Exploring the endophytic parvome for the discovery of novel biologically active molecular scaffolds

探索内生细小组以发现新型生物活性分子支架

• 批准号: RGPIN-2019-04114
• 负责人: Gray, Christopher
• 金额: $ 2.11万
• 依托单位: University of New Brunswick
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2020
• 资助国家: 加拿大
• 起止时间: 2020-01-01 至 2021-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=714335
• 关键词: Exploring; endophytic; parvome; discovery; novel

The growing emergence of microbial strains that are resistant to currently used antibiotic agents is a major threat to human health as resistance to all of the major classes of antibiotics currently in clinical use has already been observed. Pan-resistant pathogens are increasingly being discovered and current forecasts predict that broad-scale ineffectiveness of antimicrobial agents is imminent. The discovery of novel natural products provides unparalleled access to unprecedented molecular scaffolds and unique molecular structures with desirable bioactivities and potencies that are unavailable through rational and combinatorial synthetic strategies. My research program focuses on the isolation of biologically active natural products that exhibit activity against pathogenic microbes, and consequently have significant potential as lead compounds for the development of new therapeutics for infectious disease. We focus on natural products produced by endophytic fungi of terrestrial, marine and freshwater plants, and a library of about 500 fungal endophyte cultures, many of which could represent new fungal species, is currently under investigation. Additional endophytic strains are continuously being isolated to increase our collection and we will also ferment fungi in co-culture with other fungi and bacteria and under various biological and chemical conditions to trigger cryptic biosynthetic genes and maximise the chemical diversity of our collection. My group has developed simple, robust and effective bioactivity and metabolomic profiling procedures that we will be applying in our search for novel molecular scaffolds. Fungal extracts will be evaluated in a suite of bioassays to assess their inhibitory effects against pathogenic microorganisms and their ability to modulate efflux pumps within these pathogens. This will allow us to compile bioactivity profiles for the extracts that can be related to the modes-of-action of their antibiotic constituents in an effort to discover new antibiotic classes and also identify natural products that can be used to increase the effectiveness of our current antibiotics by remediating for the effects of drug resistance. In conjunction with the compilation of the bioactivity profiles, NMR and LC-MS based metabolomics approaches will be used to dereplicate extracts that contain known antibiotics and prioritise projects for further investigation. Extracts that exhibit promising bioactivity profiles and unique metabolomic signatures will be subjected to bioassay-guided fractionation to isolate and identify the active components. The modes-of-action of the novel natural products that are isolated will then be probed by genomics, proteomics and chemical genetics. My research will ultimately lead to the discovery of lead compounds from the extensive biodiversity provided by fungal endophytes for the development of new therapeutics to fight infectious disease.

对目前使用的抗生素产生耐药性的微生物菌株的不断出现，对人类健康构成了重大威胁，因为已经观察到对目前临床使用的所有主要类别的抗生素都产生了耐药性。越来越多的泛耐药病原体被发现，目前的预测表明抗菌药物即将大规模失效。 新型天然产物的发现为获得前所未有的分子支架和独特的分子结构提供了无与伦比的途径，这些结构具有理想的生物活性和效力，而这是通过合理和组合合成策略无法获得的。我的研究项目重点是分离具有抗病原微生物活性的生物活性天然产物，因此具有作为开发传染病新疗法的先导化合物的巨大潜力。我们专注于陆地、海洋和淡水植物内生真菌产生的天然产物，目前正在研究约 500 种内生真菌培养物的库，其中许多可以代表新的真菌物种。我们不断分离出更多的内生菌株，以增加我们的收藏，我们还将在与其他真菌和细菌共培养的情况下，在各种生物和化学条件下发酵真菌，以触发神秘的生物合成基因，并最大限度地提高我们收藏的化学多样性。 我的团队开发了简单、强大且有效的生物活性和代谢组学分析程序，我们将应用这些程序来寻找新型分子支架。真菌提取物将通过一系列生物测定进行评估，以评估它们对病原微生物的抑制作用以及它们调节这些病原体内的外排泵的能力。这将使我们能够编制与其抗生素成分的作用模式相关的提取物的生物活性概况，以努力发现新的抗生素类别，并确定可用于提高我们当前抗生素有效性的天然产品。通过修复耐药性的影响来使用抗生素。结合生物活性概况的汇编，基于 NMR 和 LC-MS 的代谢组学方法将用于消除含有已知抗生素的提取物，并确定项目的优先顺序以供进一步研究。表现出有前景的生物活性特征和独特代谢组学特征的提取物将接受生物测定引导的分级分离，以分离和鉴定活性成分。然后，将通过基因组学、蛋白质组学和化学遗传学来探讨分离的新型天然产物的作用模式。我的研究最终将导致从真菌内生菌提供的广泛生物多样性中发现先导化合物，用于开发对抗传染病的新疗法。