Development of a genetic swine model of non-alcoholic steatohepatitis (NASH) by gene-editing

通过基因编辑开发非酒精性脂肪性肝炎（NASH）遗传猪模型

• 批准号: 9410075
• 负责人: Tamene Melkamu
• 金额: $ 39.89万
• 依托单位: RECOMBINETICS, INC.
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-01 至 2018-08-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9410075
• 关键词: Address; Affect; Age; Alcohols; Alleles; Anatomy; Animal Model; Animals; Behavior Therapy; Biochemical; Biotechnology; Blood Chemical Analysis; Body Weight decreased; Breeding; Cardiovascular Diseases; Cell Line; Cells; Cicatrix; Cirrhosis; Clinical; Clinical Chemistry; Cloning; Coronary Arteriosclerosis; Data; Development; Diet; Disease; Economics; Engineering; Enrollment; Environmental Risk Factor; Ethics; Euthanasia; Evaluation; Exhibits; Extrahepatic; FDA approved; Familial Hypercholesterolemia; Family suidae; Fatty acid glycerol esters; Female; Fetus; Fibroblasts; Fibrosis; Founder Generation; Functional disorder; Future; Gene Mutation; Generations; Genes; Genetic; Genetic Risk; Genome; Genotype; Grant; Health; Health Care Costs; Hepatic; Hepatocyte; Hepatology; Histologic; Histopathology; Human; Image; Immune; Immune Evasion; Indiana; Industry; Inflammation; Lead; Link; Liver; Liver Failure; Liver diseases; Longevity; Longitudinal Studies; Mediating; Medical; Metabolic syndrome; Minnesota; Modeling; Morbidity - disease rate; Mutation; Non-Insulin-Dependent Diabetes Mellitus; Nutritional; Obesity; Organ; Oxidative Stress; Pathogenesis; Pathologist; Pathology; Patients; Pharmaceutical Preparations; Pharmacotherapy; Phase; Phenotype; Phospholipase; Physiological; Physiology; Pilot Projects; Primary carcinoma of the liver cells; Proteins; Recording of previous events; Research; Research Personnel; Risk; Severities; Small Business Innovation Research Grant; Technology; Testing; Therapeutic; Therapeutic Intervention; Time; Triglycerides; Universities; Validation; Work; cost; cytokine; diabetic; diagnostic biomarker; falls; fibrogenesis; genetic risk factor; histopathological examination; innovation; interest; lipid metabolism; liver biopsy; liver injury; liver transplantation; male; marker transgenes; mortality; mutant; new therapeutic target; non-alcoholic fatty liver; nonalcoholic steatohepatitis; novel; novel diagnostics; novel therapeutic intervention; novel therapeutics; phase 2 study; pig genome; pre-clinical; prevent; prototype; repaired; success

PROJECT SUMMARY Nonalcoholic fatty liver disease (NAFLD) is the accumulation of fat in liver cells that advance to the more severe form called nonalcoholic steatohepatitis (NASH) characterized by inflammation, scarring (cirrhosis), liver failure and hepatocellular carcinoma, warranting liver transplant. Despite the profound economic and health impacts of NAFLD/NASH, little progress has been made in the management of patients with these severe clinical conditions. While weight loss and anti-diabetic medications are cornerstones of treatment, there is considerable interest in identifying more direct therapeutic interventions. Such developments are currently hampered by lack of an animal model with high physiologic fidelity to the human condition, particularly with respect to the development of fibrosis, immune evasion and metabolic syndrome features that define human NASH. To that end, we propose to utilize our state-of-the-art gene-editing platform to engineer Ossabaw swine with the most potent mutation associated with human NASH, PNPLA3I148M. We hypothesize that Ossabaw pigs carrying PNPLA3I148M alleles will develop histologically severe liver injury typical of human NASH along with metabolic syndrome, with atherogenic dietary challenge. Founder animals will be subject to routine clinical chemistry, lipidomic workups, and histopathological examinations sufficient to qualify fidelity of the model. Success in this endeavor would warrant a Phase II submission wherein the impact of the PNPLA3I148M mutation could be more extensively investigated, namely through more detailed imaging, histologic, cytokine and immune cell phenotyping. In addition to state-of-the art gene editing platform with expertise of Drs. Melkamu (Veterinary Physiology) and Carlson (Animal biotechnology) from Recombinetics, we have engaged as co-investigators, experts in clinical and pathophysiological aspects of NASH in human and in swine nutritional model (Drs. Naga Chalasani and Tiebing Liang from Indiana University) biochemical aspects of NASH , from Vanderbilt University). Drs. Gerry O'Sullivan, a local veterinary pathologist from the University of Minnesota, and Dr. Pierre Bedossa, a world-renown expert in scoring liver histopathology, will serve as consultants. A reliable large animal model of will have tremendous impact on industry and academic research to develop and test new drugs and novel therapeutic approaches to treat this disease. and expertise in (Dr. Charles Robb Flynn NASH

项目概要 非酒精性脂肪性肝病 (NAFLD) 是指肝细胞内脂肪堆积，并进一步发展为脂肪肝。 称为非酒精性脂肪性肝炎 (NASH) 的严重形式，其特征为炎症、疤痕（肝硬化）、肝脏 失败和肝细胞癌，需要肝移植。尽管经济和健康状况深刻 NAFLD/NASH 的影响，这些严重临床患者的治疗进展甚微 状况。虽然减肥和抗糖尿病药物是治疗的基石，但仍有相当多的 有兴趣确定更直接的治疗干预措施。目前，此类发展因缺乏 建立对人类状况具有高度生理保真度的动物模型，特别是在 纤维化、免疫逃避和代谢综合征特征的发展定义了人类 NASH。 为此，我们建议利用我们最先进的基因编辑平台对 Ossabaw 猪进行改造 与人类 NASH 相关的最强突变 PNPLA3I148M。我们假设奥萨博猪携带 PNPLA3I148M 等位基因将导致人类 NASH 典型的组织学严重肝损伤以及代谢 综合征，伴有动脉粥样硬化饮食挑战。创始人动物将接受常规临床化学检查， 脂质组学检查和组织病理学检查足以证明模型的保真度。在这方面取得成功 努力将保证 II 期提交，其中 PNPLA3I148M 突变的影响可能更大 广泛研究，即通过更详细的成像、组织学、细胞因子和免疫细胞 表型分析。 除了具有博士专业知识的最先进的基因编辑平台。 Melkamu（兽医生理学） 和 Recombinetics 的 Carlson（动物生物技术），我们聘请了以下领域的联合研究员和专家： 人类和猪营养模型中 NASH 的临床和病理生理学方面（Naga Chalasani 博士 和来自印第安纳大学的 Tiebing Liang）NASH 的生化方面 ，来自范德比尔特大学）。博士。格里·奥沙利文 (Gerry O'Sullivan)，当地大学兽医病理学家 明尼苏达州和世界著名肝脏组织病理学评分专家 Pierre Bedossa 博士将担任 顾问。一个可靠的大型动物模型将对工业界和学术界产生巨大影响 研究开发和测试治疗这种疾病的新药和新治疗方法。 和专业知识（查尔斯·罗伯博士 弗林 非酒精性脂肪性肝炎