細胞修飾による肝特異的経静脈的間葉系幹細胞移植法の開発

2019 Fiscal Year Annual Research Report

• Project/Area Number: 17H02100
• Research Institution: Osaka University
• Principal Investigator: 山下 静也 大阪大学, 産業科学研究所, 招へい教授 (60243242)
• Co-Investigator(Kenkyū-buntansha): 小関 正博 大阪大学, 医学系研究科, 助教 (10467582) ; 黒田 俊一 大阪大学, 産業科学研究所, 教授 (60263406)
• Project Period (FY): 2017-04-01 – 2020-03-31
• Keywords: 幹細胞移植 / 糖鎖修飾 / 家族性高コレステロール血症 / 細胞治療 / 肝特異的

Outline of Annual Research Achievements

前年度までの基礎検討で、既製品の化合物同士をアビジンビオチン結合で結合させて細胞修飾を行ったが、細胞が凝集する傾向が強かった。そこで脂肪酸に糖鎖Xを直接結合させた糖鎖脂質誘導体Yを作成して細胞修飾し、蛍光色素でラベルした。条件等を行ったが、糖鎖脂質誘導体Yのみでの修飾では細胞生存率は問題なかったが、蛍光色素でラベルすると単独でも細胞生存率が不良で、糖鎖脂質誘導体Yでの修飾と蛍光色素のラベルを併用するとさらに細胞障害性が強く、ヌードマウスの尾静脈から注入してin vivo imagingで観察する系で観察することは困難であることが分かった。そこで、マウスをC57Bl/6に変更し、糖鎖脂質誘導体Yでヒト細胞株由来の細胞を修飾したものを作成し尾静脈より移植し、サクリファイス後、肝切片を作成し抗ヒトTCR抗体で染色して、生着を評価する実験を継続した。我々は、再生医療等安全性確保法にもとづいてヒト脂肪組織由来間葉系幹細胞製剤を作成し、ヒトに移植した経験があり、そのプロトコールを応用して、マウスの脂肪組織から間葉系幹細胞分画を抽出したが、マウスの脂肪組織中の間葉系幹細胞はヒトに比べて著しく少なく培養を繰り返しても移植用の十分な細胞数を得ることが困難であった。
一方で将来的な、細胞治療の被験者候補である、家族性高コレステロール血症の患者は、脂質専門外来に多くの紹介があり、100人以上に対して脂質異常症遺伝子診断パネルを作成して遺伝子解析を実施し、重複遺伝子変異が存在するなどの知見を得ることができた。

Research Progress Status

令和元年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

令和元年度が最終年度であるため、記入しない。

Research Products

• [Journal Article] Pemafibrate, a New Selective PPARα Modulator: Drug Concept and Its Clinical Applications for Dyslipidemia and Metabolic Diseases. (2020)
  • Author(s): Yamashita S, Masuda D, Matsuzawa Y
  • Journal Title: Curr Atheroscler Rep Volume: 22 Pages: 5
  • DOI: 10.1007/s11883-020-0823-5
  • Peer Reviewed / Open Access
• [Journal Article] Omega-3 fatty acid ethyl esters improve low-density lipoprotein subclasses without increasing low-density lipoprotein-cholesterol levels: A phase 4, randomized study. (2020)
  • Author(s): Masuda D, Miyata Y, Matsui S, Yamashita S
  • Journal Title: .Atherosclerosis. Volume: 292 Pages: 163-170
  • DOI: 10.1016/j.atherosclerosis.2019.11.014.
  • Peer Reviewed / Open Access
• [Journal Article] Progranulin deficiency leads to enhanced age-related cardiac hypertrophy through complement C1q-induced β-catenin activation. (2020)
  • Author(s): Zhu Y, Ohama T, Kawase R, Chang J, Inui H, Kanno K, Okada T, Masuda D, Koseki M, Nishida M, Sakata Y, Yamashita S
  • Journal Title: J Mol Cell Cardiol. Volume: 138 Pages: 197-211
  • DOI: 10.1016/j.yjmcc.2019.12.009
  • Peer Reviewed / Open Access
• [Journal Article] Residual vascular risk in diabetes - Will the SPPARM alpha concept hold the key? (2019)
  • Author(s): Fruchart JC, Santos RD, Yamashita S, Libby P; International Atherosclerosis Society/R3i Foundation Consensus Panel.Fruchart JC, et al.
  • Journal Title: Diabetes Metab Syndr Volume: 13 Pages: 2723-2725
  • DOI: 10.1016/j.dsx.2019.05.034
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Yamashita S, Masuda D, Matsuzawa Y.Yamashita S, et al (2019)
  • Author(s): Yamashita S, Masuda D, Matsuzawa Y.Yamashita S, et al
  • Journal Title: J Atheroscler Thromb Volume: 26 Pages: 389-402
  • DOI: 10.5551/jat.48918
  • Peer Reviewed / Open Access
• [Journal Article] Long-Term Treatment With Evolocumab Among Japanese Patients - Final Report of the OSLER Open-Label Extension Studies. (2019)
  • Author(s): Hirayama A, Yamashita S, Ruzza A, Inomata H, Cyrille M, Lu C, Hamer AW, Yoshida M, Kiyosue A, Teramoto T.Hirayama A, et al.
  • Journal Title: Circ J. Volume: 83 Pages: 971-977
  • DOI: 10.1253/circj.CJ-19-0139
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Long-Term Efficacy and Safety of Pemafibrate, a Novel Selective Peroxisome Proliferator-Activated Receptor-α Modulator (SPPARMα), in Dyslipidemic Patients with Renal Impairment. (2019)
  • Author(s): Yokote K, Yamashita S, Arai H, Araki E, Suganami H, Ishibashi S, Of The K-Study Group OB.Yokote K, et al.
  • Journal Title: Int J Mol Sci. Volume: 20 Pages: 706
  • DOI: 10.3390/ijms20030706
  • Peer Reviewed / Open Access
• [Journal Article] Efficacy and safety of pemafibrate in people with type 2 diabetes and elevated triglyceride levels: 52-week data from the PROVIDE study. (2019)
  • Author(s): Araki E, Yamashita S, Arai H, Yokote K, Satoh J, Inoguchi T, Nakamura J, Maegawa H, Yoshioka N, Tanizawa Y, Watada H, Suganami H, Ishibashi S.Araki E, et al
  • Journal Title: Diabetes Obes Metab. Volume: 21 Pages: 1737-1744
  • DOI: 10.1111/dom.13686
  • Peer Reviewed / Open Access
• [Journal Article] Efficacy and Safety of Pemafibrate, a Novel Selective Peroxisome Proliferator-Activated Receptor α Modulator (SPPARMα): Pooled Analysis of Phase 2 and 3 Studies in Dyslipidemic Patients with or without Statin Combination. (2019)
  • Author(s): Yamashita S, Arai H, Yokote K, Araki E, Matsushita M, Nojima T, Suganami H, Ishibashi S.Yamashita S, et al
  • Journal Title: Int J Mol Sci Volume: 20 Pages: 5537
  • DOI: 10.3390/ijms20225537
  • Peer Reviewed / Open Access
• [Journal Article] Rivaroxaban Suppresses the Progression of Ischemic Cardiomyopathy in a Murine Model of Diet-Induced Myocardial Infarction. (2019)
  • Author(s): Liu J, Nishida M, Inui H, Chang J, Zhu Y, Kanno K, Matsuda H, Sairyo M, Okada T, Nakaoka H, Ohama T, Masuda D, Koseki M, Yamashita S, Sakata Y.
  • Journal Title: J Atheroscler Thromb Volume: 26 Pages: 915-930
  • DOI: 10.5551/jat.48405
  • Peer Reviewed / Open Access
• [Journal Article] Comparative aspects of the care of familial hypercholesterolemia in the "Ten Countries Study". (2019)
  • Author(s): Pang J, Chan DC, Hu M, Muir LA, Kwok S, Charng MJ, Florkowski CM, George PM, Lin J, Loi DD, Marais AD, Nawawi HM, Gonzalez-Santos LE, Su TC, Truong TH, Santos RD, Soran H, Tomlinson B, Yamashita S, Ademi Z, Watts GF.
  • Journal Title: J Clin Lipidol. Volume: 13 Pages: 287-300
  • DOI: 10.1016/j.jacl.2019.01.009
  • Peer Reviewed / Open Access
• [Journal Article] Real-World Data to Identify Hypercholesterolemia Patients on Suboptimal Statin Therapy. (2019)
  • Author(s): Kajinami K, Ozaki A, Tajima Y, Yamashita S, Arai H, Teramoto T
  • Journal Title: J Atheroscler Thromb. Volume: 26 Pages: 408-431
  • DOI: 10.5551/jat.46201
  • Peer Reviewed / Open Access
• [Journal Article] Cultural Barriers in the Treatment of Dyslipidemia: A Survey of Japanese Physician Attitudes. (2019)
  • Author(s): Yamashita S, Masuda D, Arai H, Matsuzawa Y
  • Journal Title: J Atheroscler Thromb. Volume: 26 Pages: 154-169
  • DOI: 10.5551/jat.44677
  • Peer Reviewed / Open Access
• [Journal Article] Shotgun proteomic analysis reveals proteome alterations in HDL of patients with cholesteryl ester transfer protein deficiency. (2019)
  • Author(s): Okada T, Ohama T, Takafuji K, Kanno K, Matsuda H, Sairyo M, Zhu Y, Saga A, Kobayashi T, Masuda D, Koseki M, Nishida M, Sakata Y, Yamashita S.
  • Journal Title: J Clin Lipidol Volume: 13 Pages: 317-325
  • DOI: 10.1016/j.jacl.2019.01.002.
  • Peer Reviewed / Open Access
• [Presentation] A selective PPARα modulator, pemafibrate reduced hepatic cholesterol content and inflammatory cell infiltration in atherogenic-diet induced steatohepatitis mouse model (2020)
  • Author(s): Kanno K, Koseki M, Saga A, Chang J, Okada T, Inui H, Asaji M, Zhu Y, Ohama T, Nishida M, Yamashita S, Sakata Y.
  • Organizer: Keystone Symposia 2020
  • Int'l Joint Research
• [Presentation] The Current Status of the Treatment of FH Patients and the Achievement of LDL-C Target Value in the Lipid Clinic in Osaka University Hospital (2019)
  • Author(s): Koseki M, Okada T, Kanno K, Ohama T, Nishida M, Yamashita S, Sakata Y.
  • Organizer: FH Global Summit
  • Int'l Joint Research
• [Presentation] Pemafibrate regulate intestinal lipid absorption in diet-induced nonalcoholic steatohepatitis model in mice (2019)
  • Author(s): 冠野 昂太郎、小関 正博、嵯峨 礼美、常 久洋、岡田 健志、乾 洋勉、朝治 真澄、朱 穎竑、大濱 透、西田 誠、山下 静也、坂田 泰史
  • Organizer: AHA2019
  • Int'l Joint Research
• [Presentation] The Predominance of Th1 and Th17 During the Process of Myocardial Infarction in Non-surgical MI model, SR-BI KO/ApoeR61h/h mice (2019)
  • Author(s): Inui H, Nishida M, Asaji M, Saga A, Kanno K , Chang J, Zhu Y , Okada T, Ohama T , Koseki M, Yamashita S, Sakata Y.
  • Organizer: AHA2019
  • Int'l Joint Research
• [Presentation] Pgrn Suppresses Pressure Overload-induced Cardiac Hypertrophy And Heart Failure In Mice Via Inhibiting The Activation Of β-catenin (2019)
  • Author(s): Zhu Y, Ohama T, Inui H, Kanno K, Chang J, Okada T, Koseki M, Nishida M, Sakata Y, Yamashita S.
  • Organizer: AHA2019
  • Int'l Joint Research
• [Remarks] 大阪大学 大学院医学系研究科 循環器内科学循環器脂質・動脈硬化グループ
  • URL: http://www.cardiology.med.osaka-u.ac.jp/?page_id=33160
• [Remarks] 家族性高コレステロール血症 大阪大学 大学院医学系研究科 循環器内科学 循環器脂質・動脈硬化研究室
  • URL: http://fh-osaka-univ.org/