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.

项目概要 宿主相关微生物组对皮质类固醇的代谢与宿主外周组织的代谢相当。边- 皮质类固醇的链代谢能够改变类固醇的功能类别。当前的重点 该项目正在阐明参与（目标 1）皮质醇侧链裂解和还原的微生物酶 17-酮基产生11-氧雄激素（目标2）21-脱羟基产生11-氧孕酮衍生物。 这些途径很重要，因为它们被假设会增强与人类使命相关的疾病。 NIH。推测肠道和泌尿道中 11-氧雄激素的形成会影响宿主免疫 功能，并且可能是前列腺癌的危险因素。 11-氧孕酮衍生物的形成 已被证明通过涉及“脱氢酶”的完善机制具有促高血压作用 假设'。我们的初步数据表明我们已经建立了侧链裂解酶， 类固醇-17,20-去氨酶，来自肠道和尿路细菌。最终产品 17-酮类固醇可以 还原为睾酮或表睾酮衍生物。我们培养了编码厌氧菌 用于（表）睾酮形成的酶，我们的初步数据证明了我们的专业知识和可行性 定位和表征这些酶。我们还展示了一种能够产生皮质类固醇 21- 的培养厌氧菌 脱羟基并提出了假设和初步数据，假定皮质类固醇脱羟基 (csd) 基因簇编码 21-脱羟基。一旦找到，我们将应用先前建立的生物信息学 揭示编码同源甾醇生物组酶的宿主相关细菌多样性的方法 （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甾醇生物组 具有上述相同功能的酶，我们将利用无偏见的功能宏基因组方法。 我们提供了额外可靠的初步数据，证明我们研究的可行性，包括（1） 收集 180 份粪便样本进行功能宏基因组筛选 (2) 利用 RNA-Seq 进行类固醇- 诱导基因的发现（3）众多微生物甾醇生物组的异源表达和表征 (3)酶测定法和类固醇代谢组学的建立。本文提出的研究 我们认为，该应用程序是创新的，因为它代表了与现状的新的实质性背离 四十年前通过结合转录组学、异源蛋白质表达、酶而建立的现状 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 筛选以发现编码皮质类固醇侧链代谢的精确核酸序列 酶。成功完成将加强对肠道之间因果关系的研究 固醇生物组和宿主疾病，并有望导致转化方法（益生菌、酶 抑制剂）来调节肠道微生物组以恢复和维持健康。