Sulfide metabolism at the host microbiome interface

宿主微生物组界面的硫化物代谢

• 批准号: 10151704
• 负责人: David A Hanna
• 金额: $ 6.6万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-01 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10151704
• 关键词: Address; Affect; Age; Aging; Air; Animal Model; Antioxidants; Assimilations; Atmosphere; Behavior; Biogenesis; Caenorhabditis elegans; Cardiovascular Physiology; Cell Respiration; Cells; Colon; Colon Carcinoma; Complement; Cysteine; Data; Development; Diet; Dose; Electron Transport; Electrons; Energy Metabolism; Enzymes; Epigenetic Process; Epithelial Cells; Escherichia coli; Exposure to; Extravasation; Foundations; Genes; Genetic Transcription; Germ-Free; Glutathione; Glutathione Disulfide; Histone Acetylation; Histone Deacetylase; Homeostasis; Human; Hydrogen Sulfide; Hypoxia; Infection; Inflammation; Intestines; Knowledge; Laboratories; Larva; Life; Link; Longevity; Lysine; Maintenance; Mammalian Cell; Mediating; Metabolic Pathway; Metabolism; Methylation; Michigan; Microbe; Mitochondria; Modification; Molecular; Mus; Mutation; Neurons; Organism; Orthologous Gene; Oxidation-Reduction; Oxides; Pathway interactions; Physiological; Physiological Processes; Poison; Post-Translational Protein Processing; Process; Production; Proteins; Proteomics; RNA Interference; Reporting; Resistance; Respiration; Signal Transduction; Signaling Molecule; Stress; Sulfate; Sulfhydryl Compounds; Sulfides; Sulfites; Sulfur; Sulfur Metabolism Pathway; Testing; Toxic effect; Translations; Vasodilation; Water; base; biological adaptation to stress; cardioprotection; cell type; cytotoxic; dietary; dietary manipulation; electron energy; enzyme pathway; gastrointestinal function; genetic manipulation; gut bacteria; gut microbes; gut microbiome; histone methylation; histone modification; host microbiome; knock-down; metabolomics; microbial; microbiota; neuroregulation; oxidation; persulfides; respiratory; response; sensor; sulfate reducing bacteria; transcriptome sequencing; transcriptomics

Project Summary Hydrogen sulfide (H2S) is a redox-active signaling molecule that modulates electron transport and energy metabolism, and by largely unknown mechanisms, mediates neuro- and cardioprotection, vasodilation, the hypoxic response, protein translation, epigenetics, aging, and both patho- and physiological responses in colon. As H2S is also a respiratory poison at high doses, cells actively oxidize it to produce persulfides, sulfite, thiosulfate, (collectively referred to as reactive sulfur species), and sulfate. Oxidative modification of protein cysteine thiols to persulfides is postulated to be a primary mechanism by which H2S signals. The potential involvement of other reactive sulfur species in signaling is however, unknown. Studies in our laboratory have demonstrated that H2S impacts cellular redox metabolism via its dual effects on mitochondrial energetics, i.e. increasing electron flux at low, and inhibiting it at high concentrations. The highest exposure to exogenous H2S occurs in colon (0.2–2.4 mM) and is derived from gut resident microbiota, which synthesize sulfide. Preliminary data in our laboratory reveal that colonic epithelial cells quantitatively oxidize exogenous H2S to thiosulfate, and that the localization and expression levels of sulfide oxidation enzymes in murine colonocytes are strongly influenced by the presence or absence of sulfate reducing bacteria in the gut. I hypothesize that the dynamic interplay between host and microbial sulfur metabolites influences host metabolism and impacts longevity. Using Caenorhabditis elegans as a model organism, which has the full complement of orthologous genes involved in H2S biogenesis and oxidation found in humans and is readily amenable to genetic and dietary manipulation and to life-span analyses, I will test my hypothesis by addressing the following two aims. (i) I will characterize how alterations in H2S levels and reactive sulfur species impact organismal redox and histone modifications that are linked to longevity and stress response in C. elegans. H2S levels and its oxidative byproducts will be modulated by exogenous H2S, RNA interference knockdowns of host sulfide oxidation enzymes, and by diet, using Escherichia coli with deletions in specific genes involved in sulfur metabolism. I will assess how exogenous versus dietary H2S modulation affects organismal redox using the genetically encoded reduction-oxidation sensitive GRX1-roGFP2 sensor and track changes in histone methylation and acetylation status with reported links to redox, H2S availability, diet, and aging. (ii) I will investigate how differences in exogenous H2S exposure from air versus gut microbes affects the expression and localization of host sulfide oxidation enzymes, the abundance and type of reactive sulfur species, and their effect on lifespan. I will complement these studies with metabolomic and transcriptomic analyses to identify pathways that are impacted by H2S exposure. The successful completion of these studies will be foundational to our understanding of the interplay of sulfur metabolism at the host-microbe interface.

项目摘要 硫化氢（H2S）是一种氧化还原活性的信号分子，可调节电子传输和能量 新陈代谢，并且通过未知的机制介导神经保护，血管舒张， 结肠中的低氧反应，蛋白质翻译，表观遗传学，衰老以及病原体和身体反应。 由于H2S也是高剂量的呼吸毒物，因此细胞会积极地产生氧化硫酸盐，亚硫酸盐，硫酸盐， 硫代硫酸盐（统称为反应性硫种）和硫酸盐。蛋白质的氧化修饰 半硫化硫醇的半硫化物被张贴为H2S信号的主要机制。潜力 然而，其他反应性硫物种参与信号传导是未知的。在我们实验室的研究 证明H2S通过对线粒体能量学的双重影响（即 在低浓度下抑制电子通量，并抑制它。外源H2S的最高接触 发生在结肠（0.2-2.4 mm）中，并源自肠道居民菌群，该居民合成硫化物。初步的 我们实验室中的数据表明，结肠上皮细胞定量氧化物外源H2s对硫代硫酸盐，并且 鼠结肠细胞中硫化物氧化酶的定位和表达水平强烈 受硫酸盐的存在或不存在肠道中的细菌的影响。我假设动态 宿主和微生物硫代谢产物之间的相互作用会影响宿主代谢并影响寿命。使用 秀丽隐杆线虫是一种模型生物，它具有与涉及的直系同源基因的完整补体 H2S在人类中发现的生物发生和氧化，很容易被遗传和饮食操纵和 为了进行生命分析，我将通过解决以下两个目标来检验我的假设。 （i）我将表征如何 H2S水平的改变和反应性硫物种会影响有机氧化还原和Hisstone修饰 与秀丽隐杆线虫中的寿命和压力反应有关。 H2S水平及其氧化副产品将被调制 通过外源H2S，宿主硫化物氧化酶的RNA干扰敲低以及饮食，使用 大肠杆菌在参与硫代谢的特定基因中缺失。我将评估外源 与膳食H2S调制相对于遗传编码的还原氧化会影响有机氧化还原 敏感的GRX1-ROGFP2传感器和轨迹变化组蛋白甲基化和乙酰化状态，并报告 链接到氧化还原，H2S的可用性，饮食和衰老。 （ii）我将研究外源H2S暴露的差异 来自空气与肠道微生物影响宿主硫化物氧化酶的表达和定位，即 反应性硫的抽象和类型及其对寿命的影响。我将通过 代谢组和转录组分析，以识别受H2S暴露影响的途径。这 这些研究的成功完成将是我们理解硫的相互作用的基础 宿主微型界面的代谢。