New approaches to differentiation of human stem cells into hepatocytes

人类干细胞分化为肝细胞的新方法

• 批准号: 7960926
• 负责人: James Lue
• 金额: $ 15.39万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-30 至 2015-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7960926
• 关键词: Address; Adipose tissue; Adult; Animal Model; Animals; Antibodies; Applications Grants; Area; Award; Bone Marrow; CXCR4 Receptors; CXCR4 gene; Cell Nucleus; Cell Therapy; Cell Transplantation; Cell Transplants; Cells; Chemotaxis; Childhood; Chromatin; Clinical; Complex; Data; Defect; Derivation procedure; Development; Development Plans; Developmental Biology; Embryo; Endoderm; Engraftment; Fellowship; Fibroblasts; G-Protein-Coupled Receptors; Gastroenterology; Gene Expression; Generations; Genetic; Genome; Genomics; Germ Layers; Goals; Hand; Hematopoietic stem cells; Hepatic; Hepatitis C virus; Hepatocyte; Homing; Hour; Human; Human Engineering; Immunodeficient Mouse; In Vitro; Infusion procedures; Injury; Investigation; Life; Liver; Liver Failure; Liver Regeneration; Liver Stem Cell; Liver diseases; Mentors; Mentorship; Mesenchymal; Mesenchymal Stem Cells; Metabolic; Methods; Modeling; Molecular; Molecular Virology; Mus; Neoplasm Metastasis; Organ; Organ Donor; Organ Transplantation; Patients; Pediatrics; Physicians; Play; Population; Reporting; Research; Research Personnel; Rodent Model; Role; Safety; Scientist; Signal Pathway; Signal Transduction; Solutions; Source; Spleen; Stem cell transplant; Stem cells; Stromal Cell-Derived Factor 1; Supplementation; System; Techniques; Tissues; Training; Translating; Transplantation; United Network for Organ Sharing; United States; Universities; Up-Regulation; Waiting Lists; adult stem cell; authority; base; career; career development; cell motility; cell type; drug metabolism; embryonic stem cell; gastrulation; grasp; hepatocyte engraftment; human embryonic stem cell; human stem cells; improved; improved functioning; induced pluripotent stem cell; interest; knowledge base; liver function; liver transplantation; member; migration; mouse model; novel; novel strategies; overexpression; programs; public health relevance; receptor; response; skills; statistics; stem cell biology; stem cell population; stem cell therapy; trafficking; transcription factor

DESCRIPTION (provided by applicant): This is an application for a K08 award that outlines a 5-year training plan to facilitate Dr. James Lue's career development as an independent academic physician-scientist. Dr. Lue recently completed his clinical fellowship in pediatric gastroenterology at Stanford University and is establishing himself as a young investigator in stem cell biology research. This K08 award will provide Dr. Lue with the support necessary to accomplish the following training goals: (1) to become an expert in liver stem cell biology, (2) to become an expert in advanced techniques in transcriptional profiling and gene expression analysis, (3) to become an expert in manipulation of animal models for the study of hepatocyte and stem cell transplantation, and (4) to develop an independent physician-scientist career. To achieve these goals, Dr. Lue has assembled a mentoring team comprised of a primary mentor, Dr. Jeffrey Glenn, who is an internationally recognized authority on the molecular virology of hepatitis C virus (HCV), and has additional interests in the differentiation of liver stem cells, human hepatocyte repopulation in small animals, and engineered human liver tissues, and three co-mentors: Dr. Julie Baker, who specializes in the genomics of human definitive endoderm and the generation of hepatocytes from human embryonic stem cells; Dr. Gary Peltz, who specializes in the development of genetic mouse models for the study of human liver disease and drug metabolism; and Dr. Eric Sibley, who is a member of the Pediatrics Mentoring Program at Stanford University and will provide academic career advice and mentorship throughout the award period. Due to the vast shortage of organ donors, researchers have long been in search of alternatives to whole-liver transplantation. Therapies involving repopulation of the liver via cell transplantation have been evolving over the past two decades. Dr. Lue's long-term objective is to produce large numbers of functioning hepatocytes from stem cells so that therapies can be developed to treat patients with liver failure. This project will focus on improving the efficiency of derivation of hepatocytes from human embryonic stem cells (Aim 1) and adult mesenchymal stem cells (Aim 2). Dr. Lue plans to improve differentiation efficiency by priming these stem cells with specific transcription factors (TFs) that are important in normal liver development. Overexpression of these TFs will produce a more homogeneous and competent stem cell population that is more suitable for subsequent hepatic differentiation. Aim 3 will focus on hepatocyte transplantation and improving hepatocyte engraftment by exploiting a well-described homing system that is native to the liver. Fresh human hepatocytes, as well as the stem cell-derived hepatocytes from Aims 1 & 2, will be modified to overexpress a specific homing receptor and intrasplenically injected into highly immunodeficient mice to study liver engraftment and function. This research will form the basis for an R01 grant application conducting research on novel methods of developing clinically useful hepatocytes from stem cells. PUBLIC HEALTH RELEVANCE: Close to 16,000 people are currently on the waiting list for a liver transplant in the United States, the demand for organs is more than double the supply, and over 1,500 people die each year on the waiting list (statistics from the United Network for Organ Sharing, unos.org). Stem cell therapy for the treatment of liver disease is a promising alternative to whole-organ transplantation, but still requires further research to improve safety and efficacy.

描述（由申请人提供）： 这是一份K 08奖项的申请，概述了一个5年的培训计划，以促进James Lue博士作为一名独立的学术物理学家和科学家的职业发展。Lue博士最近完成了他在斯坦福大学的儿科胃肠病学临床研究，并正在成为干细胞生物学研究的年轻研究者。该K 08奖项将为Lue博士提供必要的支持，以实现以下培训目标：（1）成为肝干细胞生物学专家，（2）成为转录谱和基因表达分析先进技术专家，（3）成为肝细胞和干细胞移植研究动物模型操作专家，（4）发展独立的医生-科学家职业。为了实现这些目标，Lue博士组建了一个指导团队，由主要导师Jeffrey Glenn博士和三位共同导师组成，Jeffrey Glenn博士是国际公认的丙型肝炎病毒（HCV）分子病毒学权威，并对肝脏干细胞分化，小动物中的人类肝细胞再生和工程化人类肝脏组织有额外的兴趣：Julie Baker博士，专门从事人类定形内胚层的基因组学和从人类胚胎干细胞产生肝细胞;加里佩尔茨博士，专门从事人类肝病和药物代谢研究的遗传小鼠模型的开发;埃里克·西布利博士是斯坦福大学儿科导师计划的成员，他将在整个颁奖期间提供学术职业建议和导师。 由于器官捐献者的严重短缺，研究人员长期以来一直在寻找全肝移植的替代方案。在过去的二十年中，涉及通过细胞移植使肝脏再生的疗法一直在发展。Lue博士的长期目标是从干细胞中产生大量功能性肝细胞，以便开发治疗肝功能衰竭患者的疗法。该项目将致力于提高从人胚胎干细胞（Aim 1）和成人间充质干细胞（Aim 2）中获得肝细胞的效率。Lue博士计划通过用在正常肝脏发育中重要的特异性转录因子（TF）引发这些干细胞来提高分化效率。这些TF的过表达将产生更同质和更有能力的干细胞群，其更适合于随后的肝分化。目标3将集中在肝细胞移植和提高肝细胞植入利用一个良好的描述归巢系统，是天然的肝脏。将对新鲜人肝细胞以及来自目的1和2的干细胞衍生的肝细胞进行修饰以过表达特异性归巢受体，并将其脾内注射到高度免疫缺陷小鼠中以研究肝移植和功能。这项研究将成为R 01资助申请的基础，该申请将研究从干细胞中开发临床有用的肝细胞的新方法。 公共卫生关系： 在美国，目前有近16，000人在等待肝脏移植，对器官的需求是供应的两倍多，每年有超过1，500人在等待名单上死亡（来自器官共享联合网络的统计数据，unos.org）。干细胞疗法治疗肝病是一种有前途的替代全器官移植，但仍需要进一步的研究，以提高安全性和有效性。