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

项目摘要 非酒精性脂肪性肝病（NAFLD）是脂肪在肝细胞中的积累， 称为非酒精性脂肪性肝炎（NASH）的严重形式，其特征在于炎症、瘢痕形成（肝硬化）、肝硬化、肝硬化和肝硬化。 失败和肝细胞癌，阻止肝移植。尽管经济和健康状况 由于NAFLD/NASH的影响，在这些严重临床疾病患者的管理方面几乎没有取得进展。 条件虽然减肥和抗糖尿病药物是治疗的基石，但 有兴趣确定更直接的治疗干预措施。这些发展目前受到缺乏 对人类状况具有高度生理保真度的动物模型，特别是关于 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来工程化Ossabaw猪， 与人类NASH相关的最有效的突变，PNPLA 3 I148 M。我们假设奥萨鲍猪携带 PNPLA 3 I 148 M等位基因将发展人类NASH典型的组织学上严重的肝损伤沿着代谢性肝损伤。 综合征，伴有致动脉粥样硬化饮食挑战。将对创始动物进行常规临床化学检查， 脂质组学检查和组织病理学检查足以鉴定模型的保真度。胜任这 奋进将保证II期提交，其中PNPLA 3 I148 M突变的影响可能更多 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 除了拥有Melkamu博士（兽医生理学）专业知识的最先进的基因编辑平台外， 和Carlson（动物生物技术），我们聘请了合作研究者， NASH在人类和猪营养模型中的临床和病理生理学方面（那牙Chalasani博士 和来自印第安纳州大学的TiebingLiang）NASH的生物化学方面 来自范德比尔特大学）。格里·奥沙利文博士，来自纽约大学的当地兽医病理学家， 明尼苏达州，和皮埃尔Bedossa博士，一个世界著名的专家在评分肝脏组织病理学，将担任 顾问。可靠的大型动物模型将对工业和学术产生巨大的影响 研究开发和测试新的药物和新的治疗方法来治疗这种疾病。 查尔斯·罗伯博士（Dr. Charles Robb） 弗林 纳什

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