Collaborative Research: OSIB: Chemical signaling in a host-microbe symbiosis

合作研究：OSIB：宿主-微生物共生中的化学信号传导

• 批准号: 2220511
• 负责人: Mark Mandel
• 金额: $ 26.06万
• 依托单位: University of Wisconsin-Madison
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-15 至 2025-07-31
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2220511&HistoricalAwards=false
• 关键词: Collaborative; Research; OSIB; Chemical; signaling

Despite an appreciation of the importance of symbiotic microbes, the chemical language used between hosts and microorganisms is poorly understood. There is a current gap in knowledge when it comes to the expressed chemistry that bacteria utilize to adapt to host environments, and the identification of the chemical matter responsible would add significantly to our understanding of how bacteria are able to colonize and maintain themselves in niche environments or out-compete other microorganisms for the same niche. Our proposal is highly interdisciplinary, as is our team, incorporating forward genetics, cutting-edge analytical measurement science, and medicinal chemistry to identify how the glowing bacterium, Vibrio fischeri, uses a small chemical compound to better colonize its squid host while also discovering other molecules that have yet to be identified. Our system thus provides an entry point to identify chemical signals that may be relevant in other beneficial and pathogenic colonization systems and to elucidate the relevant targets in an accessible model. We will leverage our tractable, environmentally intriguing model system to create a hands-on outreach workshop with young girls in grades 5-10 in the greater Monterey Bay area, which has a high Latinx population, in collaboration with Expanding Your Horizons.There is significant interest in identifying and characterizing small molecules that underlie host-microbe communication in plant, animal, and human host-microbiome interactions. The naturally simple association between Euprymna scolopes bobtail squid and Vibrio fischeri luminescent bacteria provides a symbiotic relationship that can be interrogated in situ to discover functional compounds. Bacterial colonization of the host is accessible to sophisticated bacterial genetic and genomic approaches, and the site of infection can be imaged in live animals. Our collaborative efforts have developed novel methods to identify and study small molecules in this symbiosis. This work has resulted in the identification of a small organic molecule, termed diketopiperazine (DKP), as a potential contributor to symbiotic biofilm and luminescence phenotypes from V. fischeri. We have used imaging mass spectrometry to identify additional biofilm-associated molecules, and, using structure elucidation and synthetic chemistry, we have synthesized large amounts of the DKP. Here, we will leverage our interdisciplinary expertise in imaging mass spectrometry, structure isolation and elucidation, forward genetic screening, and synthetic bioorganic chemistry to identify the function of this DKP and identify the biosynthetic origin of this molecule to explore whether it is used in other microbial-host systems. This model system will allow us to address current knowledge gaps in our understanding of the small molecules that symbiotic bacteria produce during colonization of host specific tissues.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

尽管共生微生物的重要性得到了重视，但宿主和微生物之间使用的化学语言却知之甚少。当涉及到细菌用来适应宿主环境的表达化学时，目前的知识存在差距，并且负责的化学物质的鉴定将大大增加我们对细菌如何能够在生态位环境中定植和维持自己或竞争其他微生物的理解。我们的建议是高度跨学科的，我们的团队也是如此，结合了前沿遗传学，尖端的分析测量科学和药物化学，以确定发光细菌费氏弧菌如何使用一种小的化学化合物来更好地殖民其鱿鱼宿主，同时还发现了其他尚未确定的分子。因此，我们的系统提供了一个切入点，以确定化学信号，可能是相关的其他有益的和致病性的殖民系统，并阐明在一个可访问的模型中的相关目标。我们将利用我们的易处理的，环境有趣的模型系统，创建一个动手推广研讨会，与大蒙特雷湾地区的5-10年级的年轻女孩，其中有很高的拉丁裔人口，与扩展你的视野合作。有识别和表征小分子的兴趣，在植物，动物和人类宿主微生物组相互作用的宿主微生物通信的基础。Euprymna nopes bobtail squid和Vibrio fischeri发光细菌之间的自然简单关联提供了一种共生关系，可以原位询问以发现功能化合物。宿主的细菌定殖可通过复杂的细菌遗传学和基因组学方法获得，并且感染部位可在活体动物中成像。我们的合作努力已经开发出新的方法来识别和研究这种共生关系中的小分子。这项工作已经导致了一个小的有机分子，被称为二酮哌嗪（DKP），作为一个潜在的贡献共生生物膜和发光表型从费氏弧菌的鉴定。我们已经使用成像质谱法来识别额外的生物膜相关分子，并且使用结构解析和合成化学，我们已经合成了大量的DKP。在这里，我们将利用我们在成像质谱，结构分离和阐明，正向遗传筛选和合成生物有机化学方面的跨学科专业知识来确定这种DKP的功能，并确定这种分子的生物合成来源，以探索它是否用于其他微生物宿主系统。这个模型系统将使我们能够解决目前的知识差距，我们的理解的小分子，共生细菌在宿主特定tissues.This定殖过程中产生的使命反映了NSF的法定使命，并已被认为是值得的支持，通过评估使用基金会的智力价值和更广泛的影响审查标准。