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

• 批准号: 9892285
• 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
• 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/9892285
• 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; Failure; Fibrinogen; Future; Genes; Genetic Transcription; Goals; Health; Homeostasis; Human; Hypersensitivity; Immune; Immune response; Immunity; Immunology; Immunoprecipitation; Infection; Inflammation; Inflammatory; Inflammatory Response; Influenza; 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; 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; live cell imaging; macrophage; medical schools; mortality; mouse model; novel; pathogen; pediatric department; prevent; programs; response; skills; systemic inflammatory response; targeted treatment

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.

项目摘要 这个受过指导的临床科学家研究职业发展奖（K08）的目标是提供5年 训练Anthony Orvedahl，医学博士，博士，成为一名独立研究员。 Orvedahl博士获得了 UT西南医学中心医学科学家培训计划的医学博士学位，博士学位 他研究了自噬在先天抗病毒免疫学中的作用。经过儿科和传染病训练 在圣路易斯儿童医院/华盛顿大学，他加入了赫伯特“跳过”维尔京的实验室，博士，博士 是抗病毒免疫的著名专家。医学博士的加里·西尔弗曼（Gary Silverman）将担任主要院长 是新生儿学的实用身体科学家，在细胞死亡机制方面具有公认的专业知识。在他的 使用全基因组CRISPR筛选的初步工作，Orvedahl博士确定了自噬的重要作用 调节IFNγ诱导的细胞死亡的基因（包括ATG5）。 ATG5缺陷的抑制器CRISPR屏幕 背景揭示了TNF途径是对细胞死亡超敏反应的介体。但是，TNF是 本身不足以触发细胞死亡，这表明一个或多个IFNγ诱导的因子有助于 致TNF诱发的死亡。初步工作表明，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，具有髓样细胞特异性自噬基因的小鼠 - 在TNF诱导的冲击模型中，缺乏显着降低了存活率。但是，特定类型 IFNγ与TNF联合诱导的细胞死亡，IRG1及其酶产物的机理 Itaconate，以及这些因素在TNF诱导的冲击过程中的体内作用尚不清楚。提出的研究 在此K08中解决这些问题将需要对细胞死亡和线粒体稳态进行深入分析 使用多个显微镜，流式细胞仪和生化技术。华盛顿大学学校 医学提供了一个理想的环境，可以追求本提案中概述的培训计划。部门 儿科在培训独立的身体科学家方面有着长期的承诺和记录。博士 Orvedahl已经建立了与该项目相关的成功合作，并经常与 儿科和病理/免疫学部门的研究人员。监督委员会已经 成立的，包括具有免疫代谢专业知识，肠道注射自噬的顾问，蛋白质 生物化学和巨噬细胞免疫反应。培训计划还包括技术研讨会 作为授予技巧的正式课程，这将有助于向独立过渡到结束 奖励期。心态，机构资源和集中培训活动的结合提供了 无与伦比的机会实现该提案的目标。本文描述的直接研究目标 为：1）研究自噬在保护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理联系起来的新型因素IRG1的作用； 3）申请 这些发现在体内与致命TNF诱导的休克模型的模型。