Ex-vivo bioengineered technology to unravel dysfunction due to non-alcoholic steatohepatitis (NASH)

离体生物工程技术可解决非酒精性脂肪性肝炎 (NASH) 引起的功能障碍

• 批准号: 10744393
• 负责人: Richard M Green
• 金额: $ 70.05万
• 依托单位: UNIVERSITY OF ARIZONA
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-01 至 2027-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10744393
• 关键词: Agonist; American; Animal Model; Animals; Bile Acids; Bioinformatics; Biological Markers; Biology; Biomedical Engineering; Cell Differentiation process; Cell Line; Cell Transplantation; Cells; Cellular Metabolic Process; Cellular Stress; Cessation of life; Characteristics; Chemicals; Cirrhosis; Clinical; Collaborations; Development; Diabetes Mellitus; Diet; Disease Progression; Drug Screening; Environment; Environmental Risk Factor; Evaluation; Experimental Designs; Extracellular Matrix; Fatty Acids; Fibrosis; Functional disorder; Gel; Gene Expression; Growth; Hepatic; Hepatocyte; Hepatology; Human; Individual; Institution; Insulin Resistance; Investigation; Lipids; Liver; Liver Failure; Liver Fibrosis; Liver Function Tests; Liver diseases; Measures; Metabolic; Metabolic Pathway; Metabolic stress; Metabolic syndrome; Molecular Chaperones; Obesity; Pathogenesis; Pathogenicity; Pathway interactions; Patients; Peripheral Blood Mononuclear Cell; Persons; Pharmaceutical Preparations; Phenotype; Primary carcinoma of the liver cells; Proteins; Recurrence; Recurrent disease; Reducing Agents; Risk; Severity of illness; Signal Transduction; System; Technology; Testing; Transplantation Surgery; United States; Virulence Factors; bile acid metabolism; biomarker development; cell growth; cell type; cost; disorder control; disorder risk; drug development; endoplasmic reticulum stress; experience; genetic risk factor; high throughput technology; high-throughput drug screening; human model; in vivo; induced pluripotent stem cell; lipid metabolism; lipidomics; liver biopsy; liver injury; liver stiffness; liver transplantation; metabolomics; minimally invasive; non-alcoholic fatty liver disease; nonalcoholic steatohepatitis; novel; pharmacologic; post-transplant; precision drugs; resistance factors; response; stem cell biology; stem cells; stress reduction; tauroursodeoxycholic acid; tool; trait

SUMMARY Non-alcoholic fatty liver disease (NAFLD) is the most common cause of abnormal liver function tests in the US and its progressive form, termed non-alcoholic steatohepatitis (NASH), will soon be the leading indication for liver transplantation. There are currently no effective medications to treat NASH, no biomarkers to determine disease progression or risk of post-transplant recurrence and no effective platforms for high-throughput drug screening. Although NASH is related to obesity and diabetes, the pathogenic factors that cause disease progression to NASH/Cirrhosis are poorly understood. Compared to an invasive liver biopsy, peripheral blood mononuclear cells (PBMCs) can be easily obtained from patients with NASH and end-stage NASH/Cirrhosis patients requiring liver transplantation and re-programmed into induced pluripotent stem cells (iPSCs). These iPSCs may then be differentiated into iPSC-hepatocytes, which are human liver-like cells that can be cultured in ex vivo bioengineered systems tailored to normal and cirrhotic liver stiffness, enabling an investigation that is independent of the compounding metabolic and environmental factors that complicate analysis within human or animal systems. In this proposal, we will utilize an iPSC-hepatocyte platform to determine the effects of extracellular matrix (ECM) stiffness and unfolded protein response (UPR) cell signaling on hepatic lipid metabolism. We will initially unwind the impact and interplay between matrix stiffness and patient-specific propensity for NASH in patient-derived iPSCs and analyze the impact on lipid metabolism and lipidomics (Aim 1). ER stress and the unfolded protein response (UPR) has been shown to be important in the pathogenesis of NASH. Thus, we will use iPSC-hepatocytes to develop a platform for determining the interaction between UPR signaling and lipid metabolism relevant to NASH (Aim 2). iPSC-hepatocytes will be treated with ER stress reducing compounds including the chemical chaperone tauroursodeoxycholic acid (TUDC) or FXR/bile acid agonists, and the effects on cell differentiation, gene expression and lipid metabolism will investigated. Finally, iPSC-hepatocytes will be used to study the cell signaling and pathogenic mechanisms of NASH in iPSCs from patients with rapidly progressive NASH/Cirrhosis that require liver transplantation. We will develop an iPSC- hepatocyte platform identifying matrix and UPR factors responsible for NASH using iPSC-hepatocytes from NASH/Cirrhosis patients listed for liver transplantation (Aim 3). This MPI proposal leverages the collaboration between three PIs at two institutions with extensive experience investigating 1) iPSCs, bioengineering matrices, ECM biology, and transplant surgery, 2) hepatic lipid metabolism, cell signaling and transplant hepatology, and 3) lipidomics and metabolomics. The development and optimization of these iPSC-hepatocyte platforms will have important implications for determining the pathophysiology of NASH, developing biomarkers to determine risk for NASH progression and for use in drug development and personalized drug screening.

概括 非酒精性脂肪肝 (NAFLD) 是美国肝功能检查异常的最常见原因 其进行性形式，称为非酒精性脂肪性肝炎 (NASH)，很快将成为以下疾病的主要适应症： 肝移植。目前尚无治疗 NASH 的有效药物，也没有可确定的生物标志物 疾病进展或移植后复发的风险，并且没有有效的高通量药物平台 筛选。虽然NASH与肥胖和糖尿病有关，但引起疾病的致病因素 NASH/肝硬化的进展尚不清楚。与侵入性肝活检相比，外周血 单核细胞 (PBMC) 可轻松从 NASH 和终末期 NASH/肝硬化患者中获取 需要肝移植并重新编程为诱导多能干细胞（iPSC）的患者。这些 然后，iPSC 可能会分化为 iPSC 肝细胞，这是一种类似人肝的细胞，可以在 针对正常和肝硬化肝硬度量身定制的离体生物工程系统，使研究成为可能 独立于复杂的代谢和环境因素，这些因素使人体或体内的分析变得复杂 动物系统。在本提案中，我们将利用 iPSC-肝细胞平台来确定 肝脂质上的细胞外基质 (ECM) 硬度和未折叠蛋白反应 (UPR) 细胞信号传导 代谢。我们首先将消除基质刚度和患者特异性之间的影响和相互作用 患者来源的 iPSC 中 NASH 的倾向，并分析其对脂质代谢和脂质组学的影响（Aim 1）。 ER 应激和未折叠蛋白反应 (UPR) 已被证明在以下疾病的发病机制中很重要 纳什。因此，我们将使用 iPSC-肝细胞开发一个平台来确定 UPR 之间的相互作用 与 NASH 相关的信号传导和脂质代谢（目标 2）。 iPSC-肝细胞将接受 ER 应激处理 还原化合物，包括化学伴侣牛磺熊去氧胆酸 (TUDC) 或 FXR/胆汁酸 激动剂，并将研究对细胞分化、基因表达和脂质代谢的影响。最后， iPSC-肝细胞将用于研究 iPSC 中 NASH 的细胞信号传导和致病机制 患有快速进展的 NASH/肝硬化且需要肝移植的患者。我们将开发 iPSC- 肝细胞平台使用 iPSC 肝细胞识别导致 NASH 的基质和 UPR 因子 NASH/肝硬化患者被列入肝移植名单（目标 3）。该 MPI 提案利用了协作 两个机构的三位 PI 之间的合作，他们具有丰富的调查经验：1) iPSC、生物工程矩阵、 ECM 生物学和移植手术，2) 肝脂质代谢、细胞信号传导和移植肝病学，以及 3）脂质组学和代谢组学。这些 iPSC-肝细胞平台的开发和优化将具有 对于确定 NASH 的病理生理学、开发生物标志物来确定风险具有重要意义 用于 NASH 进展以及用于药物开发和个性化药物筛选。