Molecular Pathways of Programmed Cell Death And Viral Cytopathicity

程序性细胞死亡和病毒细胞病变的分子途径

• 批准号: 10697667
• 负责人: michael j lenardo
• 金额: $ 63.1万
• 依托单位: NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10697667
• 关键词: Allergic; Antigens; Apoptosis; Autoimmune; Autoimmune Diseases; Autoimmunity; Biological; Biological Process; CRISPR/Cas technology; Candidate Disease Gene; Cell Death; Cell Survival; Cells; Ceramides; Cessation of life; Coin; Complex; Contracts; Coupled; Disease; Embryonic Development; Equation; Equilibrium; Event; Exposure to; Genes; Gold; Graft Rejection; Homeostasis; Human; Immune response; Immune system; Immunity; Immunosuppressive Agents; In Vitro; Knock-out; Lead; Libraries; Life; Lipids; Liquid Chromatography; Lymphocyte; Lymphoproliferative Disorders; Malignant lymphoid neoplasm; Maps; Mass Spectrum Analysis; Medicine; Methods; Modification; Molecular; Mus; Outcome; Pathway interactions; Phase; Play; Process; Proteins; Proteome; Reagent; Receptor Signaling; Research; Resolution; Resources; Rest; Role; Sampling; Signal Transduction; Specificity; Spectrometry; Speed; System; T-Cell Proliferation; T-Cell Receptor; T-Lymphocyte; Therapeutic Intervention; Transcriptional Activation; Viral; base; experimental study; genome-wide; interest; ion mobility; lipidomics; liquid chromatography mass spectrometry; mass spectrometer; metabolome; metabolomics; pathogen; programs; receptor; screening; side effect; therapeutic development; therapeutically effective

Programed cell death (PCD) is an indispensable process that takes place through a variety of different mechanisms in multicellular life. Apoptosis and necroptosis, which are essential for embryonic development and many important biological processes that rely on continuous cellular turn-over, are recognized as the two major forms of PCD that contribute to the contraction phase of immune responses. In the periphery, stimulation of the T cell antigen receptor (TCR) can have several different outcomes, including T cell survival, activation, and proliferation, or PCD. However, T cell proliferation and contraction during an immune response must be balanced to allow for both protection against pathogens and avoidance of autoimmunity and the excessive accumulation of cells that might predispose to lymphoid cancer. Although primary stimulation of TCR in resting T cells and restimulation of TCR in activated cells share many early signaling events, there are significant differences that result in either life or death respectively. We coined the term TCR restimulation-induced cell death (RICD) to conceptually distinguish the TCR process resulting in apoptosis from that leading to activation and proliferation. Clonal specificity is the important feature of RICD, which makes it an appealing method for tolerance induction in autoimmune diseases. Compared with conventional immunosuppressant medicines that debilitate the whole immune system and have many side effects, the RICD-based tolerance strategy targets only undesired T cells while keeping the general immune system intact. Thus, tolerogenic induction of antigen- specific RICD may be an effective therapeutic intervention for progressive autoimmune disorders. It is well known that the T cell undergoes dramatic changes immediately after a resting or naive cell is exposed to an antigen. During the TCR signaling cascade, the related proteins are activated, along with transcriptional activation of several T cell-specific genes, causing a wide scale modification of its metabolome. Currently, few details are known about metabolomics during this process. Liquid chromatography (LC) coupled with mass spectrometry (MS) has become the gold standard in various omics fields. However, the coverage of proteomes or metabolomes in complex biological samples remains challenging due to limited speed, sensitivity, and resolution of current mass spectrometers. Adding trapped ion mobility spectrometry (TIMS) to the equation unlocks the parallel accumulation serial fragmentation acquisition method to provide extremely high MS/MS speed and sensitivity, requiring minimal sample amounts. We have acquired a start-of-the-art TimsTOF LC-MS spectrometer system for lipidomics and metabolomics research. In our first project, we are studying the role lipids play in cell death or survival decisions when T cells are sensitive to RICD. Our preliminary in vitro findings show differences in lipid ceramide levels in T cells undergoing activation versus RICD. Further studies will help to map lipid pathways that are involved with the TCR signaling complex. Additionally, we are carrying out genome-wide CRISPR-cas 9 knockout library screening to identify candidate genes encoding proteins that promote cell survival under selected death-inducing circumstances, such as death-receptor and RICD. Due to availability of resources and reagents, these experiments are being conducted using primary mouse cells, but we will confirm the experiments with human T cells once our genes of interest are identified. Blocking key parts of these pathways highly associated with autoimmune diseases could aid in therapeutic development.

程序性细胞死亡（PCD）是多细胞生命中一个不可缺少的过程，它通过多种不同的机制发生。细胞凋亡和坏死下垂是胚胎发育和许多依赖于连续细胞翻转的重要生物过程所必需的，被认为是两种主要的PCD形式，有助于免疫反应的收缩阶段。在外周，T细胞抗原受体（TCR）的刺激可产生几种不同的结果，包括T细胞存活、活化和增殖（PCD）。然而，在免疫反应过程中，T细胞的增殖和收缩必须得到平衡，以防止病原体和避免自身免疫以及可能易患淋巴样癌的细胞的过度积累。虽然静息T细胞中TCR的初次刺激和活化细胞中TCR的再刺激有许多共同的早期信号事件，但分别导致生存或死亡的显著差异。我们创造了术语TCR再刺激诱导细胞死亡（RICD），从概念上区分导致凋亡的TCR过程和导致活化和增殖的TCR过程。克隆特异性是RICD的重要特征，这使其成为自身免疫性疾病耐受诱导的一种有吸引力的方法。传统的免疫抑制药物会削弱整个免疫系统并有许多副作用，与之相比，基于ricd的耐受策略只针对不需要的T细胞，同时保持一般免疫系统的完整。因此，抗原特异性RICD的耐受性诱导可能是进行性自身免疫性疾病的有效治疗干预措施。