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.

在内分泌学文献中存在着一个主要的争议。羟基染色器（11β-OHAD）。认为11β-OHAD的合成主要是在通过雄酮的11β-羟基化肾上腺（A4）。另一个路线是穿过侧链裂解皮质醇的含量，尽管被认为很小。患有17-羟化酶/C17,20-裂解酶（CYP17A1）的患者缺乏症患者无法合成皮质醇或A4。当外源给予皮质醇时，尿道表现出衍生物 11β-OHAD。这暗示了一种新型的宿主酶，或者是负责侧链的肠道微生物酶皮质醇的切割。我们已经在高度活跃的肠道微生物途径中鉴定了可能是负责的基因用于皮质醇的侧链切割。我们假设肠道菌群很重要，并且被理解宿主内分泌系统的成分，该系统产生大量的11β-OHAD。这个项目将通过比较无菌猪和猪的稳定同位素标记的皮质醇代谢来检验该假设具有不同复杂性的肠道微生物组。我们将通过单一来确定对宿主结肠生理的影响免疫细胞的细胞RNA-seq，免疫组织化学和流式细胞仪。了解身体角色肠道细菌的侧链裂解对于确定未来的策略也很重要雄激素形成。我们将在体外确定对皮质醇和11β-OHAD的细菌转录组反应和体内。此外，要证明负责皮质醇侧链的基因之间的原因裂解（DESAB）和主机类固醇代谢组概况，我们将使用合成生物学方法来工程 Desab途径（野生型和非活性突变体）进入大肠杆菌，并定居Gnotobiotic Pigs。这些预计研究将解决已经存在数十年的谜团，如果它可能导致范式偏移可以证明，肠道微生物群对数量上主要的宿主类固醇和前雄激素。