Role of Autophagy in Regulating Cytokine-Induced Macrophage Cell Death and Systemic Inflammatory Responses

自噬在调节细胞因子诱导的巨噬细胞死亡和全身炎症反应中的作用

基本信息

批准号：
10090558
负责人：
ANTHONY W ORVEDAHL
金额：
$ 16.72万
依托单位：
WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-02-03 至 2025-01-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10090558
关键词：
Aconitic Acid Address Anti-Inflammatory Agents Autophagocytosis Award Biochemical Biological Assay CRISPR screen Carboxy-Lyases Cell Death Cell Survival Cells Cessation of life Clinical Clustered Regularly Interspaced Short Palindromic Repeats Collaborations Communicable Diseases Complex Cytosol Degradation Pathway Disease Doctor of Philosophy Educational workshop Enabling Factors Ensure Environment Enzymes Event Exhibits Fibrinogen Future Gene Expression Profile Genes Goals Health Homeostasis Human Hypersensitivity Immune Immune response Immunity Immunology Immunoprecipitation Infection Inflammation Inflammatory Inflammatory Response Interferon Type II Interferons Interleukin-18 K-Series Research Career Programs Knock-out Lead Link Lysosomes Mediating Mediator of activation protein Medical Medical center Mentors Mentorship Microscopic Mitochondria Modality Modeling Molecular Molecular Genetics Morbidity - disease rate Multiple Organ Failure Mus Myelogenous Myeloid Cells Neonatology Outcome Pathology Pathway interactions Patients Pediatric Hospitals Pediatrics Physicians Physiological Play Production Protein Biochemistry Protein Kinase RIPK1 gene Regulation Research Research Personnel Resistance Resources Role Scientist Sepsis Shock Signal Transduction TNF gene Techniques Testing Therapeutic Tissues Training Training Activity Training Programs Tuberculosis Tumor Necrosis Factor Receptor Universities Washington Work analog antimicrobial antiviral immunity cell type cytokine genetic analysis genome-wide immune function improved improved outcome in vivo inflammatory disease of the intestine influenza infection live cell imaging macrophage medical schools mortality mouse model novel pathogen pediatric department prevent programs response skills systemic inflammatory response targeted treatment

项目摘要

PROJECT SUMMARY The goal of this Mentored Clinical Scientist Research Career Development Award (K08) is to provide a 5-year training pathway for Anthony Orvedahl, MD, PhD, to become an independent investigator. Dr. Orvedahl obtained the MD, PhD degree in the Medical Scientist Training Program at the UT Southwestern Medical Center, where he studied the role of autophagy in innate antiviral immunity. After training in Pediatrics and Infectious Diseases at St. Louis Children's Hospital/ Washington University, he joined the lab of Herbert “Skip” Virgin, MD, PhD, who is a renowned expert in antiviral immunity. Gary Silverman, MD, PhD, who will serve as the primary Co-Mentor, is a practicing physician-scientist in Neonatology with recognized expertise in mechanisms of cell death. In his preliminary work using genome-wide CRISPR screening, Dr. Orvedahl identified an important role for autophagy genes (including Atg5) in regulating IFNγ-induced cell death. A suppressor CRISPR screen on an Atg5-deficient background revealed the TNF pathway as a mediator of the hypersensitivity to cell death. However, TNF was insufficient on its own to trigger cell death, which indicated one or more additional IFNγ-induced factors contribute to TNF-induced death. Preliminary work suggests that one of these factors is the IFNγ-induced mitochondrial enzyme, immune regulated gene 1 (IRG1). Importantly, mice with myeloid cell-specific autophagy gene- deficiency exhibited markedly decreased survival in a TNF-induced model of shock. However, the specific type of cell death induced by IFNγ in combination with TNF, the mechanism of IRG1 and its enzymatic product itaconate, and the in vivo role of these factors during TNF-induced shock remain unclear. The proposed studies to address these questions in this K08 will require in-depth analyses of cell death and mitochondrial homeostasis using multiple microscopic, flow cytometric, and biochemical techniques. Washington University School of Medicine provides an ideal environment to pursue the training plan outlined in this proposal. The Department of Pediatrics has a longstanding commitment and track record of training independent physician-scientists. Dr. Orvedahl has established successful collaborations related to this project, and interacts frequently with investigators in the departments of Pediatrics and Pathology/ Immunology. An oversight committee has been formed that includes advisors with expertise in immunometabolism, autophagy in intestinal inflammation, protein biochemistry, and macrophage immune responses. The training plan incorporates technical workshops, as well as formal coursework on grantsmanship, which will facilitate transition to independence towards the end of the award period. The combination of mentorship, institutional resources, and focused training activities offer an unparalleled opportunity to achieve the goals of this proposal. The immediate research goals described herein are to: 1) investigate the mechanism of autophagy in protection against cytokine-induced macrophage cell death; 2) evaluate the role of a novel factor, IRG1, that links mitochondria to the mechanism of cell death; and 3) apply these findings in vivo to a model of fatal TNF-induced shock.

项目概要指导临床科学家研究职业发展奖 (K08) 的目标是提供为期 5 年的安东尼·奥维达尔 (Anthony Orvedahl) 博士成为一名独立研究者的培训途径。德克萨斯大学西南医学中心医学科学家培训项目的医学博士、博士学位，其中在接受儿科和传染病培训后，他研究了自噬在先天抗病毒免疫中的作用。在华盛顿大学圣路易斯儿童医院，他加入了 Herbert “Skip” Virgin 医学博士、博士的实验室，他抗病毒免疫领域的著名专家加里·西尔弗曼（Gary Silverman）医学博士、博士将担任主要联合导师，是新生儿学领域的执业医师科学家，在细胞死亡机制方面拥有公认的专业知识。 Orvedahl 博士利用全基因组 CRISPR 筛选进行初步工作，确定了自噬的重要作用 Atg5 缺陷基因（包括 Atg5）调节 IFNγ 诱导的细胞死亡的抑制 CRISPR 筛选。背景揭示了 TNF 通路是细胞死亡超敏反应的介质。其本身不足以引发细胞死亡，这表明一种或多种额外的 IFNγ 诱导因素有助于初步研究表明，这些因素之一是 IFNγ 诱导的线粒体。酶，免疫调节基因 1 (IRG1) 重要的是，具有骨髓细胞特异性自噬基因的小鼠。缺乏症在 TNF 诱导的休克模型中表现出显着降低的存活率。 IFNγ联合TNF诱导的细胞死亡、IRG1及其酶产物的作用机制衣康酸，以及这些因素在 TNF 诱导的休克期间的体内作用仍不清楚。为了解决本 K08 中的这些问题，需要对细胞死亡和线粒体稳态进行深入分析使用多种显微镜、流式细胞仪和生物化学技术。医学系为实施本提案中概述的培训计划提供了理想的环境。儿科在培训独立医师科学家方面有着长期的承诺和记录。 Orvedahl 已与该项目建立了成功的合作，并经常与儿科和病理学/免疫学部门的研究人员已成立一个监督委员会。其成员包括具有免疫代谢、肠道炎症自噬、蛋白质方面专业知识的顾问培训计划还包括生物化学和巨噬细胞免疫反应。作为关于赠款的正式课程，这将有助于在本世纪末过渡到独立指导、机构资源和重点培训活动的结合提供了一个奖励期。实现本提案中描述的近期研究目标的无与伦比的机会。目的是： 1) 研究自噬在防止细胞因子诱导的巨噬细胞死亡方面的机制； 2) 评估将线粒体与细胞死亡机制联系起来的新因子 IRG1 的作用；以及 3) 应用这些发现在体内建立了致命性 TNF 诱导休克模型。