Role of the human gut microbiome and related metabolites for causally mediating vascular endothelial function

项目摘要/摘要 此F31申请的目的是为博士二年级博士生阿比盖尔·卡索女士提供支持。 道格拉斯·海尔斯(赞助商)在科罗拉多大学博尔德大学的实验室，进行研究和培训 这将使她做好准备，成为转化性心血管领域的独立研究员。 老龄化研究旨在预防和治疗年龄相关性心血管疾病(CVD)。作为她的一部分 拟议的培训计划，她的目标是完善目前正在开发的研究技能，并学习各种 新的技术、概念和专业技能，包括临床前小鼠模型的使用和获得 使用人类生物标本、细胞培养生物分析和代谢组学方法的新体验。她 拟议的研究项目旨在调查与年龄相关的变化在人类肠道微生物群中的作用。 以及循环肠道微生物衍生的代谢物损害血管内皮功能。肠道微生物群 成分随着心血管疾病和衰老而独特地改变，以及循环中与年龄相关的浓度的变化 某些肠源性代谢产物与心血管疾病有关。然而，这些与年龄相关的变化是否 原因是内皮功能受损尚不清楚。在强劲的初步数据指引下，卡索将确定： (目标1)人类肠道微生物群中与年龄相关的变化是否由于增加而损害内皮功能 使用创新的人源化小鼠模型和EX的氧化应激和一氧化氮生物利用度降低 活体“药物解剖”技术；(目标2A)与年龄相关的肠道微生物群诱导的 应用细胞培养生物测定法研究血浆循环因子对血管内皮细胞功能的影响 代谢物通过靶向代谢组学被老化的人体肠道微生物组改变；以及(目标3)如果特定 随着年龄增长，循环中的肠源性代谢产物直接导致内皮功能障碍。 细胞培养和分离血管的方法。预期的结果将确定与肠道微生物组相关的新的 和循环中的肠源性代谢物调节内皮功能，并可能促进这些 代谢产物作为防治年龄相关性内皮功能障碍的新治疗靶点。 总体而言，拟议的研究有可能解决重要的NHLBI战略愿景研究 优先事项，包括：1)调查对心血管疾病发病具有重要意义的新的病理生物学机制；2) 确定预防和治疗CVD113的新治疗靶点。海尔斯博士是一位国际公认的 美国国立卫生研究院资助的科学家，在翻译简历研究方面有成功指导的丰富历史，特别是在 “血管老化”这一新兴领域。在他的监督下，在内容专家的指导下 在维也纳·布伦特博士、安吉洛·达莱桑德罗博士和蒂凡尼·威尔的指导下，卡索女士将能够成功地 完成建议的研究和培训计划，促进她不断发展成为一名 独立从事机械式和平移式简历老化研究。

PROJECT SUMMARY/ABSTRACT The purpose of this F31 application is to provide support for Ms. Abigail Casso, a 2nd year PhD student in Dr. Douglas Seals’ (sponsor) laboratory at the University of Colorado Boulder, to conduct research and training that will prepare her to become an independent investigator in the field of translational cardiovascular (CV) aging research aimed at the prevention and treatment of age-related CV diseases (CVD). As part of her proposed training plan, she aims to both refine research skills presently under development and learn a variety of new technical, conceptual, and professional skills, including the use of preclinical mouse models and gaining new experiences using human biospecimens, cell culture bioassays, and metabolomics approaches. Her proposed research project seeks to investigate the role of age-related changes in the human gut microbiome and circulating gut microbiome-derived metabolites in impairing vascular endothelial function. Gut microbiome composition is uniquely altered with CVD and aging, and age-related changes in circulating concentrations of certain gut-derived metabolites have been related to CVD. However, whether these age-related changes causally impair endothelial function is unknown. Guided by strong preliminary data, Ms. Casso will determine: (Aim 1) If age-related changes in the human gut microbiome impair endothelial function due to increased oxidative stress and reduced nitric oxide bioavailability using innovative “humanized” mouse models and ex vivo “pharmaco-dissection” techniques; (Aim 2A) The effect of age-related, gut microbiome-induced changes in plasma circulating factors on endothelial function using cell culture bioassays; (Aim 2B) Which circulating metabolites are altered by the aging human gut microbiome via targeted metabolomics; and (Aim 3) If specific gut-derived metabolites that are increased in circulation with age directly induce endothelial dysfunction using cell culture and isolated vessel approaches. The expected results will identify novel gut microbiome-related and circulating gut-derived metabolite modulators of endothelial function and may facilitate translation of these metabolites as new therapeutic targets for prevention and treatment of age-related endothelial dysfunction. Overall, the proposed research has the potential to address important NHLBI Strategic Vision research priorities, including: 1) investigate new pathobiological mechanisms important to the onset of CVD; and 2) identify novel therapeutic targets to prevent and treat CVD113. Dr. Seals is an internationally recognized and NIH-funded scientist with a strong history of successful mentoring in translational CV research, particularly in the emerging field of “vascular aging”. Under his supervision and with the guidance of content expert co- mentors Drs. Vienna Brunt, Angelo D’Alessandro, and Tiffany Weir, Ms. Casso will be able to successfully complete the proposed research and training plan, facilitating her ongoing development towards becoming an independent investigator in mechanistic and translational CV aging research.