Role of Gut Bacterial Side-Chain Cleavage of Cortisol in Host 11Beta-Hydroxyandrostenedione Formation

肠道细菌皮质醇侧链裂解在宿主 11β-羟基雄烯二酮形成中的作用

基本信息

批准号：
10726864
负责人：
Jason Michael Ridlon
金额：
$ 6.15万
依托单位：
UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-04-01 至 2025-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10726864
关键词：
Ablation Adaptive Immune System Adrenal Glands Affect Anabolism Androgen Receptor Androgens Androstenedione Androsterone Animals Antibiotics Bacteria Blood Circulation Bone Marrow CYP17A1 gene Carbohydrates Cells Chromosomes, Human, 16-18 Clinical Research Clostridium Colon Complex Corticosterone Dendritic Cells Disease Dose Endocrine system Endocrinology Engineering Enzymes Escherichia coli Etiology Excretory function Exhibits Family suidae Feces Flow Cytometry Future Gene Expression Gene Expression Profile Genes Germ-Free Glucocorticoids Gnotobiotic Goals Health Human Human Activities Hydrocortisone Hydroxylation Immune Immunohistochemistry In Vitro Infusion procedures Innate Immune System Literature Lyase Lymphoid Tissue Metabolism Microsomes Minor Mixed Function Oxygenases Modeling Mucous Membrane Mus Nuclear Oral Pathway interactions Patients Physiological Physiology Production Rattus Rectum Rodent Role Route Serum Site-Directed Mutagenesis Stable Isotope Labeling Steroids Surface Testing Tissues Urine Vitamins absorption cytokine dehydroepiandrosterone experimental study gastrointestinal gene discovery gut bacteria gut colonization gut microbiome gut microbiota immune function immune system function improved in vivo in vivo Model innovation intravenous injection mesenteric lymph node metabolome metabolomics microbial microbiome mutant novel porcine model rectal response single-cell RNA sequencing stable isotope synthetic biology transcriptome transcriptome sequencing transcriptomics urinary

项目摘要

A major controversy exists in the endocrinology literature regarding the biosynthesis of the pro-androgen 11β- hydroxyandrostenedione (11β-OHAD). The synthesis of 11β-OHAD is thought mainly to be synthesized in the adrenal gland by 11β-hydroxylation of androstenedione (A4). Another route is through the side-chain cleavage of cortisol, although it is thought to be minor. Patients with 17-hydroxylase/C17,20-lyase (CYP17A1) deficiency fail to synthesize either cortisol or A4. When given cortisol exogenously, the urinary profile exhibits derivatives of 11β-OHAD. This suggests either a novel host enzyme, or a gut microbial enzyme responsible for side-chain cleavage of cortisol. We have identified genes in a highly active gut microbial pathway that may be responsible for the side-chain cleavage of cortisol. We hypothesize that gut microbiota are an important, and understudied component of the host endocrine system, which generate significant quantities of 11β-OHAD. This project will test this hypothesis by comparing stable isotope-labeled cortisol metabolism in germ-free pigs and pigs colonized with gut microbiomes of different complexity. We will determine effects on host colonic physiology through single- cell RNA-Seq, immunohistochemistry, and flow cytometry of immune cells. Understanding the physiological role of side-chain cleavage by gut bacteria is also important in determining future strategies to modulate pro- androgen formation. We will determine bacterial transcriptomic responses to cortisol and 11β-OHAD both in vitro and in vivo. Furthermore, to demonstrate causation between the genes responsible for cortisol side-chain cleavage (desAB) and host steroid metabolome profile, we will utilize a synthetic biology approach to engineer the desAB pathway (both wild type and inactive mutant) into E. coli and colonize the gnotobiotic pigs. These studies are expected to resolve an enigma that has existed for decades, and may result in a paradigm-shift if it can be shown that the gut microbiota contributes significantly to a quantitatively major host steroid and pro- androgen.

内分泌学文献中关于前雄激素 11β- 的生物合成存在重大争议。羟基雄烯二酮 (11β-OHAD) 11β-OHAD 的合成被认为主要是在肾上腺通过雄烯二酮 (A4) 的 11β-羟基化作用的另一种途径是通过侧链裂解。皮质醇，尽管被认为是轻微的 17-羟化酶/C17,20-裂解酶 (CYP17A1) 缺乏症患者。无法合成皮质醇或 A4。当外源给予皮质醇时，尿液中会出现衍生物。 11β-OHAD 的存在，这表明要么是一种新的宿主酶，要么是一种负责侧链的肠道微生物酶。我们已经确定了高度活跃的肠道微生物途径中可能负责皮质醇裂解的基因。我们发现肠道微生物群是一个重要且尚未得到充分研究的因素。宿主内分泌系统的组成部分，产生大量的 11β-OHAD。通过比较无菌猪和定植猪的稳定同位素标记的皮质醇代谢来检验这一假设我们将通过单一的方法确定对宿主结肠生理学的影响。细胞 RNA 测序、免疫组织化学和流式细胞术了解免疫细胞的生理作用。肠道细菌对侧链的裂解对于确定未来调节亲的策略也很重要我们将在体外测定细菌对皮质醇和 11β-OHAD 的转录组反应。此外，还证明了负责皮质醇侧链的基因之间的因果关系。裂解（desAB）和宿主类固醇代谢组图谱，我们将利用合成生物学方法来设计 desAB 途径（野生型和失活突变体）进入大肠杆菌并定植于无菌猪中。研究预计将解决一个存在了数十年的谜题，如果它能够实现，可能会导致范式转变。可以证明，肠道微生物群对主要宿主类固醇和亲的数量有显着贡献。雄激素。