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周围组织的代谢相媲美。侧面- 皮质类固醇的链式代谢能够改变类固醇的功能类别。当前的焦点 该项目正在阐明(目标1)皮质醇侧链断裂和还原所涉及的微生物酶 17-酮产生11-氧基-雄激素(目标2)21-脱羟基得到11-氧基孕酮衍生物。 这些途径具有重要意义，因为它们被假设为增强了与 国家卫生研究院。肠道和尿路中11-氧-雄激素的形成被认为影响宿主免疫 功能，并可能是前列腺癌的危险因素。11-氧基孕酮衍生物的形成 已经被证明是通过与脱氢酶有关的一种公认的机制来促进高血压的 假设‘。我们的初步数据表明，我们已经建立了侧链裂解酶， 肠道和尿路细菌中的类固醇-17，20-脱解酶。最终产物17-酮类固醇可以是 还原为睾丸素或表雄酮衍生物。我们培养的厌氧菌编码了 睾丸激素形成的酶，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，并提出了一个假说和初步数据，推测皮质类固醇脱羟基 (CSD)基因簇编码21-脱羟基。一旦定位，我们将应用先前建立的生物信息学 揭示编码同源类固醇生物体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，为了解决进化上相似的类固醇生物体 具有上述相同功能的酶，我们将利用无偏见的功能元基因组学方法。 我们提供了更多稳健的初步数据，以证明我们研究的可行性，这些研究包括(1) 收集180份粪便样本用于功能元基因组筛选(2)利用RNA-SEQ检测类固醇 可诱导基因发现(3)多种微生物甾体生物体的异源表达与鉴定 酶类(3)酶测定和类固醇代谢组学的建立。这项研究中提出的 我们认为，申请是创新的，因为它代表了对现状的新的实质性偏离 四十年前通过结合转录学、异源蛋白表达、酶 分析、系统发育和序列相似性网络、合成生物学和功能元基因组学 筛选发现编码糖皮质激素侧链代谢的精确核酸序列 酵素。成功的完成将加强对肠道之间因果关系的研究 类固醇生物体和宿主疾病，并有望导致翻译方法(益生菌、酶 抑制物)调节肠道微生物群，以恢复和维持健康。