Mini-Livers Derived from Human IPS Cells for Modeling Steatosis and Therapy

源自人 IPS 细胞的微型肝脏用于脂肪变性建模和治疗

• 批准号: 10445352
• 负责人: Deepak Nagrath
• 金额: $ 52.65万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-08-01 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10445352
• 关键词: Adult; Alcoholic Liver Cirrhosis; Animals; Cell Communication; Cell Culture Techniques; Cell model; Cells; Cessation of life; Chronic Disease; Cirrhosis; Clinical; Complex; Cues; Data; Deposition; Detection; Development; Diabetes Mellitus; Disease; Engineering; Environment; Epidemic; Epigenetic Process; Evolution; Excision; Fatty Liver; Fatty acid glycerol esters; Fibroblasts; Fibrosis; Funding; Future; Genetic; Genetic Diseases; Genetic Polymorphism; Goals; Hepatic Stellate Cell; Hepatocyte; Heterogeneity; High Prevalence; Histologic; Human; Human Genetics; In Vitro; Inflammatory; Isotopes; Link; Liver; Liver Dysfunction; Liver Fibrosis; Liver diseases; Mediating; Mesenchymal; Metabolic; Metabolic Diseases; Metabolic Pathway; Metabolism; Modeling; Molecular; Non-Insulin-Dependent Diabetes Mellitus; Obesity; Organ; Organ Culture Techniques; Organ Model; Outcome; Pathogenesis; Pathologic; Patients; Pharmacotherapy; Phenotype; Population; Preventive; Primary carcinoma of the liver cells; Process; Public Health; Rattus; Resources; Role; SIRT1 gene; Single Nucleotide Polymorphism; Source; System; Techniques; Technology; Testing; Therapeutic; Therapeutic Intervention; Time; Tissue Model; Tissues; Variant; Work; alternative treatment; base; cell type; chronic liver disease; comorbidity; engineered stem cells; fatty acid biosynthesis; fatty liver disease; genetic variant; high risk; histone modification; human disease; improved; induced pluripotent stem cell; knock-down; lifestyle factors; lipidomics; liver metabolism; macrophage; metabolic profile; metabolomics; non-alcoholic fatty liver disease; nonalcoholic steatohepatitis; novel strategies; prevent; response; small hairpin RNA; stem; stem cell model; stressor; success

ABSTRACT/SUMMARY Our long-term goal remains the same and targets to develop human liver tissue model from stem cells, with multi-cellular cues, metabolic functionality, that incorporates dynamic genetic and epigenetic factors and encompasses the spectrum and evolution of human NAFLD to uncover disease mechanism and therapeutics. The objectives of the proposed study are to continue developing human fatty livers engineered with iPS cells, that incorporates the dynamic expression of the epigenetic-related histone modification SIRT1 in different genetic polymorphisms and elucidate their role as metabolic regulators in the development of human fatty liver disease. The central hypothesis to be tested here is that genetic polymorphisms especially the NAFLD-high- risk PNPLA3 rs738409, combined with epigenetic-related changes (SIRT1) can control hepatic metabolism and contribute to the development of human fatty liver. The rationale is that the ability to generate human diseased liver tissue using genetically modifiable iPS cells from different human populations with single- nucleotide-polymorphisms is a powerful resource, that now make it possible, for the first time, to functionally interrogate their role in disease. Importantly, based on our strong preliminary data of targeted metabolomic analysis in isolated human hepatocytes obtained from either liver resections “Normal Human Hepatocytes” or hepatocytes isolated from explanted NASH livers “NASH Human Hepatocytes” carrying either the NAFLD-high risk PNPLA3 I148M genetic variant or wild type we hypothesize that PNPLA3 I148M hepatocytes have high levels of ferroptosis thereby leading to low metabolic capacity to adapt to stressors and continuous loss of hepatocytes. In the newly proposed aim3, we aim to uncover ferroptosis based therapeutic strategies for patients with NAFLD-high risk PNPLA3 I148M genetic variants. The work described here is expected to i) generate human iPS cells carrying shRNA mediated conditional knockdown of SIRT1 in different genetic backgrounds (especially the NAFLD-high-risk PNPLA3 rs738409 variant), ii) develop a novel approach for modeling an organ-like environment to determine the role of the epigenetic related factor SIRT1 and the genetic variants in the development of fibrotic human mini-livers with fatty liver disease and iii) determine the role of genetic polymorphisms in regulating functional metabolism in mini-human iPS-derived liver tissue with fatty liver disease with especial focus on ferroptosis. Understanding this process alone or in relation to epigenetic changes in a human liver tissue model can result in preventive therapeutic strategies for patients with NAFLD-high risk PNPLA3 I148M genetic variants. The results of this work will also have a positive impact by establishing the basis and platform for future sophisticated organ engineering techniques that incorporates several different cell types from the same iPS cell source, these techniques could be applied to study other liver diseases (e.g. metabolic diseases) and is expected to be a major contribution to the fields of stem cells engineering and liver steatosis.

摘要/摘要 我们的长期目标仍然是相同的，并且是从干细胞开发人肝组织模型的目标 多细胞提示，代谢功能，结合了动态遗传和表观遗传因素以及 涵盖了人类NAFLD在发现疾病机制和治疗方面的频谱和进化。 拟议研究的目标是继续开发由IPS细胞设计的人类脂肪生命， 结合了表观遗传相关的Hisstone修饰SIRT1的动态表达 遗传多态性并阐明其作为代谢调节剂在人类脂肪肝发展中的作用 疾病。这里要检验的中心假设是遗传多态性，尤其是Nafld-High- 风险PNPLA3 RS738409，结合表观遗传相关的变化（SIRT1）可以控制肝脏代谢 并有助于人类脂肪肝的发展。理由是产生人类的能力 患病的肝组织使用来自不同人类种群的普遍可修改的IPS细胞 核苷酸 - 甲状腺真实性是一种强大的资源，现在首次成为可能的资源 询问他们在疾病中的作用。重要的是，基于我们针对性代谢组的强大初步数据 从肝切除术获得的分离的人肝细胞中的分析“正常人肝细胞”或 从外植的纳什生活“纳什人肝细胞”中隔离的肝细胞携带nafld-high 风险PNPLA3 I148M遗传变异或野生型我们假设PNPLA3 I148M肝细胞具有较高 铁凋亡水平，从而导致代谢能力低，以适应压力源和持续丧失 肝细胞。在新提出的AIM3中，我们旨在揭示基于铁铁病的理论策略 NAFLD-HIGH风险PNPLA3 I148M遗传变异的患者。 预计此处描述的工作将i）产生携带shRNA介导的有条件的人IPS细胞 在不同的遗传背景中敲低SIRT1（尤其是NAFLD-HIGH-SHIRK PNPLA3 RS738409 变体），ii）开发一种新颖的方法来建模类似器官的环境来确定 表观遗传学相关因子SIRT1和纤维化人类迷你肝发展中的遗传变异 脂肪肝病和iii）确定遗传多态性在规范功能代谢中的作用 迷你人类IPS衍生的肝组织，具有脂肪肝疾病，特别关注铁铁病。理解 单独或与人肝组织模型中的表观遗传变化有关的过程可能会导致预防 NAFLD-HIGH风险PNPLA3 I148M遗传变异的患者的治疗策略。结果的结果 工作还将通过建立未来复杂器官的基础和平台来产生积极的影响 工程技术结合了来自同一IPS细胞源的几种不同的单元格类型，这些 技术可以应用于研究其他肝病（例如代谢性疾病），预计将是 对干细胞工程和肝脏脂肪变性领域的主要贡献。