Creation of Knock-out rabbit models for the study of atherosclerosis

• Project/Area Number: 15H04718
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Experimental pathology
• Research Institution: University of Yamanashi
• Principal Investigator: FAN Jianglin 山梨大学, 大学院総合研究部, 教授 (60272192)
• Co-Investigator(Kenkyū-buntansha): 北嶋 修司 佐賀大学, 総合分析実験センター, 准教授 (70284643)
• Project Period (FY): 2015-04-01 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥14,430,000 (Direct Cost: ¥11,100,000、Indirect Cost: ¥3,330,000); Fiscal Year 2018: ¥3,640,000 (Direct Cost: ¥2,800,000、Indirect Cost: ¥840,000); Fiscal Year 2017: ¥3,640,000 (Direct Cost: ¥2,800,000、Indirect Cost: ¥840,000); Fiscal Year 2016: ¥3,510,000 (Direct Cost: ¥2,700,000、Indirect Cost: ¥810,000); Fiscal Year 2015: ¥3,640,000 (Direct Cost: ¥2,800,000、Indirect Cost: ¥840,000)
• Keywords: リポ蛋白代謝 / 高脂血症 / 動脈硬化 / 動物モデル / リポ蛋白 / 脂質代謝 / 遺伝子改変ウサギ / ノックアウト / ウサギモデル / 脂質 / 病理学 / 循環器 / 応用動物

Outline of Final Research Achievements

Apolipoprotein CIII (apoCIII) is a constituent of all lipoproteins except LDLs and mediates the metabolism of triglyceride (TG)-rich lipoproteins through inhibition of LPL activity. Elevated plasma levels of apoCIII are directly correlated with the plasma TG and increase the cardiovascular risk. However, the pathophysiological functions of apoCIII in vivo have not been fully elucidated. To examine the functional roles of apoCIII in lipoprotein metabolism and atherosclerosis, we generated both trangesgenic (Tg) and apoCIII knockout (KO) rabbits. Tg rabbits showed high levels of plasma TG whereas apoCIII KO rabbits exhibited lower plasma levels of TG than those of wild-type (WT) rabbits while total cholesterol and HDL-cholesterol levels were unchanged. Analysis of lipoproteins revealed that reduced plasma TG levels in KO rabbits were accompanied by significant reduction of VLDLs and intermediate-density lipoproteins (IDLs).

Academic Significance and Societal Importance of the Research Achievements

動脈硬化とその合併症である心筋梗塞、脳卒中といった疾患は、日本を始めとした先進国における死亡原因の約30%を占め、人口の高齢化が加速する中で、今や医学領域のみならず社会経済的にも最も重要な課題となっている。新規アポCIII遺伝子改変ウサギモデルの確立は、動脈硬化の研究に限らず、脂質代謝、高脂血症の研究及び新薬の開発に新たな研究ツールとしての利用が期待される。

Research Products

• [Int'l Joint Research] ミシガン大学/心臓血管研究所(米国)
• [Int'l Joint Research] 西安交通大学/動脈硬化研究所(中国)
• [Int'l Joint Research] 中国科学院/都市環境健康研究所(中国)
• [Int'l Joint Research] ミシガン大学/心臓血管研究所(米国)
• [Int'l Joint Research] 西安交通大学/動脈硬化研究所(中国)
• [Int'l Joint Research] ミシガン大学/心臓血管研究所(米国)
• [Int'l Joint Research] 西安交通大学/動脈硬化研究所(中国)
• [Journal Article] Generation of hyperlipidemic rabbit models using multiple sgRNAs targeted CRISPR/Cas9 gene editing system (2019)
  • Author(s): Yuan Tingting、Zhong Yi、Wang Yingge、Zhang Ting、Lu Rui、Zhou Minya、Lu Yaoyao、Yan Kunning、Chen Yajie、Hu Zhehui、Liang Jingyan、Fan Jianglin、Cheng Yong
  • Journal Title: Lipids in Health and Disease Volume: 18 Issue: 1 Pages: 6-80
  • DOI: 10.1186/s12944-019-1013-8
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Renovascular Hypertension Aggravates Atherosclerosis in Cholesterol-Fed Rabbits (2019)
  • Author(s): Chen Yajie、Waqar Ahmed?Bilal、Yan Haizhao、Wang Yanli、Liang Jingyan、Fan Jianglin
  • Journal Title: Journal of Vascular Research Volume: 56 Issue: 1 Pages: 28-38
  • DOI: 10.1159/000498897
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Hypoxic and Cold Adaptation Insights from the Himalayan Marmot Genome (2019)
  • Author(s): Bai Liang、Liu Baoning、Ji Changmian、Zhao Sihai、Liu Siyu、Wang Rong、Wang Weirong、Yao Pu、Li Xuming、Fu Xiaojun、Yu Haiyan、Liu Min、Han Fengming、Guan Ning、Liu Hui、Liu Dongyuan、Tao Yuanqing、Wang Zhongdong、Yan Shunsheng、Florant Greg、Butcher Michael T.、Zhang Jifeng、Zheng Hongkun、Fan Jianglin、Liu Enqi
  • Journal Title: iScience Volume: 11 Pages: 505-507
  • DOI: 10.1016/j.isci.2019.01.019
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Treatment of atherosclerosis by traditional Chinese medicine: Questions and quandaries (2018)
  • Author(s): Wang Chuan、Niimi Manabu、Watanabe Teruo、Wang Yanli、Liang Jingyan、Fan Jianglin
  • Journal Title: Atherosclerosis Volume: 277 Pages: 136-144
  • DOI: 10.1016/j.atherosclerosis.2018.08.039
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Hypertension Enhances Advanced Atherosclerosis and Induces Cardiac Death in Watanabe Heritable Hyperlipidemic Rabbits (2018)
  • Author(s): Ning Bo、Chen Yajie、Waqar Ahmed Bilal、Yan Haizhao、Shiomi Masashi、Zhang Jifeng、Chen Y. Eugene、Wang Yanli、Itabe Hiroyuki、Liang Jingyan、Fan Jianglin
  • Journal Title: The American Journal of Pathology Volume: 188 Issue: 12 Pages: 2936-2947
  • DOI: 10.1016/j.ajpath.2018.08.007
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Spontaneous severe hypercholesterolemia and atherosclerosis lesions in rabbits with deficiency of low-density lipoprotein receptor (LDLR) on exon 7 (2018)
  • Author(s): Lu Rui、Yuan Tingting、Wang Yingge、Zhang Ting、Yuan Yuguo、Wu Daijin、Zhou Minya、He Zhengyi、Lu Yaoyao、Chen Yajie、Fan Jianglin、Liang Jingyan、Cheng Yong
  • Journal Title: EBioMedicine Volume: 36 Pages: 29-38
  • DOI: 10.1016/j.ebiom.2018.09.020
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Sphingolipid de novo biosynthesis is essential for intestine cell survival and barrier function (2018)
  • Author(s): Li Zhiqiang、Kabir Inamul、Tietelman Gladys、Huan Chongmin、Fan Jianglin、Worgall Tilla、Jiang Xian-Cheng
  • Journal Title: Cell Death & Disease Volume: 9 Issue: 2 Pages: 173-183
  • DOI: 10.1038/s41419-017-0214-1
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Detection of potential new biomarkers of atherosclerosis by probe electrospray ionization mass spectrometry (2018)
  • Author(s): Johno Hisashi、Yoshimura Kentaro、Mori Yuki、Kimura Tokuhide、Niimi Manabu、Yamada Masaki、Tanigawa Tetsuo、Fan Jianglin、Takeda Sen
  • Journal Title: Metabolomics Volume: 14 Issue: 4 Pages: 1-6
  • DOI: 10.1007/s11306-018-1334-z
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Principles and Applications of Rabbit Models for Atherosclerosis Research (2018)
  • Author(s): Fan J, Chen Y, Yan H, Niimi M, Wang Y, Liang J.
  • Journal Title: Journal of Atherosclerosis and Thrombosis Volume: 25 Issue: 3 Pages: 213-220
  • DOI: 10.5551/jat.RV17018
  • NAID: 130006407212
  • ISSN: 1340-3478, 1880-3873
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Genomic and Transcriptomic Analysis of Hypercholesterolemic Rabbits: Progress and Perspectives (2018)
  • Author(s): Fan Jianglin、Chen Yajie、Yan Haizhao、Liu Baoning、Wang Yanli、Zhang Jifeng、Chen Y.、Liu Enqi、Liang Jingyan
  • Journal Title: International Journal of Molecular Sciences Volume: 19 Issue: 11 Pages: 3512-3512
  • DOI: 10.3390/ijms19113512
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Glutathione inhibits antibody and complement-mediated immunologic cell injury via multiple mechanisms. (2017)
  • Author(s): Zhang Z, Zhang X, Fang X, Niimi M, Huang Y, Piao H, Gao S, Fan J, Yao J.
  • Journal Title: Redox Biol. Volume: 12 Pages: 571-581
  • DOI: 10.1016/j.redox.2017.03.030
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Carbenoxolone inhibits TRPV4 channel-initiated oxidative urothelial injury and ameliorates cyclophosphamide-induced bladder dysfunction (2017)
  • Author(s): Zhang X,Gao S,Tanaka M,Zhanq Z,Huanq Y,Mitsui T,Kamiyama M,Koizumi S,Fan J,Takeda M,Yao J
  • Journal Title: J Cell Mol Med Volume: 2017 Issue: 9 Pages: 346-346
  • DOI: 10.1111/jcmm.13100
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Deficiency of Cholesteryl Ester Transfer Protein Protects Against Atherosclerosis in Rabbits. (2017)
  • Author(s): Zhang, J. Niimi, M. Yang, D. Liang, J. Xu, J. Kimura, T. Mattew, A. V. Guo, Y. Fan, Y. Zhu, T. Song, J. Ackermann, R. Koike, Y. Schwendeman, A. Lai, L. Pennathur, S. Garcia-Barrio, M. Fan, J. Chen, Y. E.
  • Journal Title: Arterioscler Thromb Vasc Biol Volume: in press Issue: 6 Pages: 1068-1075
  • DOI: 10.1161/atvbaha.117.309114
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Comparative studies of three cholesteryl ester transfer proteins and their interactions with known inhibitors. (2017)
  • Author(s): Wang Z, Niimi M, Ding Q, Liu Z, Wang L, Zhang J, Xu J, Fan J.
  • Journal Title: PLoS One. Volume: 12 Issue: 8 Pages: e0180772-e0180772
  • DOI: 10.1371/journal.pone.0180772
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Increased hepatic expression of endothelial lipase inhibits cholesterol diet induced hypercholesterolemia and atherosclerosis in transgenic rabbits (2017)
  • Author(s): Wang, C., Nishijima, K., Kitajima, S., Niimi, M., Yan, H., Chen, Y., Ning, B., Matsuhisa, F., Liu, E., Zhang, J., Chen, Y. E., Fan. J.
  • Journal Title: Arterioscler. Thromb. Vasc. Biol. Volume: 37 Issue: 7 Pages: 1282-1289
  • DOI: 10.1161/atvbaha.117.309139
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Bre Enhances Osteoblastic Differentiation by Promoting the Mdm2-Mediated Degradation of p53 (2017)
  • Author(s): Jin Fujun、Wang Yiliang、Wang Xiaojing、Wu Yanting、Wang Xiaoyan、Liu Qiuying、Zhu Yexuan、Liu Enqi、Fan Jianglin、Wang Yifei
  • Journal Title: STEM CELLS Volume: 35 Issue: 7 Pages: 1760-1772
  • DOI: 10.1002/stem.2620
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Overexpression of Cholesteryl Ester Transfer Protein Increases Macrophage-Derived Foam Cell Accumulation in Atherosclerotic Lesions of Transgenic Rabbits (2017)
  • Author(s): Gao Shoucui、Wang Xiaojing、Cheng Daxing、Li Jiayan、Li Lu、Ran Linwu、Zhao Sihai、Fan Jianglin、Liu Enqi
  • Journal Title: Mediators Inflamm Volume: 2017 Pages: 1-9
  • DOI: 10.1155/2017/3824276
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Mediator 1 Is Atherosclerosis Protective by Regulating Macrophage PolarizationHighlights (2017)
  • Author(s): Bai Liang、Li Zhao、Li Qianwei、Guan Hua、Zhao Sihai、Liu Ruihan、Wang Rong、Zhang Jin、Jia Yuzhi、Fan Jianglin、Wang Nanping、Reddy Janardan K.、Shyy John Y.-J.、Liu Enqi
  • Journal Title: Arteriosclerosis, Thrombosis, and Vascular Biology Volume: 37 Issue: 8 Pages: 1470-1481
  • DOI: 10.1161/atvbaha.117.309672
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] AMPK Suppresses Connexin43 Expression in the Bladder and Ameliorates Voiding Dysfunction in Cyclophosphamide-induced Mouse Cystitis (2016)
  • Author(s): Zhang X, Yao J, Gao K, Chi Y, Mitsui T, Ihara T, Sawada N, Kamiyama M, Fan J, Takeda M
  • Journal Title: Sci Rep. Volume: 6 Issue: 1 Pages: 19708-19708
  • DOI: 10.1038/srep19708
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Protein inhibitor of activated stat3 suppresses oxidized ldl-induced cell responses during atherosclerosis in apolipoprotein e-deficient mice. (2016)
  • Author(s): Wang R, Zhang Y, Xu L, Lin Y, Yang X, Bai L, Chen Y, Zhao S, Fan J, Cheng X, Liu E.
  • Journal Title: Sci Rep Volume: 6 Issue: 1 Pages: 36790-36790
  • DOI: 10.1038/srep36790
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Combined b, t and nk cell deficiency accelerates atherosclerosis in balb/c mice. (2016)
  • Author(s): Cheng F, Twardowski L, Reifenberg K, Winter K, Canisius A, Pross E, Fan J, Schmitt E, Shultz LD, Lackner KJ, Torzewski M.
  • Journal Title: PloS One Volume: 11 Issue: 8 Pages: e0157311-e0157311
  • DOI: 10.1371/journal.pone.0157311
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Dietary cocoa powder improves hyperlipidemia and reduces atherosclerosis in apoe deficient mice through the inhibition of hepatic endoplasmic reticulum stress. (2016)
  • Author(s): Guan H, Lin Y, Bai L, An Y, Shang J, Wang Z, Zhao S, Fan J, Liu E.
  • Journal Title: Mediators of inflammation Volume: 2016 Pages: 1937572-1937572
  • DOI: 10.1155/2016/1937572
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Plasma High-Mannose and Complex/Hybrid N-Glycans Are Associated with Hypercholesterolemia in Humans and Rabbits. (2016)
  • Author(s): Bai L, Li Q, Li L, Lin Y, Zhao S, Wang W, Wang R, Li Y, Yuan J, Wang C, Wang Z, Fan J, Liu E.
  • Journal Title: PloS one Volume: 11 Issue: 3 Pages: e0146982-e0146982
  • DOI: 10.1371/journal.pone.0146982
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Hyperlipidemia-associated variations and expression patterns revealed by whole-genome and transcriptome sequencing of rabbit models" (2016)
  • Author(s): Wang Z, Zhang J, Li H,Li S, Ning B, Wang Z, Koike T, Shiomi M, Fan L, Chen YE, Li Y.
  • Journal Title: Scientific Reports Volume: in press Issue: 1 Pages: 00-00
  • DOI: 10.1038/srep26942
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Production of human apolipoprotein(a) transgenic NIBS miniature pigs by somatic cell nuclear transfer (2016)
  • Author(s): Shimatsu Y, Horii W, Nunoya T, Iwata A, Fans J, Ozawa M.
  • Journal Title: Experimental Animals Volume: 65 Issue: 1 Pages: 37-43
  • DOI: 10.1538/expanim.15-0057
  • NAID: 130005125853
  • ISSN: 0007-5124, 1341-1357, 1881-7122
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] ApoE knockout rabbits: A novel model for the study of human hyperlipidemia. (2016)
  • Author(s): Niimi M, Yang D, Kitajima S, Ning B, Wang C, Li S, Liu E, Zhang J, Eugene Chen Y, Fan J.
  • Journal Title: Atherosclerosis Volume: 245 Pages: 187-193
  • DOI: 10.1016/j.atherosclerosis.2015.12.002
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Angiotensin II Destabilizes Coronary Plaques in Watanabe Heritable Hyperlipidemic Rabbits. (2016)
  • Author(s): Li S, Wang YN, Niimi M, Ning B, Chen Y, Kang D, Wang Z, Yu Q, Waqar AB, Liu E, Zhang J, Shiomi M, Chen YE, Fan J.
  • Journal Title: Arterioscler Thromb Vasc Biol Volume: 36 Issue: 5 Pages: 810-6
  • DOI: 10.1161/atvbaha.115.306871
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] LaGomiCs-Lagomorph Genomics Consortium: An International Collaborative Effort for Sequencing the Genomes of an Entire Mammalian Order (2016)
  • Author(s): Fontanesi L, Di Palma F, Flicek P, Smith AT, Thulin CG, Alves PC. LaGomiCs-Lagomorph Genomics Consortium
  • Journal Title: J Hered Volume: 107 Issue: 4 Pages: 295-308
  • DOI: 10.1093/jhered/esw010
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] 動脈硬化病理-known and unknown (2018)
  • Author(s): 範 江林
  • Organizer: 第18回動脈硬化教育フォーラム
• [Presentation] Macrophage-derived matrix metalloprotease-1 accelerates aortic aneurysm formation in transgenic rabbits. (2017)
  • Author(s): 範 江林
  • Organizer: 第49回日本動脈硬化学会総会・学術集会
• [Presentation] Elucidation of Novel Functional Roles of MMPs in Atherosclerosis using Transgenic Rabbits. (2017)
  • Author(s): 範 江林
  • Organizer: 第2回全国（中国）基質生物学学術会議
  • Int'l Joint Research / Invited
• [Presentation] Improved Triglyceride-Rich Lipoprotein Clearance In Human Apolipoprotein A-II Knockin Rabbits. (2017)
  • Author(s): Jiganglin Fan
  • Organizer: ATVB/PVD Scientific Sessions 2017
  • Int'l Joint Research
• [Presentation] The whole-genome and transcriptome sequencing analysis of NZW and JW rabbits and WHHL rabbits. (2016)
  • Author(s): Wang Z, Zhang J, Li H, Shiomi M, Liu E, Chen YE, Li Y, Fan J.
  • Organizer: 第3回日中合同ウサギバイオサイエンスフォーラム
  • Place of Presentation: 佐賀大学鍋島キャンパス（佐賀県佐賀市）
  • Year and Date: 2016-08-06
• [Presentation] Angiotensin II destabilizes coronary plaques in WHHL rabbits. (2016)
  • Author(s): Li S, Wang Y, Miimi M, Ning B, Chen Y, Kang D, Wang Z, Yu Q, Waqar AB, Liu E, Zhang J, Shiomi M, Chen YE, Fan J.
  • Organizer: 第48回日本動脈硬化学会総会・学術集会
  • Place of Presentation: 京王プラザホテル（東京都新宿区）
  • Year and Date: 2016-07-14
• [Presentation] Hepatic overexpression of endothelial lipase reduces plasma HDL levels in transgenic rabbits. (2016)
  • Author(s): Wang C, Nishijima K, Kitajima S, Niimi M, Ning B, Fan J.
  • Organizer: 第48回日本動脈硬化学会総会・学術集会
  • Place of Presentation: 京王プラザホテル（東京都新宿区）
  • Year and Date: 2016-07-14
• [Presentation] Angiotensin II induces cornoary plaque rupture and myocardial infarction in hyperlipidemic rabbits (2015)
  • Author(s): Fan, L.
  • Organizer: The 4th Hsien Wu and Ray Wu symposium
  • Place of Presentation: （Peking, China）North Garden Hotel
  • Year and Date: 2015-08-10
  • Int'l Joint Research / Invited
• [Presentation] ApoE knockout rabbits: a novel model for the study of human hyperlipidemia (2015)
  • Author(s): Niimi, M, Ning B, Wang C, Li S, Zhang J, Chen, YE, Fan L.
  • Organizer: 第47回日本動脈硬化学会総会
  • Place of Presentation: 仙台国際センター（宮城県・仙台市）
  • Year and Date: 2015-07-16
• [Presentation] CETP deficiency in rabbits protects high fat high cholesterol diet induced atherosclerosis (2015)
  • Author(s): Zhang J, Xu J, Liang J, Yang D, Guo Y, Zhu T, Song J, Niimi M, Lai L, Fan L, Chen YE.
  • Organizer: ATVB 2015
  • Place of Presentation: （San Francisco, USA）Hilton San Francisco Union Square Hotel
  • Year and Date: 2015-05-03
  • Int'l Joint Research
• [Book] Fundamentals of Laboratory Animal Science (2017)
  • Author(s): Enqu Liu and Jianglin Fan
  • Total Pages: 352
  • Publisher: CRC Press
• [Remarks] 山梨大学大学院総合研究部医学域基礎医学系分子病理学講座
  • URL: http://www.med.yamanashi.ac.jp/clinical_basic/pathol01/
• [Funded Workshop] International Conference of Rabbit Biotechnology (2017)