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: 9329289
• 负责人: Alejandro Soto-Gutierrez
• 金额: $ 33.5万
• 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 2018-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9329289
• 关键词: Animal Model; Bile Acids; Bile Duct Epithelium; Blood Vessels; Cadaver; Cell Communication; Cell Differentiation process; Cell model; Cells; Cessation of life; Cirrhosis; Cues; Cultured Cells; Cytochrome P450; Data; Deposition; Detection; Development; Disease; Early Diagnosis; Elements; Embryo; Employment; Engineering; Environment; Epithelial Cells; Evolution; Exhibits; Fatty Acids; Fatty Liver; Fatty acid glycerol esters; Fibrosis; Future; Genes; Genetic Engineering; Genetic Transcription; Glycolysis; Goals; Hepatic; Hepatic Tissue; Hepatocyte; Homeostasis; Human; In Vitro; Inflammation; Inflammatory; Insulin Resistance; Lead; Liver; Liver Dysfunction; Liver diseases; Mediating; Messenger RNA; Metabolic; Metabolic Diseases; Metabolism; Mitochondria; Modeling; Molecular; Mus; Natural History; Organ; Organ Culture Techniques; Organ Model; Pathogenesis; Pathologic; Pathway interactions; Pharmacology; Phenotype; Prevention approach; Primary carcinoma of the liver cells; Production; Proteins; Protocols documentation; Public Health; Research; Role; SIRT1 gene; Stem cells; System; Tamoxifen; Techniques; Technology; Testing; Therapeutic; Time; Tissue Model; Tissues; Transplantation; Triglycerides; Work; alternative treatment; cell type; cellular engineering; disease phenotype; improved; induced pluripotent stem cell; innovation; knock-down; lipid metabolism; liver development; liver function; liver metabolism; liver transplantation; macrophage; metabolic engineering; nonalcoholic steatohepatitis; novel strategies; prevent; public health relevance; scaffold; small hairpin RNA; stem; stem cell differentiation; success; three dimensional structure; tool

DESCRIPTION (provided by applicant): Our long-term goal is to develop a natural hepatic scaffold with multi-cellular cues for complete and stable maturation of stem-derived liver cells to engineer functional livers in vitro and use them for modeling liver steatosis and therapeutics. The objectives of the proposed study are to develop an organ culture system for liver engineering with induced pluripotent stem (iPS) cell-derived liver cells, and investigate its employment to understand pathogenesis, natural history and development of early detection tools and treatments for fatty liver diseases. The central hypothesis to be tested here is that the decellularized natural liver scaffold can be extensively repopulated, will provide a stable organ-like environment for the metabolic maturation of iPS derived liver cells, and may be used as an approach to induce formation of functional mini-livers using human wild type iPS cells or iPS cells after genetic engineer for fatty liver disease by knockdown of SIRT1 and/or (key gene implicated with liver steatosis formation). The rationale for the proposed research is that, once human liver tissue with multi-cellular cues can be reproducibly manufactured in vitro with normal and disease phenotypes, development of liver steatosis can be manipulated pharmacologically, resulting in new and innovative approaches to the prevention and treatment of a variety liver disease. The work described here is expected to i) generate a metabolic maturation system for human iPS cell-derived liver cells to form tissue, ii) establish human iPS cells carrying shRNA mediated conditional knockdown of SIRT1 and iii) develop a novel approach for modeling an organ-like environment to determine the role of SIRT1 in human liver steatosis or fatty liver disease. 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 and may lead to development of entire organs in vitro, 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.

描述(申请人提供)：我们的长期目标是开发一种具有多细胞线索的天然肝支架，以使干细胞完全和稳定地成熟到 在体外设计功能性肝脏，并将其用于模拟肝脏脂肪变性和治疗。本研究的目的是利用诱导多能干细胞(IPS)来源的肝细胞建立肝脏工程的器官培养系统，并探讨其在了解脂肪肝疾病的发病机制、自然病程以及早期检测工具和治疗方法方面的应用。这里要检验的中心假设是， 脱细胞天然肝支架可广泛再生，将为iPS源性肝细胞的代谢成熟提供稳定的器官样环境，并可用于通过敲除SIRT1和/或(参与肝脏脂肪变性形成的关键基因)基因工程治疗脂肪肝后，利用人野生型iPS细胞或iPS细胞诱导形成功能性微型肝脏。这项拟议研究的基本原理是，一旦具有多细胞提示的人类肝组织能够在体外以正常和疾病表型重复制造，就可以通过药物操纵肝脏脂肪变性的发展，从而产生预防和治疗各种肝脏疾病的新的创新方法。本文所描述的工作有望：1)建立人iPS细胞来源的肝细胞形成组织的代谢成熟系统；2)建立携带shRNA介导的SIRT1条件性敲除的人iPS细胞；3)发展一种新的方法来模拟器官样环境，以确定SIRT1在人类肝脏脂肪变性或脂肪肝中的作用。这项工作的结果也将产生积极的影响，为未来复杂的器官工程技术奠定基础和平台，这些技术结合了几种不同的细胞类型，可能导致整个器官的体外发育，这些技术可以应用于其他肝脏疾病(如代谢性疾病)的研究，有望对干细胞工程和肝脏脂肪变性领域做出重大贡献。