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: 10623305
• 负责人: Deepak Nagrath
• 金额: $ 52.73万
• 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/10623305
• 关键词: Adult; 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; Disease Progression; 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; Macrophage; 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; Pluripotent Stem Cells; 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; 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; manufacture; 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 我们的长期目标保持不变，目标是从干细胞建立人类肝组织模型， 多细胞信号，代谢功能，结合动态遗传和表观遗传因素 涵盖人类NAFLD的光谱和进化，以揭示疾病机制和治疗方法。 这项拟议研究的目标是继续开发具有诱导性多能性细胞的人类脂肪肝， 它整合了表观遗传相关的组蛋白修饰SIRT1在不同的 基因多态及其作为代谢调节因子在人类脂肪肝发生发展中的作用 疾病。这里要检验的中心假设是遗传多态，特别是NAFLD-高- 风险PNPLA3 rs738409与表观遗传相关变化(SIRT1)相结合可以控制肝脏代谢 并有助于人类脂肪肝的发展。其基本原理是，产生人类的能力 使用来自不同人群的可遗传修改的iPS细胞获得病变的肝组织，这些细胞具有单一的 核苷酸多态是一种强大的资源，它现在第一次使其有可能在功能上 审问他们在疾病中的作用。重要的是，基于我们强大的靶向代谢组初步数据 从“正常人肝细胞”和“正常肝细胞”中分离的人肝细胞的分析 从移植的Nash肝分离的肝细胞“Nash人肝细胞”携带NAFLD-High 风险PNPLA3I148M基因变异或野生型我们假设PNPLA3I148M肝细胞具有高 铁下垂水平，从而导致代谢能力低下，以适应应激源和持续丧失 肝细胞。在新提出的AIM 3中，我们的目标是揭示基于铁性下垂的治疗策略 患有NAFLD高危PNPLA3 I148M基因变异的患者。 本文描述的工作有望：1)产生携带shRNA介导的条件性条件反射的人iPS细胞 SIRT1在不同遗传背景(尤其是NAFLD-高危PNPLA3 rs738409)中的敲除 变种)，二)开发一种新的方法来模拟器官样环境，以确定 表观遗传相关因子SIRT1及其基因变异在人小肝纤维化发生发展中的作用 脂肪肝病和III)确定基因多态在调节功能代谢中的作用 脂肪肝病患者的迷你人类iPS来源的肝组织，特别关注铁性下垂。理解 这一过程单独或与人类肝组织模型中的表观遗传变化有关，可以导致预防性的 NAFLD高危PNPLA3I148M基因变异患者的治疗策略。这样做的结果 工作也将产生积极的影响，为未来复杂的器官建立基础和平台 工程技术结合了来自相同iPS细胞来源的几种不同类型的细胞，这些 技术可以应用于研究其他肝病(如代谢性疾病)，并有望成为 在干细胞工程和肝脏脂肪变性领域做出了重大贡献。

项目成果

Future Economics of Liver Transplantation: A 20-Year Cost Modeling Forecast and the Prospect of Bioengineering Autologous Liver Grafts.

• DOI: 10.1371/journal.pone.0131764
• 发表时间: 2015
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Habka D;Mann D;Landes R;Soto-Gutierrez A
• 通讯作者: Soto-Gutierrez A

Biotechnology Challenges to In Vitro Maturation of Hepatic Stem Cells.

• DOI: 10.1053/j.gastro.2018.01.066
• 发表时间: 2018-04
• 期刊: Gastroenterology
• 影响因子: 29.4
• 作者: Chen C;Soto-Gutierrez A;Baptista PM;Spee B
• 通讯作者: Spee B

Biofabrication of Autologous Human Hepatocytes for Transplantation: How Do We Get There?

用于移植的自体人肝细胞的生物制造：我们如何实现这一目标？

• DOI: 10.3727/105221618x15350366478989
• 发表时间: 2019
• 期刊: Gene expression
• 作者: Agarwal,Nandini;Popovic,Branimir;Martucci,NicoleJ;Fraunhoffer,NicolasA;Soto-Gutierrez,Alejandro
• 通讯作者: Soto-Gutierrez,Alejandro

Liver-Regenerative Transplantation: Regrow and Reset.

• DOI: 10.1111/ajt.13678
• 发表时间: 2016-06
• 期刊: American journal of transplantation : official journal of the American Society of Transplantation and the American Society of Transplant Surgeons
• 作者: Collin de l'Hortet A;Takeishi K;Guzman-Lepe J;Handa K;Matsubara K;Fukumitsu K;Dorko K;Presnell SC;Yagi H;Soto-Gutierrez A
• 通讯作者: Soto-Gutierrez A

Clinical Hepatocyte Transplantation: What Is Next?

• DOI: 10.1007/s40472-017-0165-6
• 发表时间: 2017-12
• 期刊: Current transplantation reports
• 影响因子: 2.1
• 作者: Squires JE;Soltys KA;McKiernan P;Squires RH;Strom SC;Fox IJ;Soto-Gutierrez A
• 通讯作者: Soto-Gutierrez A