005年03月 大阪対がん協会, 研究奨励賞, 「網羅的遺伝子発現解析による乳癌のドセタキセル効果予測診断法の開発」
- 2002年03月 大阪対がん協会, 研究奨励賞, 「乳癌の予後を決定する遺伝子群の同定」
- Recombinant adeno-associated viruses (rAAVs) are useful vectors for clinical gene therapy. It is crucial to examine DNA impurities, such as plasmid DNA, for quality control of rAAV products. In this study, we examined highly purified rAAV1, rAAV2, rAAV5, and rAAV6 samples produced on a three-plasmid platform, using a high-throughput sequencer. These samples contained 0.49–3.80% detectable DNA impurities derived from the three plasmids, as estimated by the ampicillin resistance gene ( amp R ). The plasmid impurities consisted of 90.62–95.84% pAAV, 3.21–6.83% pRC, and 0.95–2.55% pHelper DNA. These trends were consistent with those of DNA impurities determined by droplet digital PCR (ddPCR), indicating that cleaved pAAV backbone DNA was the primary source of DNA impurities encapsulated into the capsids. To examine the preferential encapsulation of pAAV backbone DNA into capsids, short sequencing reads were mapped to the entire pAAV backbone sequence, and we found that the reads were relatively evenly distributed across the backbone sequence, with occasional sharp drops. Furthermore, the read length distribution containing the pAAV backbone sequence showed a main peak at 3.2 kb in Oxford Nanopore Technologies sequencing. This length was consistent with that of backbone DNA nicked at two terminal resolution sites by Rep78/68. Analysis of the terminal sequences of the long reads containing backbone sequences revealed that 92.7–98.4% of them contained Rep-binding elements. These results indicate that the pAAV backbone in linear DNA form was cleaved from pAAV-ZsGreen1 by Rep78/68 nicking at trs in the nucleus, and that the single-stranded DNA was efficiently translocated into the capsids through the encapsulating machinery, similar to the rAAV genome.SAGE Publications, 2026年02月, Human Gene Therapy, 37(7-8) (7-8), 349 - 364[査読有り]研究論文（学術雑誌）
- Recombinant adeno-associated virus (rAAV) vectors are among the most effective for gene therapy. A significant advancement in rAAV vector production is developing the triple-plasmid transfection method, which remains the most widely used technique. In this study, we used Expi293FTM (Expi293F) and Viral Production Cells 2.0 (VPC2.0 cells) to evaluate various transfection reagents, comparing transgene protein expression levels and intracellular plasmid copy numbers to optimize rAAV production. Our findings indicated that the effectiveness of transfection reagents in promoting higher rAAV production was cell-dependent and that rAAV productivity correlated more with plasmid levels in the cell nucleus than with transgene protein expression levels. Confocal laser microscopy revealed that in cells transfected with the high-yield transfection reagent, a large amount of free plasmid DNA entered the nucleus, whereas the transfection reagents themselves did not. These results provide new insights into the intracellular mechanisms underlying efficient rAAV vector production. Furthermore, identifying transfection reagents that facilitate nuclear plasmid delivery will aid in the selection of optimal reagents for high-yield AAV production.2026年01月, Human gene therapy, 37(1-2) (1-2), 56 - 64, 英語, 国際誌[査読有り]研究論文（学術雑誌）
- INTRODUCTION: Safe and effective adeno-associated virus (AAV) vectors are essential for gene therapy. Particle heterogeneity, specifically particle DNA of varying types and sizes, significantly affects recombinant adeno-associated virus (rAAV) performance. Previous studies have identified particle-associated DNAs; however, the specific DNA composition of these mixed populations remains poorly understood. This study aimed to investigate the DNA composition of the isolated subpopulations of rAAV particles obtained through anion exchange (AEX) chromatography. METHODS: RAAV2-ZsGreen1 particles were fractionated on an AEX column, resulting in 12 distinct fractions. We analyzed the DNA composition of these fractions using droplet digital PCR (ddPCR) and MiSeq to identify the incorporated DNA heterogeneity in them. RESULTS: Our findings illustrated a clear trend in which the DNA content increasing in fractions was associated with an increased rAAV genomic DNA ratio of total particle DNA. The particle DNA content increased significantly across fractions from Peak 1 to Peak 2, showing approximately 30,000- and 5000-fold increases for ZsGreen1 (rAAV genome) and ampR (plasmid impurity), respectively. Notably, in the empty particle subpopulations, the rate of detectable DNA molecules was lower than one DNA fragment per 100 particles, with inverted terminal repeat (ITR) sequences being the most prevalent. CONCLUSIONS: With the elucidated profile of particle DNAs, this study provides detailed information on particle heterogeneity, shedding light on empty and partial particle formation and impurity DNA incorporation.2025年06月, Biotechnology journal, 20(6) (6), e70031, 英語, 国際誌[査読有り]研究論文（学術雑誌）
- Recombinant adeno-associated viruses (rAAVs) are widely used viral vectors in human gene therapy. However, DNA impurities, such as plasmid DNA and host cell DNA, remain a significant quality control concern for final products. Our study examined purified rAAV1-ZsGreen1, rAAV2-ZsGreen1, rAAV5-ZsGreen1, and rAAV6-ZsGreen1 samples and found that they contained 0.69-3.27% DNA impurities derived from three plasmids, as detected by droplet digital PCR. These plasmid-derived impurities primarily consisted of those derived from the pAAV plasmid (≥98.88%), with small amounts of pRC1, pRC2mi342, pRC5, or pRC6 (≤0.91%), and pHelper (≤0.21%) plasmids. To determine the DNA strand form of these impurities within the capsids, we used two different DNases with distinct substrate specificities. The extracted DNA impurities from the rAAV samples exhibited high sensitivity to nuclease P1 but not to lambda exonuclease. Similarly, host cell DNA encapsulated within the capsids revealed similar sensitivities to the nucleases. These findings indicate that DNA impurities derived from the plasmids and host cell DNA are encapsulated into rAAV capsids as single-stranded DNA, likely through a mechanism similar to that of the rAAV genome.2025年05月, Human gene therapy, 36(9-10) (9-10), 814 - 822, 英語, 国際誌[査読有り]研究論文（学術雑誌）
- AMED iD3 Catalyst Unit, 2025年, Translational and Regulatory Sciences, 7(2) (2), 57 - 62[査読有り]研究論文（学術雑誌）
- Parvoviruses have evolved to possess a linear single-stranded DNA (ssDNA) genome ranging from 4 to 6.3 kb. Adeno-associated virus (AAV), a member of the Parvoviridae family, contains approximately 5 kb of linear ssDNA within its capsid. This ssDNA features two 145-base inverted terminal repeats (ITRs) positioned at each end. ITRs have a T-shaped hairpin secondary structure, which plays a crucial role in viral replication. To investigate the impact of ITRs on ssDNA stability, we conducted a DNA denaturation-reannealing assay in 10 mM magnesium acetate, 50 mM potassium acetate, and 20 mM Tris-acetate buffer at pH7.9. Conventional double-stranded DNA (dsDNA) fragments retain a reannealing capability of over 50% for sizes under 8.8 kb, gradually losing this capability as sizes increase; however, dsDNA fragments in rAAV ranging from 0.7 to 6.3 kb did not exhibit a reannealing profile. This suggests that the presence of ITRs at both ends hinders annealing between the complementary strands. These results indicate that ITR structures preferentially induce an ssDNA conformation less than 6.3 kb in size, and that the stability of AAV ssDNA contributes to the viral life cycle, including processes such as infection, replication, and packaging. Considering the size of parvovirus genomes, it appears that their genomes require reversible flexibility in complementary DNA strands; simultaneously, this adaptability needs to be regulated by specific palindromic ITRs at both ends.2024年11月, Scientific reports, 14(1) (1), 27696 - 27696, 英語, 国際誌[査読有り]研究論文（学術雑誌）
- Recombinant adeno-associated virus (rAAV) is a prominent viral vector currently available for human gene therapy. The diameter of the rAAV capsid is ∼25 nm, and a positive or negative single-stranded DNA is packaged within the vector capsid. In this report, we describe a concise method to examine the extracted rAAV genome using an automated electrophoresis system. The rAAV genome, prepared from vector particles through either heat treatment at 95°C for 10 min or the phenol-chloroform extraction method, was analyzed using an automated electrophoresis system under denaturation conditions. The heat treatment protocol demonstrated a comparable yield with the phenol-chloroform extraction protocol, and the quantified amounts of the rAAV genome obtained using the automated electrophoresis system were consistent with those quantitated by quantitative PCR. Additionally, crude rAAV extractions could also be analyzed by the automated electrophoresis system after DNase I treatment. These results indicated that this simple and quick analysis using automated electrophoresis is highly useful for confirming the purity and integrity of the rAAV genome.2024年02月, Human gene therapy, 35(3-4) (3-4), 104 - 113, 英語, 国際誌[査読有り]研究論文（学術雑誌）
- Adeno-associated virus (AAV) vectors are produced as a mixture of the desired particle (full particle, FP), which is filled with the designed DNA, product-related impurities such as particle without DNA (empty particle, EP), and aggregates. Cesium chloride or iodixanol equilibrium density gradient ultracentrifugation (DGE-UC) has been used for the purification of AAV vectors. DGE-UC can separate FP from impurities based on the difference in their buoyant densities. Here, we report the applications and limitations of equilibrium density gradient analytical ultracentrifugation (DGE-AUC) using a modern AUC instrument that employs DGE-UC principles for the characterization and quantitation of AAV vectors. We evaluated the quantitative ability of DGE-AUC in comparison with sedimentation velocity AUC (SV-AUC) or band sedimentation AUC (BS-AUC) using AAVs with different DNA lengths and different serotypes. DGE-AUC enabled the accurate quantification of the ratio of FP to EP when the AAV vector primarily contains these particles. Furthermore, we developed a new workflow to identify the components of separated peaks in addition to FP and EP. Ultraviolet absorption spectra obtained by multiwavelength detection can also support peak assignment following component identification. DGE-AUC experiments for AAV vectors have limitations with regard to minor components with low absorption at the detected wavelength or those with a density similar to that of major components of AAV vectors. DGE-AUC is the only analytical method that can evaluate particle density heterogeneity; therefore, SV-AUC or BS-AUC and DGE-AUC are complementary methods for reliable assessment of the purity of AAV vectors.2024年01月, Analytical chemistry, 96(2) (2), 642 - 651, 英語, 国際誌[査読有り]研究論文（学術雑誌）
- Abstract The recombinant adeno‐associated viral (rAAV) vector is one of the most effective viral vectors in gene therapy because of its low immunogenicity, high transduction efficiency, broad tissue specificity, and long‐term transgene expression ability. HEK293T cells are widely used to produce rAAV vectors, and acquisition of high rAAV‐producing cells is one of the essential processes for higher rAAV vector production. To acquire such cells, a method to identify cells with high expression of transgene protein was developed; however, the relationship between high expression of transgene protein and increased production of rAAV has not been sufficiently studied to date. We used the fluorescent protein ZsGreen1 as a transgene, subdivided the adherent cells according to the intensity of transgene expression protein, and reconfirmed the relationship between rAAV vector production and the amount of intracellular plasmid. We found that the amount of rAAV produced was not correlated with the intensity of expression of the transgene protein but may be correlated with the amount of intracellular plasmid. We also found that cells with high expression of transgene protein might not necessarily produce large amounts of rAAV vector. Based on these results, it was suggested that intracellular plasmid copy number could be used as a new marker for cells producing high levels of rAAV vector.Wiley, 2023年06月, Engineering Reports, 6(1) (1)[査読有り]研究論文（学術雑誌）
- Recombinant adeno-associated virus (rAAV) is a viral vector commonly used in gene therapy. Residual host cell DNA is an impurity that has been associated with the risk of infection and oncogenicity. Thus, it needs to be monitored for quality control. We aimed to develop a droplet digital polymerase chain reaction (ddPCR) method targeting 18S ribosomal RNA (rRNA) genes to quantitate residual host cell DNA. The copy number of the 18S rRNA gene was determined using two sets of primer pairs for 116- and 247-bp amplicons sharing the C-terminus. For conversion of the copy number of the 18S rRNA gene into the mass concentration of genomic DNA, the accurate copy number of 18S rRNA genes in HEK293 genomic DNA was determined by comparison with copy numbers of three reference genes (EIF5B, DCK, and HBB). Results showed that 88.6-97.9% of HEK293 genomic DNA spiked into rAAV preparations was recovered. The ddPCR-based assay was applied to rAAV preparations to quantitate residual host cell DNA as an impurity. Our findings indicate that the assay can be used for the quantitation and size distribution of residual host cell DNA in rAAV products.2023年06月, Human gene therapy, 34(11-12) (11-12), 578 - 585, 英語, 国際誌[査読有り]研究論文（学術雑誌）
- Virus-like particles (VLPs) are hollow nanoparticles composed of recombinant viral surface proteins without a virus genome. In the present study, we investigated the production of influenza VLPs using recombinant insect cells. DNA fragments encoding influenza A virus hemagglutinin (HA) and matrix protein 1 (M1) were cloned with the Drosophila BiP signal sequence in plasmid vectors containing a blasticidin and a neomycin resistance gene, respectively. After Trichoplusia ni BTI-TN-5B1-4 (High Five) cells were co-transfected with a pair of constructed plasmid vectors, stably transformed cells were established via incubation with blasticidin and G418. Western blot analyses showed that recombinant High Five cells secreted HA and M1 proteins into the culture supernatant. Immunoprecipitation of the culture supernatant with an anti-HA antibody and transmission electron microscopy suggested that secreted HA and M1 proteins were in a particulate structure with a morphology similar to that of an influenza virus. Hemagglutination assay indicated that expressed HA molecules retained hemagglutination activity. In a shake-flask culture, recombinant cells achieved a high HA yield (≈ 10 μg/ml) comparable to the yields obtained using the baculovirus-insect cell system. Recombinant insect cells may serve as excellent platforms for the efficient production of influenza VLPs for use as safe and effective vaccines and diagnostic antigens.Elsevier BV, 2020年11月, Biochemical Engineering Journal, 163, 107757 - 107757, 英語, 国際誌[査読有り]研究論文（学術雑誌）
- Insect cells have recently proven to be an excellent platform for the high-level production of functional recombinant proteins. Autophagy is an important mechanism that promotes cell survival by eliminating damaged organelles and protein aggregates, and it also may influence recombinant protein production. In the present study, we compared the effects that autophagy inducers rapamycin, everolimus, and lithium chloride exert on recombinant lepidopteran insect cells that secrete an engineered antibody molecule. Compared with nontreatment, treatment with either rapamycin or everolimus prolonged cell growth to allow high cell density, improved viability in the declining phase, and then increased the yield of secreted antibodies. These positive effects appeared to be induced via autophagy since autophagosomes were clearly detected, particularly in cells treated with rapamycin or everolimus. Unlike rapamycin, another autophagy inducer, FK506, was ineffective in insect cells. The addition of an appropriate autophagy inducer may be effective in increasing the productivity of recombinant proteins in insect cells.2020年09月, Cytotechnology, 73(3) (3), 299 - 305, 英語, 国際誌[査読有り]研究論文（学術雑誌）
- Antibody Fab fragments consist of heavy chain (Hc) and light chain (Lc) polypeptides assembled with a disulphide bond. The production of a recombinant Fab fragment requires the simultaneous expression of two genes encoding both an Hc and an Lc in the same host cell. In the present study, we investigated the production of Fab fragments in lepidopteran insect cells using a bicistronic plasmid vector carrying the Hc and Lc genes linked with a 2A self-cleaving peptide sequence from the porcine teschovirus-1. We also examined the arrangement of a GSG spacer sequence and a furin cleavage site sequence with the 2A sequence. Western blot analysis and enzyme-linked immunosorbent assay (ELISA) of culture supernatants showed that Trichoplusia ni BTI-TN-5B1-4 (High Five) cells transfected with a plasmid in which the Hc and Lc genes were joined by the 2A sequence successfully secreted Fab fragments with antigen-binding activity after self-cleavage of the 2A peptide. The GSG linker enhanced 2A cleavage efficiency, and the furin recognition site was useful for removal of 2A residues from the Hc. Transfection with a single plasmid that contained sequences for GSG, the furin cleavage site, GSG, and the 2A peptide between the Hc and Lc genes exhibited a higher productivity than co-transfection with a set of plasmids separately carrying the Hc or Lc gene. These results demonstrate that bicistronic expression with the appropriate combination of a furin recognition site, GSG linkers, and a 2A peptide may be an effective way to efficiently produce recombinant antibody molecules in insect cells.2020年08月, Journal of bioscience and bioengineering, 130(2) (2), 205 - 211, 英語, 国内誌[査読有り]研究論文（学術雑誌）
- Springer Science and Business Media LLC, 2008年08月, Nature Medicine, 14(9) (9), 939 - 948[査読有り]研究論文（学術雑誌）
- Purpose Docetaxel is one of the most effective anticancer drugs available in the treatment of breast cancer. Nearly half of the treated patients, however, do not respond to chemotherapy and suffer from side effects. The ability to reliably predict a patient's response based on tumor gene expression will improve therapeutic decision making and save patients from unnecessary side effects. Patients and Methods A total of 44 breast tumor tissues were sampled by biopsy before treatment with docetaxel, and the response to therapy was clinically evaluated by the degree of reduction in tumor size. Gene expression profiling of the biopsy samples was performed with 2,453 genes using a high-throughput reverse transcriptase polymerase chain reaction technique. Using genes differentially expressed between responders and nonresponders, a diagnostic system based on the weighted-voting algorithm was constructed. Results This system predicted the clinical response of 26 previously unanalyzed samples with over 80% accuracy, a level promising for clinical applications. Diagnostic profiles in nonresponders were characterized by elevated expression of genes controlling the cellular redox environment (ie, redox genes, such as thioredoxin, glutathione-S-transferase, and peroxiredoxin). Overexpression of these genes protected cultured mammary tumor cells from docetaxel-induced cell death, suggesting that enhancement of the redox system plays a major role in docetaxel resistance. Conclusion These results suggest that the clinical response to docetaxel can be predicted by gene expression patterns in biopsy samples. The results also suggest that one of the molecular mechanisms of the resistance is activation of a group of redox genes.American Society of Clinical Oncology (ASCO), 2005年01月, Journal of Clinical Oncology, 23(3) (3), 422 - 431[査読有り]研究論文（学術雑誌）
- Molecular classification of primary breast tumors possessing distinct prognostic properties.The natural progression of breast cancer differs greatly between patients; the precise prediction of this disease course will improve the efficacy of therapeutics. Gene expression profiling may elucidate the undiscovered biological variations between seemingly similar cancers, leading to a new cancer classification system valuable in accurate diagnosis. The expression levels of 2412 genes, derived from 98 cancer samples, were precisely recorded by a high throughput RT-PCR technique, adapter-tagged competitive PCR. Subsequent cluster analysis revealed a molecular profile, correlating with estrogen receptor levels and the presence of lymph node metastases. We analyzed 301 cancer samples for the expression patterns of 21 genes critical in this categorization. The classification of the samples into three major groups was verified utilizing principal component analysis. This molecular classification system correlated significantly with early recurrence, independent of lymph node status. This malignant potential is associated with the expression levels of a group of genes, which comprise a set of candidates potentially useful in diagnostic prediction. These genes and the associated control mechanisms may also be effective therapeutic targets.2002年01月, Human molecular genetics, 11(2) (2), 199 - 206, 英語, 国際誌[査読有り]研究論文（学術雑誌）
- 2009年04月, NEURO-ONCOLOGY, 11(2) (2), 219 - 219, 英語USING GENE EXPRESSION PROFILING TO IDENTIFY A PROGNOSTIC MOLECULAR SPECTRUM IN GLIOMAS研究発表ペーパー・要旨（国際会議）
- 2008年10月, NATURE MEDICINE, 14(10) (10), 1128 - 1128, 英語RPN2 gene confers docetaxel resistance in breast cancer (vol 14, pg 939, 2008)その他
- 第70回日本生物工学会大会, 2018年09月, 日本語, 日本生物工学会大会, 吹田, 国内会議昆虫細胞における2Aペプチドを用いた抗体生産口頭発表（一般）
- 化学工学会第50回秋季大会, 2018年09月, 英語, 化学工学会, 鹿児島, 国内会議ゲノム編集技術を用いた組換え昆虫細胞の作製ポスター発表
- 化学工学会第83年会, 2018年03月, 日本語, 化学工学会, 吹田, 国内会議昆虫細胞によるインフルエンザウイルス抗原タンパク質の生産ポスター発表
- 化学工学会第49回秋季大会, 2017年09月, 日本語, 化学工学会, 名古屋, 国内会議昆虫細胞による2Aペプチドを用いた抗体タンパク質の生産ポスター発表
- 日本学術振興会, 科学研究費助成事業, 奨励研究, 兵庫医科大学, 2016年04月01日 - 2017年03月31日胆管細胞癌におけるＢ型肝炎ウイルスゲノム組み込み解析研究
- 日本学術振興会, 科学研究費助成事業, 基盤研究(C), 武庫川女子大学, 2012年04月01日 - 2015年03月31日筋疾患予防型トレーニングに遺伝子解析を活用しようスポーツ選手にとって靱帯損傷などの故障は人生設計の変更を伴う重大事件であるが、現在までにそれら受傷しやすさを予測するような遺伝子は発見されていない。今回、靱帯損傷などの故障リスクを予測し予防することを目的として、本学運動部99名を対象にスポーツや筋肉に関わる遺伝子型検査およびアンケート調査を実施した。その結果ACTN3遺伝子の577Rは577Xよりも有意に筋疾患歴を持つことが判明した。この結果より、ACTN3遺伝子多型を調べることで自身の受傷リスクが分かる可能性が示唆された。今後この結果をもとに、筋疾患を回避するためのトレーニング方法を検討していく予定である。
- 日本学術振興会, 科学研究費助成事業, 基盤研究(C), 武庫川女子大学, 2011年 - 2013年次世代への遺伝子および遺伝子検査教育のための研究本研究における教育活動として、当初の高等学校での出前講義・実験および大学での初年次教育に加え、小学生とその保護者、飲料関連企業、薬局などで、遺伝子検査を含むゲノム科学リテラシーセミナーを実施し、遺伝子に関する教育方法および解析技術を改良した。アルコール遺伝子検査を行った大学では３年次以降での追跡調査も開始した。教育方法の改善に加えて、高校での遺伝子実験法の改良により、生徒自身が実験を主導できる実験系プロトコールを開発し、個人情報取り扱いを含めた高校生主体プロジェクトの開始に向けた検討を行った。また大学でのアドバンス科目用として遺伝子情報取り扱いのための基礎技術も開発した。
- 日本学術振興会, 科学研究費助成事業, 若手研究(B), 地方独立行政法人大阪府立病院機構大阪府立成人病センター(研究所), 2005年 - 2006年染色体の増幅・欠失の網羅的検出解析による多発及び再発肝細胞癌の遺伝子診断の確立肝細胞癌では、外科的な根治的切除後や局所治療後も、5年以内に約80%で再発が見られる。これは微少肝内転移(IM)や異時性多中心性発癌(MC)が原因と考えられるが、IMとMCでは治療方針が異なるために、この両者を見分けることは癌のオーダーメイドな治療を行う上で臨床上重要であると考えられる。本研究では、染色体におけるゲノムコピー数を測定する手法を開発し、その手法を用いてIMとMCの染色体異常を同定し、その違いを解明することを目的とする。 まず、今年度はPCRベースでの染色体ゲノムコピー数の変化を同定する手法(Competitive Genome PCR法;CGP法と命名)の開発を行った。CGP法はPCRで行うため、簡便で臨床的にも使いやすいと考えられる。現在までに本手法を開発し確立したが、本手法の感度を検定するため、以下の解析を行った。まず、X染色体のコピー数の異なるcell line(46XY(1X)、46XX(2X)、47XXX(3X)、48XXXX(4X)、49XXXXX(5X)の5種類の核型をもつcell line)からのgenomic DNAを用いて、46XXをreferenceとし、本手法によりX染色体のコピー数の比較を行ったところ、微小なゲノムコピー数変化も検出でき、高感度で解析が行えることが判明した。また12種類のneuroblastoma cell lineからのgenomic DNAを用いた、MYCN遺伝子近傍の増幅領域の同定をするなど、実用的な解析が可能であることも明らかとした。この手法は、昨年11月に国内で特許申請を行ったほか、現在、論文の投稿も完了している。現在は肝細胞癌を用いて、IM,MCの遺伝子診断を行うべく、プライマーを設計し、順次解析を行っているところである。