The Role of CD8+ T Cells In The Development of Nonalcoholic Fatty Liver Disease

描述(申请人提供)：我的职业目标是在学术界领导一个成功的实验室，结合细胞、动物和临床研究，解决先天性免疫系统对肥胖和相关代谢紊乱的影响，以及营养对免疫细胞功能的影响。我正在申请导师职业发展奖，因为它提供了一个转变为独立调查员的绝佳机会。目前，我是Alyssa Hats博士实验室的博士后实习生，在那里我能够提高我的分子技术，并获得处理免疫细胞和肥胖症和高脂血症动物模型的经验。我已经能够建立自己的独立研究项目，旨在研究肝脏CD8+T细胞在非酒精性脂肪性肝病发展中的作用。在目前的提议下，将获得一些研究技能，包括RNA测序、抗体耗竭、过继细胞转移研究和流式细胞术。同样在哈斯蒂博士的指导下，我将继续接受科学方面的培训 沟通和专业技能，成为一名独立调查员所必需的。范德比尔特大学医学中心的科学环境非常适合帮助我实现研究免疫系统和代谢性疾病的长期职业目标。有大量的共享资源和核心设施(Vantage核心、流式细胞仪核心)和大量的协作机会可用。我之前的培训机会和科学技能为我提供了在肥胖、糖尿病、心血管疾病和非酒精性脂肪性肝病方面的坚实背景，这些都是执行我提议的研究所必需的。因此，在这次培训中获得的知识和科学技能将有助于我为未来成为学术界独立研究科学家的目标做好准备。摘要随着代谢性疾病的大量增加，非酒精性脂肪性肝病(NAFLD)在当今社会越来越常见。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)以肝脏脂肪变性和混合性炎细胞浸润为特征，是肝纤维化进展的主要预测指标。最近，纳什已经 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Kupffer细胞在NAFLD中的作用受到了很大的关注，但对淋巴细胞亚群及其在NAFLD中的作用的关注却很少。在肥胖条件下，CD8+T细胞可以调节脂肪组织中巨噬细胞的渗透。我们的初步数据首次证明高脂血症可诱导表达独特的Th2和细胞毒表型的肝脏CD8+T细胞的早期渗透。这种肝淋巴细胞的浸润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，本项目的具体目标是：1)确定从脂肪变性向NASH转变过程中肝脏CD8+T细胞群的表型；2)在体内确定CD8+T细胞对NASH发生发展的影响程度；3)在体内确定IL-10对从肝脏脂肪变性向NASH转变的影响程度。我们的假设是：1)早期渗透的CD8+T细胞表达一种细胞毒和Th2表型，驱动巨噬细胞的募集和促纤维化表型；2)CD8+T细胞的耗竭将减少巨噬细胞的募集和促纤维化表型；而CD8+T细胞的加入将增加肝脂肪变性条件下巨噬细胞的促纤维化表型；3)在NAFLD条件下，IL-10的缺失将减少肝巨噬细胞的促纤维化表型。从这项提案中获得的数据有望为NASH预防和治疗的潜在治疗目标的开发提供重要的洞察力，并降低与这些疾病相关的医疗成本。(摘要结束)

DESCRIPTION (provided by applicant): 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. (End of Abstract)