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

基本信息

批准号：
10646093
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
项目类别：
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/10646093
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 Therapy Cell Transplantation Cell physiology Cells Cessation of life Clinical Cytochrome P450 DNA Methylation DNA Methyltransferase Inhibitor Data Development Plans Diabetes Mellitus Diagnosis Differentiated Gene Disease Disease model Elderly Engraftment Epigenetic Process Ethics Exhibits Future Gene Expression Genes Genetic Genus Hippocampus Goals Hepatic Hepatocyte Hepatocyte transplantation Human Human Activities 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 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

项目摘要

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.

项目概要：全球有 8.44 亿人患有肝病，每年死亡人数接近 200 万人。肝移植是某些病例的首选治疗方法，但受到高质量肝移植的限制。来自年轻捐赠者（<55 岁）的器官，以及与手术相关的发病率。使用原代人肝细胞 (PHH) 是治疗特定肝脏疾病的一种微创替代方案结构得以保留但存在代谢紊乱的病理学，例如急性肝功能衰竭细胞的最佳代谢功能对于基于细胞的成功至关重要。然而，PHH 的治疗受到供体稀缺和供体急剧减少的严重限制。来自老年捐献者的 PHH 的代谢功能。诱导多能干细胞 (h-iPSC) 作为 PHH 的替代品出现，用于治疗特定肝脏 H-iHLC 具有三个优点：(1) H-iHLC 由无限的可再生来源生产：h-iPSC。 (2) 它们绕过了与使用胚胎干细胞相关的伦理问题 (3) 它们具有潜力。利用患者自身的细胞来防止移植后的同种异体免疫反应。年龄对 PHH 代谢功能的有害影响已被描述，捐赠者年龄的影响在此，我们的目标是确定与供体年龄相关的变化。通过研究年轻 h-iHLC 的转录组和蛋白质组，了解 h-iHLC 的整体代谢概况和老捐赠者，并将结果与来自相同捐赠者的 PHH 进行比较，我们将详细研究表达。 h-iHLC 和 PHH 中细胞色素 P450 (CYP450) 超家族的功能与供体年龄的关系。与年龄相关的 DNA 甲基化变化会下调 PHH 的代谢功能，包括 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，以优化我们将检查 h-iHLC 中的整体代谢功能，包括 CYP450 活性。通过将 h-iHLC 移植到代谢肝脏中，在急性肝衰竭小鼠模型中生成 h-iHLC 失败，I 型酪氨酸血症（NOD 菌株背景 (FRGN) 上的 Fah´′´/Rag2´´´/Il2rg´´´）小鼠的结果。研究将提供有关捐献者年龄对 h-iHLC 代谢及其调节的影响的重要信息并将 (1) 协助选择代谢适合的供体进行同种异体 h-iHLC 移植，(2) 允许未来通过改变重编程、分化和调节功能和调节机制基因编辑，产生高质量的 h-iHLC，具有针对同种异体和自体基因优化的代谢功能移植。