Applying a Human Liver Microphysiology System to Develop Therapeutic Strategies for Non-Alcoholic Fatty Liver Disease (NAFLD)

应用人类肝脏微生理学系统制定非酒精性脂肪性肝病 (NAFLD) 的治疗策略

• 批准号: 9920137
• 负责人: D. Lansing Taylor
• 金额: $ 67.45万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-08-01 至 2022-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9920137
• 关键词: 3-Dimensional; Adipose tissue; Animal Model; Animals; Antioxidants; Attenuated; Biological Models; Blood Vessels; Cell model; Cells; Cirrhosis; Clinic; Clinical Data; Complex; Computational Biology; Data; Development; Disease; Disease Pathway; Disease Progression; Disease model; Disease susceptibility; Dose; Drug Combinations; Drug Controls; Drug Screening; Endothelial Cells; Engineering; Environmental Risk Factor; Exhibits; Experimental Models; Fatty Liver; Fibrosis; Gene Mutation; Genes; Genetic; Genetic Diseases; Genetic Variation; Genomics; Genotype; Gluconeogenesis; Goals; Hepatocellular Damage; Hepatocyte; Human; Individual Differences; Inflammation; Inflammatory; Inflammatory Infiltrate; Intestines; Investigation; Kupffer Cells; Lead; Life; Link; Liver; Liver diseases; Measures; Microfluidics; Modeling; Molecular; Molecular Target; Mutation; Oxygen; Pathogenesis; Pathogenicity; Pathway interactions; Patients; Pharmaceutical Preparations; Pharmacologic Substance; Pharmacology; Phenotype; Phospholipase; Physiological; Pioglitazone; Primary carcinoma of the liver cells; Process; Proteins; Reproducibility; Role; SIRT1 gene; Signal Transduction; Single Nucleotide Polymorphism; Statistical Data Interpretation; System; Systems Biology; Technology; Testing; Therapeutic; Time; Tissues; Toxic effect; Variant; Work; adipokines; base; cell type; chronic liver disease; cytokine; disease phenotype; drug candidate; drug testing; efficacy testing; endoplasmic reticulum stress; experience; fatty acid oxidation; functional genomics; genetic signature; hepatic acinus structure; individualized medicine; induced pluripotent stem cell; knock-down; lipid biosynthesis; liver function; liver injury; liver transplantation; loss of function; microphysiology system; new therapeutic target; non-alcoholic fatty liver disease; nonalcoholic steatohepatitis; novel; novel therapeutics; response; stellate cell; success; transcriptome sequencing; urea cycle

We propose to apply four complementary technologies in a Quantitative Systems Pharmacology approach to create a human experimental model of non-alcoholic fatty liver disease (NAFLD), the most rapidly growing disease, and to use the model to test novel therapeutic strategies:1) Implement a vascularized, liver acinus microphysiological system (vLAMPS) constructed with human patient-derived, liver cells, as an experimental model to recapitulate early NAFLD phenotypes and as a platform to experimentally test novel therapeutics; 2) Building on our experience in computational and systems biology, we will use RNAseq data from normal and NAFLD patients to infer pathways of disease progression, to identify the potential molecular protein targets that are in the inferred pathways, and to use our latent factor modeling approach and 3D similarity models to identify drugs that statistically interact with the targets in these pathways; 3) We will employ our highly efficient processes for generating mature iPSC-derived hepatocytes combined with gene editing to incorporate disease engineered iPSC hepatocytes (conditional gain/loss of function) into the vLAMPS to begin testing patient specific therapies; and 4) Apply phenotypic drug screening technologies. NAFLD encompasses a spectrum of liver damage ranging from simple steatosis (NAFL) to more serious non- alcoholic steatohepatitis (NASH), cirrhosis and hepatocellular carcinoma (HCC). Cirrhosis and HCC resulting from progressive damage to the liver have become the third most common causes of liver transplants. The disease pathogenesis of NAFLD is complex and confounded by the considerable inter-individual differences in disease susceptibility, progression and complications, suggesting the need for a patient specific approach. Studies have identified NAFLD associated gene signatures and single nucleotide polymorphisms (SNPs). In particular, the SIRT1 gene that is downregulated in NAFLD, has been identified as a key regulator of lipogenesis, gluconeogenesis, ER stress, fatty acid oxidation, urea cycle and the antioxidant response in hepatocytes. A SNP in the patatin-like phospho-lipase domain-containing 3 (PNPLA3) gene is strongly associated with hepatic steatosis, fibrosis, cirrhosis, and HCC. However, there continues to be major gaps in our understanding of the pathogenesis of NAFLD. For example, despite its strong association with NAFLD, the functional significance of the PNPLA3 variant is unknown. A major limitation in the elucidation of a mechanistic role of PNPLA3 in NAFLD has been the interspecies differences in its expression and tissue-specific distribution, suggesting the need for human cell models. This combination of the technologies and approaches is expected to lead to new strategies for development of repurposed and new therapeutics with the potential to slow or halt the progression of early NAFLD to the more advanced, life threatening stages.

我们建议将四种互补技术应用于定量系统药理学方法 创建人类非酒精性脂肪肝疾病（NAFLD）的实验模型，该模型是最快增长的 疾病，并使用该模型测试新型治疗策略：1） 由人类衍生的肝细胞构建的微生物生理系统（VLAMP）作为实验 概括早期NAFLD表型的模型，并作为实验测试新型治疗剂的平台； 2） 在我们在计算和系统生物学方面的经验的基础上，我们将使用正常和 NAFLD患者推断疾病进展的途径，以确定潜在的分子蛋白靶标的 在推断的途径中，并使用我们的潜在因子建模方法和3D相似性模型 确定在这些途径中与靶标相互作用的药物； 3）我们将利用我们的高效 生成成熟IPSC衍生的肝细胞结合基因编辑以结合疾病的过程 设计的IPSC肝细胞（有条件的增益/功能丧失）进入VLAMP，开始测试患者 具体疗法； 4）应用表型药物筛查技术。 NAFLD包括一系列肝损伤，从简单的脂肪变性（NAFL）到更严重的非 - 酒精性脂肪性肝炎（NASH），肝硬化和肝细胞癌（HCC）。肝硬化和HCC产生 从进行性损害到肝脏已成为肝移植的第三大最常见原因。这 NAFLD的疾病发病机理是复杂的，并且被相当个体间差异所困扰 疾病的敏感性，进展和并发症，表明需要特定于患者的方法。 研究已经确定了NAFLD相关的基因特征和单核苷酸多态性（SNP）。在 特别是，在NAFLD中下调的SIRT1基因已被确定为 脂肪生成，糖异生，ER应激，脂肪酸氧化，尿素周期和抗氧化剂反应 肝细胞。含patatin的磷酸磷酸脂肪酶结构域3（PNPLA3）基因的SNP非常强烈 与肝脂肪变性，纤维化，肝硬化和HCC有关。但是，仍然存在重大差距 我们对NAFLD发病机理的理解。例如，尽管它与NAFLD有很强的联系，但 PNPLA3变体的功能意义尚不清楚。阐明机械的主要限制 PNPLA3在NAFLD中的作用一直是其表达和组织特异性的种间差异 分布，表明需要人类细胞模型。 这些技术和方法的这种结合预计将导致开发的新策略 重新利用和新的治疗剂，有可能减慢或停止早期Nafld的发展 高级，威胁生命的阶段。