MyD88-dependent B cell dysfunction and autoimmunity during chronic liver disease

项目摘要/摘要 NIDDK导师研究科学家职业发展奖(K01)的目标是提供资金 对于首席研究员(PI)Manoj Thapa博士，正在发展成为一项独立的生物医学研究 肝病和肝脏免疫学领域的科学家。在加入阿拉什·格拉库博士的实验室后 埃默里大学耶克斯国家灵长类研究中心，PI发起了一个新的项目，意在 在肝病期间识别导致T和B细胞分化的因素。在这份K01提案中，Pi将 以他以前的专业知识为基础，研究细胞免疫反应的动力学，重点是 慢性丙型肝炎相关慢性肝病中B细胞功能紊乱的机制 感染(丙型肝炎)、酒精性肝病(ALD)和非酒精性脂肪性肝炎(NASH)。PI将分配75% 他在这份K01申请中建议的研究时间和25%的时间用于教学培训 参与研讨会、实验室会议、手稿准备、赠款撰写以及国内和国际会议 会议。 本项目将重点研究MyD88特异性Toll样受体(TLR)激活和BAFF/BLyS的作用 B细胞介导的肝病和自身免疫性疾病发病机制的信号转导。末梢出血的发生率 在美国，阶段性肝病(ESLD)正在增加，并造成严重的经济和临床负担 因为预计它将在全球范围内影响超过3亿人。慢性丙型肝炎、ALD和NASH 占ESLD病例的大多数，慢性丙型肝炎与肝脏的发病率最高 移植。原位肝移植是治疗原发性系统性红斑狼疮的最佳方法，费用较高。 同时受益的相对较少。了解细胞免疫反应的机制 导致纤维化，而ESLD的进展对于有效的治疗干预至关重要。圆周率将 从两个不同的方向来处理这些问题。首先，通过使用小鼠实验性肝脏模型 纤维化，PI将描述MyD88特异性TLR信号和BAFF/BLyS信号在B细胞中的作用 与肝病相关的功能障碍、肝纤维化和自身免疫性疾病。然后，他将建立他的职业生涯 进入人类翻译科学，以提高对人类慢性疾病细胞免疫反应的理解 肝病包括丙型肝炎、酒精性肝病和非酒精性肝病。这些疾病虽然不同，但有共同的主题 (即纤维化/肝硬变)，因此在本研究中提出比较和对比病毒的影响 感染(丙型肝炎病毒)，一种非感染的炎症(NASH)，以及没有病毒和正在进行的 移植时的炎症性侮辱(ALD)(至少需要戒酒6个月 在埃默里移植中心移植之前)对B细胞激活和功能的影响。该应用程序将提供一个 靶向治疗逆转白血病进展及临床后遗症研究的免疫学基础 肝纤维化。 在K01奖项期间，Pi计划进一步掌握人类肝脏领域的技能和熟练程度 生物学、慢性肝病和肝脏免疫学，参加埃默里大学提供的教学课程 大学和其他公认的专业组织，负责任的研究行为培训， 参加翻译研究、初级教师发展课程和教师证书课程 发展系列讲座。PI计划在人体标本中获得深入的培训(肝脏移植和 人体组织的血液处理和免疫学分析，以了解复杂的相互作用 在多因素疾病中免疫系统和肝脏之间的关系，包括肝纤维化、肝硬化、ESLD和 自身免疫性疾病。此外，PI还计划积极参加每周或每月的系列研讨会 在埃默里大学提供，并参加关于纤维化、信号通路的年度全国会议， 免疫学、肝病和丙型肝炎病毒(丙型肝炎病毒)。此外，PI将利用多个 在埃默里大学举办赠款撰写研讨会，并计划通过R21机制申请NIH拨款 第三年为R01，第四年为R01，在第四年和第五年期间根据需要提交修订。聚酰亚胺 将从包括丙型肝炎病毒感染和肝脏专家阿拉什·格拉库伊博士在内的导师团队中受益匪浅 免疫学，马克斯·库珀博士，著名的B细胞免疫学家，弗兰克·阿纳尼亚博士， 肝脏病学和胃肠病学，以及著名免疫学家巴厘普伦德兰博士。这群杰出的 科学家将为PI提供所需的额外专业知识和培训，以实现他的短期目标 实现科学独立和成为老牌生物医学的长期目标 研究科学家并制定研究计划，以确定与慢性疾病有关的基本问题 肝脏疾病和相关的自身免疫性疾病。

PROJECT SUMMARY/ABSTRACT The goal of this NIDDK Mentored Research Scientist Career Development (K01) Award is to provide funding for the principal investigator (PI), Dr. Manoj Thapa, developing into an independent biomedical research scientist in the field of liver disease and liver immunology. After joining the Dr. Arash Grakoui’s laboratory at Yerkes National Primate Research Center of Emory University, the PI initiated a new project with the intention of identifying factors that lead to T and B cell differentiation during liver disease. In this K01 proposal, PI will build upon his previous expertise and study the dynamics of cellular immune responses focusing on the mechanisms of B cell dysregulations during chronic liver diseases associated with chronic hepatitis C virus (HCV) infection, alcohol liver disease (ALD) and nonalcoholic steatohepatitis (NASH). PI will be allocating 75% of his time for the research as proposed in this K01 application and 25% of the time to didactic trainings such as participation in seminars, lab meetings, manuscript preparation, grant writing, and national and international conferences. This project will focus on the contribution of MyD88-specific toll like receptor (TLR) activation and BAFF/BLyS signaling in the B cell-mediated pathogenesis of liver disease and autoimmune disorder. The incidence of end- stage liver disease (ESLD) is increasing in the United States and poses a serious economic and clinical burden as it is expected to afflict more than 300 million individuals worldwide. Chronic HCV infection, ALD, and NASH account for the majority of ESLD cases, with chronic HCV associated with the highest incidence of liver transplantation. The orthotopic liver transplantation is the optimal treatment for ESLD and carries a high cost while benefitting relatively few. Understanding the mechanisms of the cellular immune responses that contributes to fibrosis and the progression of ESLD is critical for effective therapeutic intervention. PI will approach these questions from two different directions. First, by using murine models of experimental liver fibrosis, PI will delineate the role of MyD88-specific TLR signaling and BAFF/BLyS signaling in B cell dysfunction, liver fibrosis and autoimmune disorder associated with liver disease. Then, he will build his career into human translational science to improve an understanding of cellular immune response in human chronic liver diseases including HCV, ALD and NASH patients. These diseases while different have common themes (i.e. fibrosis/cirrhosis) and therefore proposed in this study to compare and contrast the effect of a viral infection (HCV), an inflammation without infection (NASH) and the absence of virus and an on-going inflammatory insult at the time of transplant (ALD) (alcohol abstinence is required for a minimum of 6 months prior to transplant at Emory Transplant Center) on B cell activation and function. The application will provide an immunological basis for the study of targeting therapeutics to reverse the progression and clinical sequelae of liver fibrosis. During the K01 award, PI plans to further master skills and acquire proficiency in the area of human liver biology, chronic liver disease and liver immunology by participating in didactic courses offered by the Emory University and other recognized professional organizations, training in responsible conduct of research, enrolling in Certificate Program in Translational Research, Junior Faculty Development Course and Faculty Development Lecture Series. PI plans to acquire in-depth trainings in human specimens (liver explants and blood) processing and immunological analysis of human tissues in understanding of the complex interaction between the immune system and the liver in multifactorial disease including liver fibrosis, cirrhosis, ESLD, and autoimmune disorders. In addition, PI also plans to actively participate in weekly or monthly seminar series available at Emory University and attend annual national conferences on Fibrosis, Signaling pathways, Immunology, Liver Diseases, and Hepatitis C Virus (HCV). Furthermore, PI will take advantage of multiple grant-writing workshops available at Emory University and plan to apply for NIH grants via the R21 mechanism in the third year, and R01 in the fourth year submitting revisions as needed during the fourth and fifth years. PI will greatly benefit from a team of mentors including Dr. Arash Grakoui, an expert in HCV infection and liver immunology, Dr. Max Cooper, a foremost B cell immunologist, Dr. Frank Anania, an expert clinician in hepatology and gastroenterology, and Dr. Bali Pulendran, an established immunologist. This group of eminent scientists will provide additional expertise and training needed for the PI to achieve his short-term goal of achieving the scientific independence as well as the long-term goal of becoming an established biomedical research scientist and develop a research program to determine the fundamental questions regarding chronic liver disease and associated autoimmune disorders.