Discovery of GPCR-active natural products and their biosynthetic genes from the human associated bacteria

项目概述：以人类微生物群为灵感的疗法的开发至少在一定程度上是有限的 由于我们缺乏对人类相关(HA-)细菌如何与其人类宿主和 影响病原体。人类微生物组测序研究表明， 细菌种群与人类健康。尽管有这些相关性和将HA细菌与 小鼠的疾病，HA细菌如何具体影响哺乳动物生理的机制细节仍然存在 很大程度上是未知的。在其他环境中，已知细菌严重依赖低分子量。 化合物(小分子或天然产物)与其他生物体相互作用。同样，我们预计 HA细菌可能会使用小分子与其人类宿主和病原体相互作用。安装 有证据表明，尽管每个人类微生物群都由一组复杂的细菌组成，但一个 数量少得多的物种在大多数个体中非常普遍。虽然我们不知道 我们推测，究竟哪些HA细菌负责维持人类健康或导致疾病 这些常见的HA细菌产生的小分子可能会发挥重要作用 在这些过程中。这项建议的中心目标是筛选最常见的 高通量生物活性筛选中观察到的HA-细菌鉴定GPCR和SARS-CoV-2活性小分子 分子及其产生生物合成基因簇(BGC)。GPCR构成了最大的 真核细胞跨膜受体。众所周知，它们在人类生物学中扮演着不同而深远的角色 并且容易受到小分子的调节。基于GPCRs在以下方面发挥如此广泛的作用 在真核细胞中将环境中的化学信息转化为生物信号，我相信 HA-细菌可能通过产生与GPCRs相互作用的小分子来影响宿主生理。 SARS-CoV-2病毒的出现代表着一种没有治疗药物的世界性大流行 治疗。这项建议的两个目的将导致(1)鉴定、分离和结构阐明 HA-细菌编码的代谢物，可与不同的GPCR相互作用或抑制SARS-CoV-2，(2) 这些代谢物的基因簇的特征，以及(3)通过以下方式验证它们的产生 定植细菌。这些研究将有助于阐明HA-细菌如何形成的机制细节 人类健康。据报道，人类微生物群会影响复杂的病理生理过程 从免疫系统的调节到大脑和中枢神经系统的发育。 HA细菌种群的变化与影响超过2亿美国人的疾病有关 包括肥胖、糖尿病、炎症性肠病、自闭症、肠易激综合征和肝硬变等 还有很多其他人。从HA-细菌衍生的治疗方法在控制各种基础生物学方面具有潜在的实用价值 过程和人类疾病。

Project summary: The development of therapies inspired by the human microbiome is at least in part limited by our lack of understanding of how human associated (HA-) bacteria communicate with their human host and affect pathogens. Human microbiome sequencing studies show strong correlations between changes in bacterial populations and human health. Despite these correlations and the evidence linking HA-bacteria to disease in mice, the mechanistic details of how HA-bacteria specifically affect mammalian physiology remain largely unknown. In other environments, bacteria are known to rely heavily on low molecular weight compounds (small molecules or natural products) to interact with other organisms. Similarly, we expect that HA-bacteria are likely to use small molecules to interact with their human hosts and pathogens. Mounting evidence suggests that, although each human microbiome is composed of a complex collection of bacteria, a much smaller number of species is highly prevalent across the majority of individuals. While we don’t know exactly which HA-bacteria are responsible for maintaining human health or causing disease, we hypothesize that small molecules produced by these commonly encountered HA-bacteria are likely to play an important role in these processes. The central aim of this proposal is to screen metabolites produced by the most commonly observed HA-bacteria in high-throughput bioactivity screening to identify GPCR and SARS-CoV-2-active small molecules and their producing biosynthetic gene clusters (BGCs). GPCRs constitute the largest family of eukaryotic trans-membrane receptors. They are known to play diverse and profound roles in human biology and are prone to regulation by small molecules. Based on the fact that GPCRs play such an extensive role in transforming chemical information from the environment into biological signals in eukaryotic cells, I believe that HA-bacteria likely affect host physiology through the production of small molecules that interact with GPCRs. The emergence of the SARS-CoV-2 virus represents a worldwide pandemic with no therapeutic drug treatments. The two Aims of this proposal will result in (1) the identification, isolation, and structure elucidation of HA-bacteria-encoded metabolites that either interact with diverse GPCRs or inhibit SARS-CoV-2, (2) the characterization of the gene clusters for these metabolites, and (3) the validation of their production by colonizing bacteria. These studies will help to illuminate the mechanistic details of how HA-bacteria shape human health. The human microbiome is reported to influence complex pathophysiological processes ranging from the regulation of the immune system to the development of the brain and the central nervous system. Changes in HA-bacterial populations are associated with diseases that affect over 200 million Americans including obesity, diabetes, inflammatory bowel disease, autism, irritable bowel syndrome, and cirrhosis among many others. Therapies derived from HA-bacteria have potential utility in controlling diverse basic biological processes and human diseases.