iPS-derived hepatocytes for interrogation of lipid phenotypes

描述（由申请人提供）： 血脂（LDL-C、HDL-C和甘油三酯）的血浆浓度是动脉粥样硬化性心血管疾病的重要危险因素，已被证明是全基因组关联研究（GWAS）的肥沃领域。全球脂质遗传学联盟，一个总计约100，000名受试者的脂质GWAS联盟，已经确定了总共95个基因座，这些基因座在全基因组范围内与至少一种血浆脂质性状显著相关。虽然这个列表包括大多数已知的孟德尔脂质紊乱的原因，大多数是以前没有与脂蛋白代谢。确定这些遗传基因座如何影响与脂质代谢相关的人类组织类型（特别是肝脏）中的表型是关键。这将导致对脂蛋白代谢的生理调节的更好理解，并确定降低LDL-C和TG以及提高HDL-C的新治疗靶点。需要从具有确定基因型的患者中建立无限可再生的功能性肝细胞来源，以研究基因型影响肝细胞脂质生物学和血浆脂质特征的细胞机制。我们已经开发出使用脂肪干细胞（ASC）作为获得重编程诱导多能干细胞（iPS）的基础的能力，并具有致力于ASC分离和扩大iPS生成的核心。通过宾夕法尼亚大学和威斯康星州医学院之间的合作，我们已经建立了将人iPS细胞分化为肝细胞的方案。我们的联盟由iPS生成、肝细胞生成和肝细胞脂质代谢方面的专家组成。我们将开发来自约30000名具有确定基因型的受试者的iPS细胞系和iPS衍生的肝细胞的文库，并将其用于（1）开发高效和可靠的方案以从ASC获得iPS细胞系，随后分化成功能性肝细胞;（2）扩大这些方案以实现iPS细胞系和肝细胞的高通量产生;和（3）从这些肝细胞进行代谢谱分析。这些研究应该提供新的见解的分子机制，其中一些最引人注目的GWAS基因座的血脂性状影响脂蛋白代谢。(End摘要）

DESCRIPTION (provided by applicant): Plasma concentrations of lipids (LDL-C, HDL-C and triglycerides) are important risk factors for atherosclerotic cardiovascular disease and have proven fertile territory for genome-wide association studies (GWAS). The Global Lipids Genetics Consortium, a consortium of lipid GWAS totaling approximately 100,000 subjects, has identified a total of 95 loci that are genome-wide significantly associated with at least one plasma lipid trait. While this list includes most of the genes that are known causes of Mendelian lipid disorders, the majority are not previously associated with lipoprotein metabolism. It is key to determine how these genetic loci affect phenotypes in human tissue types relevant to lipid metabolism, especially the liver. This will lead to a greater understanding of the physiological regulation of lipoprotein metabolism and identify novel therapeutic targets for reducing LDL-C and TG and raising HDL-C. There is a need to establish infinitely renewable sources of functional hepatocytes from patients with defined genotype in order to study the cellular mechanisms by which genotype influences hepatocyte lipid biology and plasma lipid traits. We have developed the ability to use adipose stem cells (ASCs) to serve as the basis for obtaining reprogrammed induced pluripotent stem (iPS) cells and have a core dedicated to both ASC isolation and to scaling up iPS generation. Through collaboration between Penn and Medical College of Wisconsin we have established protocols for the differentiation of human iPS cells into hepatocytes. Our consortium comprises investigators expert in iPS generation, hepatocyte generation, and hepatocyte lipid metabolism. We will develop a library of iPS cell lines and iPS-derived hepatocytes from ~300 hundred subjects with defined genotypes and will use them to (1) develop highly efficient and reliable protocols to obtain iPS cell lines from ASCs, followed by differentiation into functional hepatocytes; (2) scale up these protocols to enable high- throughput generation of iPS cell lines and hepatocytes; and (3) perform metabolic profiling from these hepatocytes. These studies should provide novel insights into the molecular mechanisms by which some of the most compelling GWAS loci for plasma lipid traits influence lipoprotein metabolism. (End of Abstract)