STAT3-dependent manipulation of host transcription and immune responses by Salmonella

沙门氏菌对宿主转录和免疫反应的 STAT3 依赖性操纵

基本信息

批准号：
9760288
负责人：
Kyle Dane Gibbs
金额：
$ 3.68万
依托单位：
DUKE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-02-01 至 2022-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9760288
关键词：
Amino Acids Anti-Bacterial Agents Autoimmune Diseases Autoimmunity Automobile Driving B-Lymphocytes Bacteria Binding Biochemistry Biological Assay Cell Differentiation process Cell Line Cell physiology Cells Cessation of life Communicable Diseases Complex Cytokine Receptors Data Exhibits Fluorescence Polarization Future Gastroenteritis Genes Genetic Transcription Genetic Variation Genotype Goals Growth Factor Hour Human Human Genetics Hypoxia IL6ST gene Immune response Immunity Infection Interleukin-10 Investigation Lead Measures Metabolic Modeling Mus Mutate Mutation Pathway interactions Phosphorylation Phosphotransferases Physiological Processes Process Process Measure Proteins Proteomics Regulation Research STAT3 gene Salmonella Salmonella enterica Salmonella infections Salmonella typhimurium Signal Pathway Signal Transduction Signal Transduction Pathway Stat3 protein Structure Testing Transfection Tyrosine Phosphorylation Vacuole Virulence base cofactor cytokine enzyme activity genetic variant genome wide association study improved in vivo infected B cell knock-down member non-typhoidal Salmonella novel pathogenic bacteria receptor response transcription factor transcriptome sequencing transcriptomics tumorigenesis

项目摘要

Project Summary/Abstract Background: Across the globe, non-typhoidal serovars of the intracellular bacterial pathogen Salmonella enterica caused 153 million cases of gastroenteritis and 57,000 deaths in 2010. S. enterica enterica ser. Typhimurium, the most prevalent non-typhoidal Salmonella (NTS) serovar, creates a durable niche inside host cell vacuoles by injecting effector proteins that subvert host signal transduction pathways. Recently, S. Typhimurium has been shown to promote its intracellular survival and replication by activating the host transcription factor STAT3. While STAT3 is known to promote cell differentiation, survival, and proliferation in response to cytokines and growth factors, it is unknown how Salmonella drives STAT3 phosphorylation or how STAT3 renders host cells more permissive to S. Typhimurium. Goal/Preliminary Data: I aim to understand the mechanism and function of STAT3 phosphorylation during Salmonella infection. Recently, our lab discovered that an S. Typhimurium effector, SarA, physically associates with STAT3 and induces STAT3 phosphorylation. However, SarA lacks identifiable structural homology to any known kinase domain, so I hypothesize that SarA initiates a signaling pathway to phosphorylate STAT3. Further, S. Typhimurium ∆sarA exhibits reduced intracellular replication in human cells and infected mice. This SarA-dependent reduction is recapitulated by knockdown or inhibition of host STAT3 during wild-type infection, indicating that SarA promotes virulence by activating STAT3. Hence, I hypothesize that STAT3- targets promote bacterial replication in the Salmonella-containing vacuole (SCV). Approach: I will determine how SarA-interacting partners, identified by proteomics, interact with phosphorylated SarA motifs to drive STAT3 tyrosine phosphorylation. In parallel, I will verify potential SarA to STAT3 signaling components or regulators that were identified through common human genetic variants associated with the level of STAT3-target activation. Finally, I will determine how activated STAT3 promotes S. Typhimurium intracellular replication, by ascertaining how SarA-dependent transcriptional changes, identified by transcriptomics analysis, promote intracellular bacterial replication. Significance: Uncovering how S. Typhimurium activates STAT3 to manipulate host transcription and improve its intracellular niche will enhance our understanding of bacteria-host interaction and the function of the STAT3 pathway, a crucial player in autoimmunity and oncogenesis.

项目概要/摘要背景：在全球范围内，细胞内细菌病原体的非伤寒血清型 2010 年，肠道沙门氏菌导致 1.53 亿例胃肠炎病例和 57,000 人死亡。伤寒肠杆菌，最流行的非伤寒沙门氏菌 (NTS) 血清型，通过注入颠覆宿主的效应蛋白，在宿主细胞液泡内创建一个持久的生态位最近，鼠伤寒沙门氏菌已被证明可以促进其信号转导途径。通过激活宿主转录因子 STAT3 来实现细胞内的存活和复制。 STAT3 已知可响应细胞因子促进细胞分化、存活和增殖和生长因子，尚不清楚沙门氏菌如何驱动 STAT3 磷酸化或 STAT3 如何驱动使宿主细胞更容易感染鼠伤寒沙门氏菌。目标/初步数据：我的目标是了解STAT3的机制和功能最近，我们的实验室发现了沙门氏菌感染期间的磷酸化。效应器 SarA 与 STAT3 物理结合并诱导 STAT3 磷酸化。 SarA 与任何已知的激酶结构域都缺乏可识别的结构同源性，所以我很挣扎 SarA 启动磷酸化 STAT3 的信号通路，此外，鼠伤寒沙门氏菌 ΔsarA。在人类细胞和受感染的小鼠中表现出细胞内复制减少。通过在野生型感染期间敲低或抑制宿主 STAT3 来重现减少，表明 SarA 通过激活 STAT3 来增强毒力，因此，我追踪了 STAT3-。目标促进含有沙门氏菌的真空 (SCV) 中的细菌复制。方法：我将确定通过蛋白质组学鉴定的 SarA 相互作用伙伴如何与磷酸化 SarA 基序驱动 STAT3 酪氨酸磷酸化同时，我将验证。通过共同鉴定的潜在 SarA 至 STAT3 信号传导成分或调节器人类遗传变异与 STAT3 靶标激活水平相关。确定激活的 STAT3 如何促进鼠伤寒沙门氏菌细胞内复制，通过通过转录组学分析确定 SarA 依赖性转录变化是如何发生的，促进细胞内细菌的复制。意义：揭示鼠伤寒沙门氏菌如何激活 STAT3 来操纵宿主转录并改善其细胞内生态位将增强我们对细菌与宿主相互作用的理解 STAT3 通路的功能，在自身免疫和肿瘤发生中发挥着重要作用。