Mechanisms of macrophage death co-dependent on M. tuberculosis and IFN-a,b receptor

结核分枝杆菌和 IFN-a、b 受体共同依赖的巨噬细胞死亡机制

• 批准号: 10725738
• 负责人: CARL Francis NATHAN
• 金额: $ 25.43万
• 依托单位: WEILL MEDICAL COLL OF CORNELL UNIV
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10725738
• 关键词: Aconitic Acid; Animal Model; Antibiotics; Antimycobacterial Agents; Apoptosis; Binding; Biochemical; Biogenesis; COVID-19; CRISPR-mediated transcriptional activation; Carboxy-Lyases; Cathepsins; Cause of Death; Cell Death; Cell Death Induction; Cell Line; Cells; Cessation of life; Chemicals; Cods; Drug resistance; Enzymes; Future; G-Protein-Coupled Receptors; GPR84 gene; Genes; Genetic; Goals; Growth; IFNAR1 gene; Immune Response Genes; Impairment; In Vitro; Individual; Infection; Inflammation; Interferon Type I; Interferon alpha; Interferon-beta; Learning; Link; Lysosomes; Macrophage; Mediating; Medium chain fatty acid; Molecular; Monoclonal Antibodies; Mus; Mycobacterium tuberculosis; Necrotic Lesion; Orphan; Participant; Pathology; Pathway interactions; Peptide Hydrolases; Pharmaceutical Preparations; Pre-Clinical Model; Process; Proteins; Regimen; Resistance; Role; Signal Transduction; Speed; Testing; Tuberculosis; Virulent; Virus; bactericide; candidate identification; candidate validation; cytokine; drug-like compound; experimental study; gene product; genetic approach; improved; inhibitor; innovation; lysosomal Pro-X carboxypeptidase; overexpression; pathogen; receptor; response; success; transcription factor; tuberculosis treatment; type I interferon receptor

ABSTRACT Mycobacterium tuberculosis (Mtb) was the world’s single leading cause of death from infection before COVID- 19. Direct-acting antimycobacterial regimens are long, often toxic and plagued by emergence of drug resistance. Adjunctive therapies could potentially speed the cure of tuberculosis by targeting a process in the host that alters the host-pathogen interaction to the advantage of the host. One potential form of host-directed therapy would be to help macrophages better survive Mtb infection. In vitro, the death of Mtb-infected mouse macrophages is co- dependent on a type I interferon (IFN), IFN-beta, that macrophages produce in response to Mtb, along with an additional contribution by Mtb. This application seeks to identify specific molecular participants in Mtb-induced macrophage death that are downstream of the type I IFN receptor. We have identified potential molecular participants by two approaches—a CRISPR activation screen that restored Mtb-induced cell death to macrophages lacking the type I IFN receptor, and an innovative biochemical pulldown approach using a probe based on a small chemical compound that rescues Mtb-infected macrophages from Mtb-induced cell death without impairing the growth of Mtb. This application aims to validate the candidates genetically, test the ability of already existing inhibitors to recapitulate the effect of knocking them out, and then place the validated candidates on a mechanistic path. That would set the stage for future studies, beyond the scope of this application, to find, improve or develop drug-like inhibitors for tests in preclinical models of TB to see if they mitigate pathology and hasten cure when used in combination with direct-acting antimycobacterial agents.

摘要 结核分枝杆菌（Mtb）是COVID之前世界上唯一的主要感染死亡原因- 19.直接作用的抗分枝杆菌方案是长期的，通常是有毒的，并受到耐药性出现的困扰。 辅助疗法可能通过靶向宿主中改变肺结核的过程来加速肺结核的治愈。 宿主-病原体相互作用对宿主有利。宿主导向疗法的一种潜在形式是 帮助巨噬细胞更好地抵御结核杆菌感染。在体外，Mtb感染的小鼠巨噬细胞的死亡是协同的。 依赖于I型干扰素（IFN-β），巨噬细胞响应于Mtb而产生IFN-β，沿着 Mtb的额外捐款。本申请寻求鉴定Mtb诱导的免疫应答中的特定分子参与者。 I型IFN受体下游的巨噬细胞死亡。我们已经鉴定出潜在的分子 参与者通过两种方法-CRISPR激活筛选，恢复Mtb诱导的细胞死亡， 缺乏I型IFN受体的巨噬细胞，以及使用探针的创新生化下拉方法 基于一种小的化学化合物，可以拯救结核杆菌感染的巨噬细胞免于结核杆菌诱导的细胞死亡 而不损害Mtb的生长。这项申请旨在验证候选人的基因，测试能力， 已经存在的抑制剂，以概括敲除它们的效果，然后将经过验证的 这将为未来的研究奠定基础，超出本报告的范围。 应用，寻找，改进或开发药物样抑制剂，用于结核病临床前模型的测试，以观察它们是否 当与直接作用的抗分枝杆菌剂组合使用时，减轻病理学并加速治愈。