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

描述（由申请人提供）：我们的长期目标是开发具有多细胞因子的天然肝支架，用于干细胞衍生的肝细胞的完全和稳定成熟， 在体外设计功能性肝脏，并将其用于肝脏脂肪变性建模和治疗。该研究的目的是开发一种具有诱导多能干细胞（iPS）衍生肝细胞的肝工程器官培养系统，并研究其用于了解脂肪肝疾病的发病机制，自然史和早期检测工具和治疗方法的发展。这里要检验的中心假设是， 脱细胞的天然肝支架可以广泛地再增殖，将为iPS衍生的肝细胞的代谢成熟提供稳定的器官样环境，并且可以用作使用人野生型iPS细胞或通过敲低SIRT 1和/或（与肝脂肪变性形成有关的关键基因）进行脂肪肝病基因工程后的iPS细胞诱导功能性微型肝形成的方法。这项研究的基本原理是，一旦具有多细胞线索的人类肝脏组织可以在体外可重复地制造出正常和疾病表型，肝脏脂肪变性的发展就可以被操纵，从而产生预防和治疗各种肝脏疾病的新的创新方法。本文描述的工作预计i）产生用于人iPS细胞衍生的肝细胞形成组织的代谢成熟系统，ii）建立携带SIRT 1的shRNA介导的条件性敲低的人iPS细胞，以及iii）开发用于模拟器官样环境以确定SIRT 1在人肝脏脂肪变性或脂肪肝病中的作用的新方法。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建立基础和平台，这些技术结合了几种不同的细胞类型，并可能导致体外整个器官的发育，这些技术可以应用于研究其他肝脏疾病（例如代谢疾病），预计将对干细胞工程和肝脏脂肪变性领域做出重大贡献。

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.