The effect of donor age on the function and therapeutic efficacy of human hepatocyte-like cells

供者年龄对人肝细胞样细胞功能及治疗效果的影响

• 批准号: 10674009
• 负责人: Varvara A Kirchner
• 金额: $ 16.51万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-03-15 至 2026-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10674009
• 关键词: Acute Liver Failure; Address; Adult; Affect; Age; Age Years; Allogenic; Animals; Architecture; Attention; Autologous; Bilirubin; Biological Assay; Biology of Aging; Bypass; Cell Differentiation process; Cell Maturation; Cell Reprogramming; Cell Therapy; Cell Transplantation; Cell physiology; Cells; Cessation of life; Clinical; Cytochrome P450; DNA Methylation; DNA Methyltransferase Inhibitor; Data; Data Correlations; Development Plans; Diabetes Mellitus; Diagnosis; Disease; Disease model; Education; Elderly; Engraftment; Epigenetic Process; Ethics; Exhibits; Future; Gene Expression; Genes; Genetic; Genus Hippocampus; Goals; Hepatic; Hepatocyte; Hepatocyte transplantation; Human; Immune response; In Vitro; Liver; Liver Failure; Liver diseases; Macular degeneration; Mass Spectrum Analysis; Measures; Mentors; Metabolic; Metabolism; Monitor; Morbidity - disease rate; Mus; Operative Surgical Procedures; Organ; Organ Transplantation; Oxygen Consumption; Pathology; Patients; Persons; Pharmacy (field); Play; Positioning Attribute; Proteins; Proteome; Protocols documentation; Publishing; Regenerative Medicine; Regulation; Rejuvenation; Research Personnel; Risk Factors; Role; Serum Albumin; Solid; Source; Technology; Testing; Training; Transaminases; Transcript; Transplant Surgeon; Transplantation; Treatment Efficacy; Tyrosinemias; age group; age related; aged; alternative treatment; career; career development; cell age; clinical application; embryonic stem cell; epigenetic regulation; epigenome; experience; hepatocyte engraftment; in vivo; induced pluripotent stem cell; liver transplantation; metabolic profile; minimally invasive; mortality; mouse model; multiple omics; novel; operation; preservation; prevent; stem cell biology; stem cells; success; therapeutic evaluation; transcriptome; transcriptome sequencing

Project Summary: Worldwide, 844 million people are afflicted with liver disease, with mortality nearing 2 million deaths per year. Liver transplantation is the preferred treatment for selected cases but is limited by the availability of high-quality organs from young donors (<55 years old), and the morbidity associated with the operation. Cell-based therapy using primary human hepatocytes (PHHs) is a minimally invasive alternative for treatment of select liver pathologies where architecture is preserved but there are metabolic derangements, such as in acute liver failure and metabolic liver disease. Optimal metabolic function of the cells is critical for the success of cell-based therapies. However, PHH therapy is severely limited by the scarcity of donors, and the dramatic decrease in metabolic function of PHHs from older donors. Human hepatocyte-like cells (h-iHLCs) derived from human induced pluripotent stem cells (h-iPSCs) emerged as an alternative to PHHs for treatment of select liver conditions. H-iHLC have three benefits: (1) H-iHLCs are produced from an unlimited, renewable source: h-iPSCs. (2) They bypass ethical concerns associated with the use of embryonic stem cells. (3) They have the potential to prevent an allogeneic immune response following transplantation by utilizing the patients’ own cells. Although, the deleterious impact of age on the metabolic function has been described for PHHs, the impact of donor age on the metabolism in h-iHLCs has not been studied. Here, we aim to identify the donor age-associated changes in the overall metabolic profile of h-iHLCs by studying the transcriptome and proteome of h-iHLCs from young and old donors and compare the results to PHHs from the same donors. We will study in detail the expression and function of the cytochrome P450 (CYP450) superfamily in h-iHLCs and PHHs as a function of donor age. Age-related changes in DNA-methylation down-regulate metabolic function including CYP450 activity in PHHs. Therefore, we will study and attempt to modulate this regulatory mechanism in h-iHLCs with the goal to optimize the overall metabolic function including CYP450 activity in h-iHLCs. We will examine the therapeutic efficacy of the generated h-iHLCs in a murine model of acute liver failure by transplanting h-iHLCs into metabolic liver failure, tyrosemia type I (Fah¯′¯/Rag2¯′¯/Il2rg¯′¯ on NOD-strain background (FRGN)) mice. The results from this study will provide critical information about the impact of donor age on metabolism and its regulation in h-iHLCs and will (1) assist in selecting metabolically fit donors for allogeneic h-iHLCs transplantation, (2) allow future modulation of functional and regulatory mechanisms through alterations in reprogramming, differentiation and gene editing, to produce high-quality h-iHLCs with optimized metabolic function for allo- and autogeneic transplantation.

项目摘要： 在全球范围内，有8.44亿人患有肝病，死亡率接近200万人死亡。 肝移植是选定病例的首选治疗方法，但受高质量可用性的限制 来自年轻捐助者的器官（<55岁），以及与手术有关的发病率。基于细胞的疗法 使用原发性人肝细胞（PHHS）是一种用于治疗精选肝脏的微创替代品 保留结构但存在代谢发展的病理，例如急性肝衰竭 和代谢性肝病。细胞的最佳代谢功能对于基于细胞的成功至关重要 疗法。但是，PHH疗法受到捐助者的稀缺性严重限制，并且急剧下降 老年捐助者的PHH的代谢功能。源自人的人肝细胞样细胞（H-IHLC） 诱导多能干细胞（H-IPSC）作为PHHS的替代品以治疗精选肝脏 状况。 H-IHLC具有三个好处：（1）H-IHLC是从无限的可再生资源的：H-ipscs产生的。 （2）它们绕过与胚胎干细胞使用有关的道德问题。 （3）他们有潜力 为了防止使用患者自己的细胞移植后同种异体免疫反应。虽然， 年龄对代谢功能的删除影响已被描述为PHHS，供体年龄的影响 H-IHLC中的新陈代谢尚未研究。在这里，我们旨在确定与年龄相关的变化 在H-IHLC的整体代谢谱中，通过研究Young的H-IHLC的转录组和蛋白质组 和旧的捐助者，并将结果与​​来自同一捐助者的PHH进行比较。我们将详细研究表达 H-IHLC和PHHS中的细胞色素P450（CYP450）的功能与供体年龄的函数。 DNA-甲基化与年龄相关的变化下调代谢功能，包括PHHS中的CYP450活性。 因此，我们将研究并尝试调节H-IHLC中的这种调节机制，以优化 总体代谢功能包括H-IHLC中的CYP450活性。我们将检查 通过将H-IHLC移植到代谢性肝脏中，在急性肝衰竭的鼠模型中生成的H-IHLC 失败，I型（fah''/rag2''/il2rg''在点头型背景（FRGN））小鼠。从中的结果 研究将提供有关供体年龄对新陈代谢及其在H-IHLC中的调节的影响的关键信息 并将（1）协助选择代谢拟合的供体进行同种异体H-IHLC的移植，（2）允许将来 通过改变重编程，分化和调节功能和调节机制的调节 基因编辑，生产具有优化代谢功能的高质量H-IHLC 移植。