Characterization of bacterial reductases acting on the A-ring of 11-oxy-androgens

作用于 11-氧雄激素 A 环的细菌还原酶的表征

• 批准号: 10653436
• 负责人: Jason Michael Ridlon
• 金额: $ 3.59万
• 依托单位: UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-04-01 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10653436
• 关键词: Adrenal Cortex Hormones; African-American Graduate Student; Anaerobic Bacteria; Androgens; Animals; Bacteriology; Biochemistry; Bioinformatics; CRISPR/Cas technology; Clostridium; Development; Engineering; Enzymes; Escherichia coli; Feces; Female; Gene Expression; Genes; Glucocorticoids; Gnotobiotic; Grant; Human; Human Microbiome; Hydrocortisone; Immune; Immunology; Intestines; Laboratories; Link; Literature; Manuscripts; Mentors; Metabolism; Microbiology; National Institute of General Medical Sciences; Oxidoreductase; Parents; Research Personnel; Side; Steroids; Suspensions; Training; Writing; career development; gastrointestinal function; gene discovery; gut bacteria; gut microbiome; instructor; microbial; novel; nutrition; parent grant; steroid metabolism; training opportunity; transcriptome sequencing; transcriptomics

Project Summary/Abstract Literature from as early as 1957 from Nabarro et al demonstrated that glucocorticoids such as cortisol can be converted into potent androgens like hydroxyandrostenedione (11β-OHAD) (1,2). However, it wasn't until 1981 that side-chain cleavage of cortisol was first observed in human stool suspensions (3). In 1984, researchers isolated the human gut bacterium, Clostridium scindens, capable of steroid-17,20-desmolase (SD) (4,5). Ridlon et al (2013) utilized RNA-Seq to identify the cortisol-inducible genes (desAB), which were later confirmed by our laboratory to encode SD (6,7). The parent R01 seeks to study the contribution of both human gut bacteria expressing DesAB as well as engineered E. coli expressing DesAB to circulating 11β-OHAD, and the effect of this 11-oxy-androgen precursor on intestinal immune profile and gene expression. The aim of the proposed diversity supplement is to extend the studies in the parent grant by characterizing human microbiome genes encoding enzymes capable of further metabolism of 11β-OHAD into dihydro-stereoisomeric derivatives 11β- OH-5α-dihydroandrostane (androgen-precursor) and 11β-OH-5β-dihydroandrostane (non-androgen precursor) and integrating expression of these genes into gnotobiotic animal studies. These mechanistic studies represent the first attempt to causally link gut microbial corticosteroid metabolism to circulating 11-oxy-androgen levels and gastrointestinal function. This unique training opportunity integrates anaerobic bacteriology, microbial gene discovery and enzyme characterization, CRISPR-Cas9 recombineering, gnotobiology, immunology, host and microbial transcriptomics (RNA-Seq). Training will also consist of diverse career development and didactic coursework including NUTR 550 Grant Writing Course (Ridlon co-instructor) and one-on-one grant and manuscript writing development by the co-mentors, bioinformatics and statistical courses and HPCBio modules on transcriptomic analysis, nutrition, microbial and mammalian biochemistry courses, and microbiology.

项目总结/摘要 早在1957年，Nabarro等人的文献就证明皮质醇等糖皮质激素可以 转化为有效的雄激素，如羟基雄烯二酮（11β-OHAD）（1，2）。然而直到1981年 皮质醇的侧链裂解首先在人粪便悬浮液中观察到（3）。1984年，研究人员 分离出了能够产生类固醇-17，20-碳链酶（SD）的人类肠道细菌Clostridium plandens（4，5）。里德隆 等人（2013）利用RNA-Seq鉴定皮质醇诱导基因（desAB），后来我们的研究证实了这一点。 实验室编码SD（6，7）。亲本R 01试图研究人类肠道细菌和 表达DesAB以及工程化E.表达DesAB的大肠杆菌对循环11β-OHAD的影响，以及 这种11-氧雄激素前体对肠道免疫谱和基因表达的影响。建议的目的 多样性补充是通过表征人类微生物组基因来扩展父母资助的研究 编码能够将11β-OHAD进一步代谢为二氢立体异构衍生物11β- OH-5α-二氢雄烷（雄激素前体）和11β-OH-5β-二氢雄烷（非雄激素前体） 并将这些基因的表达整合到知菌动物研究中。这些机制研究代表了 首次尝试将肠道微生物皮质类固醇代谢与循环11-氧-雄激素水平因果联系起来， 胃肠功能这种独特的培训机会整合了厌氧细菌学，微生物基因 发现和酶表征，CRISPR-Cas9重组工程，基因生物学，免疫学，宿主和 微生物转录组学（RNA-Seq）。培训还将包括各种职业发展和教学 课程包括NUTR 550补助金写作课程（Ridlon联合讲师）和一对一补助金， 共同导师撰写的手稿，生物信息学和统计学课程以及HPCBio模块 转录组学分析、营养学、微生物和哺乳动物生物化学课程以及微生物学。