The role of CD8+ T cells in the development of Nonalcoholic Fatty Liver Disease

CD8 T 细胞在非酒精性脂肪肝发展中的作用

• 批准号: 8616468
• 负责人: Arion Kennedy
• 金额: $ 11.3万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-01-15 至 2018-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8616468
• 关键词: Academia; Academic Medical Centers; Address; Adipose tissue; Adoptive Cell Transfers; Animal Model; Animals; Antibodies; Attention; Award; Benign; CD8B1 gene; Carcinoma; Cardiovascular Diseases; Cell physiology; Cells; Characteristics; Chronic; Cirrhosis; Clinical Research; Communication; Core Facility; Coronary; Country; Cytoplasm; Data; Deposition; Development; Diabetes Mellitus; Disease; Environment; Fatty Liver; Fibrosis; Flow Cytometry; Future; Gene Expression; Goals; Grant; Health Care Costs; Hepatic; Hepatocyte; Histopathology; Hyperlipidemia; Immune; Immune system; Incidence; Infiltration; Inflammation; Inflammatory; Insulin Resistance; Interleukin-10; K-Series Research Career Programs; Knowledge; Kupffer Cells; Laboratories; Lead; Liver; Liver Fibrosis; Liver diseases; Lymphocyte; Lymphocyte Subset; Mentors; Mentorship; Metabolic Diseases; Modeling; Molecular; Mus; Non obese; Non-Insulin-Dependent Diabetes Mellitus; Nutrient; Obesity; Phenotype; Population; Postdoctoral Fellow; Prevalence; Prevention; RNA Sequences; Research; Research Personnel; Research Project Grants; Resource Sharing; Risk Factors; Role; Scientist; Societies; Solid; T-Lymphocyte; Techniques; Testing; Tissue Inhibitor of Metalloproteinase-1; Training; Triglycerides; Work; Wound Healing; abstracting; base; career; cytotoxic; disorder prevention; effective therapy; experience; in vivo; insight; liver inflammation; liver injury; macrophage; monocyte; mouse model; non-alcoholic fatty liver; nonalcoholic steatohepatitis; prevent; response; skills; therapeutic target

PROJECT SUMMARY/ABSTRACT Candidate My career goal is to direct a successful laboratory in academia, incorporating cellular, animal, and clinical research to address the impact of the innate immune system on obesity and associated metabolic disorders and the impact of nutrients have on immune cell function. I am applying for the Mentored Career Development Award because it offers an excellent opportunity to transition into an independent investigator. Currently I am a post-doctoral trainee in Dr. Alyssa Hasty's laboratory where I have been able to enhance my molecular techniques and gain experience working with immune cells and animal models of obesity and hyperlipidemia. I have been able to establish my own independent research project aimed at the role of hepatic CD8+ T cells in the development of non-alcoholic fatty liver disease. A number of research skills will be obtained under the present proposal, including RNA sequencing, antibody depletion, adoptive cell transfer studies, and flow cytometry. Also under the mentorship of Dr. Hasty, I will continue to obtain training in scientific communication and professional skills, necessary to become an independent investigator. The scientific environment of Vanderbilt University Medical Center is ideally suited to help me achieve my long career goal to study the immune system and metabolic disease. There are numerous shared resources and core facilities (VANTAGE core, Flow cytometry core) and a vast number of collaborative opportunities available. My previous training opportunities and scientific skills have provided me with a solid background in obesity, diabetes, cardiovascular disease, and nonalcoholic fatty liver disease, necessary to execute my proposed studies. Thus, the knowledge and scientific skills obtained during this training award will help prepare me for my future goal of becoming an independent research scientist in academia. Abstract With the vast increase of metabolic disease, nonalcoholic fatty liver disease (NAFLD) is progressively more common in today's society. NAFLD ranges from hepatic steatosis to cirrhosis. Nonalcoholic steatohepatitis (NASH) is characterized by hepatic steatosis and mixed inflammatory cell infiltration and is a major predictor of fibrotic progression. Recently, NASH has been identified as a risk factor for cardiovascular disease. It is well known that immune cells are key contributors to inflammation and fibrosis. While much attention has been placed on the role of kupffer cells in NAFLD, limited attention has been placed on the subpopulations of lymphocytes and their contribution to NAFLD. Under obese conditions, CD8+ T cells have been found to regulate macrophage infiltration in adipose tissue. Our preliminary data are the first to demonstrate that hyperlipidemia induces an early infiltration of hepatic CD8+ T cells that express a unique Th2 and cytotoxic phenotype. This hepatic lymphocyte infiltration correlates with an increase in hepatic inflammatory monocytes and fibrogenic markers. However, the distinct role of these immune cells in NASH is unclear. Thus, the Specific Aims of this project are: 1) To identify the phenotype of hepatic CD8+ T-cell populations during the transition from steatosis to NASH; 2) To determine in vivo the extent to which CD8+ T-cells impact the development of NASH; 3)To determine in vivo the extent to which IL-10 impacts the transition from hepatic steatosis to NASH. Our hypotheses are 1) early infiltrating CD8+ T-cells express a cytotoxic and Th2 phenotype which drives the recruitment and the profibrogenic phenotype in macrophages; 2) depletion of CD8+ T cells will reduce the recruitment and profibrogenic phenotype of macrophages; while addition of CD8+ T cells will increase the profibrogenic phenotype of macrophages under conditions of hepatic steatosis; 3) depletion of IL-10 will decrease the profibrogenic phenotype in hepatic macrophages under NAFLD conditions. Data obtained from this proposal are expected to provide critical insight for the potential development of therapeutic targets for the prevention and treatment of NASH, and reduce health care costs related to these disorders.

项目摘要/摘要 侯选人 我的职业目标是在学术界领导一个成功的实验室，将细胞， 动物和临床研究，以解决先天免疫系统对肥胖和 相关的代谢紊乱和营养物质对免疫细胞功能的影响。我是 申请导师职业发展奖，因为它提供了一个极好的机会 转变为一名独立调查员。目前我是博士后实习生，正在攻读博士后。 阿丽莎·哈斯蒂的实验室，在那里我能够提高我的分子技术和 获得处理免疫细胞和肥胖症和高脂血症动物模型的经验。我 我已经能够建立自己的独立研究项目，旨在发挥肝脏的作用 CD8+T细胞在非酒精性脂肪性肝病发病中的作用一些研究技能 将根据本提议获得，包括RNA测序、抗体耗竭、 过继细胞转移研究和流式细胞术。同样在哈提斯博士的指导下，我会 继续接受科学交流和专业技能方面的培训，这是 成为一名独立调查员。范德比尔特大学的科学环境 医疗中心非常适合帮助我实现研究免疫的长期职业目标 系统和代谢性疾病。有许多共享的资源和核心设施 (Vantage核心、流式细胞仪核心)和大量的协作机会 可用。我以前的培训机会和科学技能为我提供了坚实的 肥胖、糖尿病、心血管疾病和非酒精性脂肪肝的背景， 来完成我所提议的研究。因此，所获得的知识和科学技能 这个训练奖将帮助我为未来成为一名独立的人的目标做好准备 学术界的研究科学家。 摘要 随着代谢性疾病的大量增加，非酒精性脂肪性肝病(NAFLD) 在今天的社会中越来越普遍。非酒精性脂肪肝的范围从肝脏脂肪变性到 肝硬变。非酒精性脂肪性肝炎(NASH)的特点是肝脏脂肪变性和混合性 炎性细胞浸润，是纤维化进展的主要预测指标。最近，纳什 已被确定为心血管疾病的危险因素。众所周知， 免疫细胞是炎症和纤维化的关键因素。虽然很多人都在关注 关于Kupffer细胞在NAFLD中的作用，人们对Kupffer细胞的关注有限 淋巴细胞亚群及其在NAFLD中的作用。在肥胖条件下，CD8+ 已发现T细胞可调节脂肪组织中巨噬细胞的渗透。我们的预赛 首次有数据表明高脂血症会导致肝脏CD8+的早期渗透。 表达独特的Th2和细胞毒表型的T细胞。肝淋巴细胞的浸润性 与肝脏炎性单核细胞和纤维化标志物的增加有关。 然而，这些免疫细胞在NASH中的不同作用尚不清楚。因此，具体的目标是 本项目的主要内容是：1)确定肝组织CD8+T细胞亚群的表型 从脂肪变性到NASH的转变；2)在体内确定CD8+T细胞的程度 影响NASH的发展；3)在体内确定IL-10对NASH的影响程度 从肝脏脂肪变性过渡到NASH。我们的假设是1)早期渗入CD8+T细胞 表达一种细胞毒和Th2表型，驱动募集和促纤维化 巨噬细胞的表型；2)CD8+T细胞的耗尽会减少募集和 巨噬细胞的促纤维化表型；而CD8+T细胞的加入会增加 肝脂肪变性条件下巨噬细胞促纤维化表型；3)耗竭 IL-10对NAFLD大鼠肝巨噬细胞促纤维化表型的影响 条件。从这项提案中获得的数据预计将为 预防和治疗NASH的治疗靶点的潜在开发，以及 降低与这些疾病相关的医疗费用。