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.

我们建议在定量系统药理学方法中应用四种互补技术 创建非酒精性脂肪性肝病 (NAFLD) 的人体实验模型，NAFLD 是增长最快的疾病 疾病，并使用该模型来测试新的治疗策略：1）实施血管化肝腺泡 微生理系统（vLAMPS）由人类患者来源的肝细胞构建，作为实验 重现早期 NAFLD 表型的模型，并作为实验测试新疗法的平台； 2） 基于我们在计算和系统生物学方面的经验，我们将使用来自正常和系统生物学的 RNAseq 数据。 NAFLD 患者推断疾病进展的途径，识别潜在的分子蛋白靶标 位于推断的路径中，并使用我们的潜在因素建模方法和 3D 相似性模型 确定与这些途径中的靶点在统计上相互作用的药物； 3）我们将利用我们的高效 生成成熟 iPSC 衍生肝细胞的过程与基因编辑相结合以纳入疾病 将 iPSC 肝细胞（有条件获得/丧失功能）工程化至 vLAMPS 中以开始测试患者 具体疗法； 4）应用表型药物筛选技术。 NAFLD 涵盖一系列肝损伤，从简单的脂肪变性 (NAFL) 到更严重的非脂肪性肝损伤。 酒精性脂肪肝炎（NASH）、肝硬化和肝细胞癌（HCC）。导致肝硬化和肝癌 肝脏的进行性损伤已成为肝移植的第三大常见原因。这 NAFLD 的发病机制非常复杂，并且由于个体间的巨大差异而令人困惑。 疾病的易感性、进展和并发症，表明需要针对患者的具体方法。 研究已经确定了 NAFLD 相关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 NAFLD 的关键调节因子。 脂肪生成、糖异生、内质网应激、脂肪酸氧化、尿素循环和抗氧化反应 肝细胞。含马铃薯蛋白样磷脂酶结构域 3 (PNPLA3) 基因中的 S​​NP 强烈 与肝脂肪变性、纤维化、肝硬化和 HCC 相关。然而，仍然存在重大差距 我们对 NAFLD 发病机制的了解。例如，尽管它与 NAFLD 密切相关， PNPLA3 变体的功能意义尚不清楚。阐明机械原理的主要局限性 PNPLA3 在 NAFLD 中的作用在于其表达的种间差异和组织特异性 分布，表明需要人类细胞模型。 技术和方法的结合预计将带来新的发展战略 重新调整用途的新疗法有可能减缓或阻止早期 NAFLD 进展为更严重的疾病 晚期、危及生命的阶段。