Gut bacterial metabolism of the side-chain of corticosteroids

皮质类固醇侧链的肠道细菌代谢

• 批准号: 10703384
• 负责人: Jason Michael Ridlon
• 金额: $ 36.08万
• 依托单位: UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-15 至 2026-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10703384
• 关键词: Address; Adrenal Cortex Hormones; Affect; Amino Acid Sequence; Anaerobic Bacteria; Androgens; Androstenedione; Animal Model; Bacteria; Base Sequence; Biochemical; Bioinformatics; Biological Assay; Collection; Colon Carcinoma; Communities; Cytochrome P450; Data; Disease; Enzyme Inhibitor Drugs; Enzymes; Escherichia coli; Feces; Funding; Future; Gene Cluster; Genes; Glucocorticoids; Goals; Health; Health Promotion; Homologous Gene; Human; Human Microbiome; Hydrocortisone; Hypertension; Intestines; Ketones; Ketosteroids; Knowledge; Libraries; Malignant neoplasm of prostate; Maps; Metabolic Biotransformation; Metabolism; Metagenomics; Methodology; Mission; Mixed Function Oxygenases; Organism; Oxidoreductase; Oxygen; Pathway interactions; Peripheral; Phylogenetic Analysis; Physiology; Probiotics; Process; Progesterone; Reaction; Research; Risk Factors; Side; Steroids; Testosterone; Therapeutic; Tissues; Transketolase; United States National Institutes of Health; Urinary tract; bacterial metabolism; bile acid metabolism; biobank; dehydroxylation; gene discovery; gut bacteria; gut microbiome; host-associated microbial communities; host-microbe interactions; hydroxyl group; hypertensive; immune function; innovation; metabolome; metabolomics; metagenome; microbial; microbiome; novel; prevent; protein expression; protein folding; screening; steroid metabolism; stool sample; synthetic biology; therapeutic target; transcriptome sequencing; transcriptomics; translational approach

Project Summary The metabolism of corticosteroids by host-associated microbiomes rivals that of host peripheral tissues. Side- chain metabolism of corticosteroids is capable of changing the functional class of steroid. The focus of the current project is elucidating the microbial enzymes involved in (Aim 1) the side-chain cleavage of cortisol and reduction of 17-keto resulting in 11-oxy-androgens (Aim 2) 21-dehydroxylation yielding 11-oxy-progesterone derivatives. These pathways are significant because they are hypothesized to potentiate diseases relevant to the mission of NIH. The formation of 11-oxy-androgens in the intestine and urinary tract is hypothesized to affect host immune function, and may be a risk factor for prostate cancer. The formation of 11-oxy-progesterone derivatives has been shown to be pro-hypertensive through a well-established mechanism involving the ‘dehydrogenase hypothesis’. Our preliminary data demonstrates that we have established the side-chain cleavage enzyme, steroid-17,20-desmolase, from both gut and urinary tract bacteria. The end-product 17-keto steroid can then be reduced to either testosterone or epitestosterone derivatives. We have cultured anaerobes that encode the enzymes for (epi)testosterone formation, and our preliminary data demonstrates our expertise and the feasibility of locating and characterizing these enzymes. We also show a cultured anaerobe capable of corticosteroid 21- dehydroxylation and formulated a hypothesis and preliminary data that a putative corticosteroid dehydroxylation (csd) gene cluster encodes 21-dehydroxylation. Once located, we will apply previously established bioinformatics approaches to uncover the diversity of host-associated bacteria encoding homologous sterolbiome enzymes (phylogenetic analysis, sequence similarity networks). However, to address evolutionarily analogous sterolbiome enzymes with the same functions described above, we will utilize an unbiased functional metagenomic approach. We present additional robust preliminary data demonstrating the feasibility of our studies that include (1) collection of 180 stool samples for functional metagenomic screening (2) utilization of RNA-Seq for steroid- inducible gene discovery (3) heterologous expression and characterization of numerous microbial sterolbiome enzymes (3) establishment of enzyme assays and steroid metabolomics. The research proposed in this application is innovative, in our opinion, because it represents a new and substantive departure from the status quo established four decades ago by combining transcriptomics, heterologous protein expression, enzyme assays, phylogenetic and sequence similarity networks, synthetic biology, and functional metagenomic screening to discover the precise nucleic acid sequences encoding corticosteroid side-chain metabolizing enzymes. Successful completion will potentiate research into the cause and effect relationships between the gut sterolbiome and host diseases, and is expected to lead to translational approaches (probiotics, enzyme inhibitors) to modulate the gut microbiome to restore and maintain health.

项目摘要 宿主相关微生物群对皮质类固醇的代谢与宿主外周组织的代谢相当。侧- 皮质类固醇的链代谢能够改变类固醇的功能类别。当前的焦点 一个项目是阐明微生物酶参与（目标1）侧链裂解皮质醇和还原 17-酮的11-氧-雄激素（Aim 2）21-脱羟基化产生11-氧-孕酮衍生物。 这些途径是重要的，因为它们被假设为增强疾病相关的使命， 国家卫生研究院11-氧雄激素在肠道和泌尿道的形成被假设影响宿主免疫 功能，并可能是前列腺癌的危险因素。11-羟-孕酮衍生物的形成具有 已被证明是促高血压通过一个完善的机制，涉及'脱氢酶 假设”。我们的初步数据表明，我们已经建立了侧链切割酶， 类固醇-17，20-碳链酶，来自肠道和泌尿道细菌。然后可以将终产物17-酮类固醇 还原为睾酮或表睾酮衍生物。我们培养的厌氧微生物 酶（表）睾酮形成，我们的初步数据表明我们的专业知识和可行性 来定位和鉴定这些酶。我们还展示了一种培养的厌氧菌，能够分泌皮质类固醇21- 并制定了一个假设和初步数据，即一个假定的皮质类固醇脱羟基 (csd)基因簇编码21-去羟基化。一旦定位，我们将应用先前建立的生物信息学 揭示宿主相关细菌编码同源甾醇生物组酶多样性的方法 （系统发育分析，序列相似性网络）。然而，为了解决进化上类似的固醇生物群系， 对于具有上述相同功能的酶，我们将利用无偏倚的功能宏基因组方法。 我们提出了额外的强有力的初步数据，证明我们的研究的可行性，包括（1） 收集180个粪便样本用于功能宏基因组筛选（2）利用RNA-Seq进行类固醇- 诱导基因发现（3）多种微生物甾醇生物组异源表达和表征 （3）酶测定法和甾体代谢组学的建立。这项研究提出， 我们认为，申请具有创新性，因为它代表着对地位的新的实质性偏离， 四十年前，通过将转录组学、异源蛋白表达、酶 分析、系统发育和序列相似性网络、合成生物学和功能宏基因组学 筛选以发现编码皮质类固醇侧链代谢的精确核酸序列 内切酶成功完成将加强对肠道之间因果关系的研究， 甾醇生物组和宿主疾病，并预计导致翻译方法（益生菌，酶 抑制剂）来调节肠道微生物组以恢复和维持健康。