The Interactive Metabolome of Polymicrobial Biofilms

描述(申请人提供)：这是一份K01奖项的申请书，授予Vanessa Phelan博士，她是加州大学圣地亚哥分校斯卡格斯药学和药学学院药理学系的化学家。在多菌生物膜代谢组学的研究中，费兰博士确立了自己作为年轻研究员的地位。这一K01奖项将为Phelan博士提供必要的支持，以实现以下目标：(1)成为阐明复杂微生物群落中的化学串扰的专家；(2)对与囊性纤维化肺部感染相关的多菌生物膜内的复杂代谢交换进行临床相关研究；(3)采用最先进的质谱学技术来研究混合生物膜；(4)建立生物膜内微生物和代谢物的因果预测模型；以及(5)发展独立的研究事业。为了实现这些目标，Phelan博士组建了一个指导团队，成员包括主要导师、UCSD斯卡格斯药学和制药科学学院副教授Pieter Dorrestein博士，他负责微生物代谢交换的学术研究；以及一位共同导师：Suzanne Noble博士，她是加州大学旧金山分校医学系的助理教授，其在微生物遗传学方面的专业知识专注于白念珠菌、共分离株和哺乳动物宿主之间的相互作用。众所周知，微生物为种内和种间的相互作用产生小分子。然而，这些代谢物在更大的多微生物生物膜群落中的作用仍未得到研究。Phelan博士的研究将集中于确定负责建立和维持囊性纤维化肺部感染共分离物衍生的多微生物生物膜群落的代谢物(目标1)、通过基因敲除或引入亚抑制浓度的抗生素去除关键代谢物来扰乱这些群落的效果(目标2)以及开发生物体的预测分子网络(目标3)。费兰博士将使用最先进的质谱仪，包括微生物菌落的二维和三维成像质谱学，以及一种能够直接从培养皿中采集微生物菌落的新技术。这项研究将为微生物生物膜的多重研究奠定基础，包括在K奖结束前提出的R01拨款申请中提出的蛋白质组、转录组和代谢组相互作用。 公共卫生相关性：更好地理解人类致病有机体与其环境之间潜在的相互作用，有助于洞察疾病的基本原因。在这个项目中，我们将确定病原体用来与环境相互作用的分子。

DESCRIPTION (provided by applicant): This is an application for a K01 award for Dr. Vanessa Phelan, a chemist in the Department of Pharmacology at Skaggs School of Pharmacy and Pharmaceutical Sciences at the University of California, San Diego. Dr. Phelan is establishing herself as a young investigator in research of the metabolomics of polymicrobial biofilms. This K01 award will provide Dr. Phelan with the support necessary to accomplish the following goals: (1) to become expert at elucidating the chemical crosstalk in complex microbial communities; (2) to conduct clinically relevant investigations of the complex metabolic exchange within polymicrobial biofilms associated with cystic fibrosis lung infections; (3) to implement state-of-the-art mass spectrometry techniques to investigate mixed biofilms; (4) to develop a causal predictive model of the organisms and metabolites within biofilms and (5) to develop an independent research career. To achieve these goals, Dr. Phelan has assembled a mentoring team comprised of a primary mentor, Dr. Pieter Dorrestein, Associate Professor in Skaggs School of Pharmacy and Pharmaceutical Sciences, UCSD, who conducts academic research into metabolic exchange of microorganisms and one co-mentor: Dr. Suzanne Noble, an Assistant Professor in the Department of Medicine at University of California, San Francisco whose expertise in microbial genetics focuses her work on the interactions between C. albicans, co-isolates and the mammalian host. It is well established that microorganisms produce small molecules for both intra-species and inter- species interactions. However, the roles of these metabolites in the larger polymicrobial biofilm community remain uninvestigated. Dr. Phelan's research will focus on defining the metabolites responsible for establishing and maintaining a polymicrobial biofilm community derived of co-isolates from cystic fibrosis lung infections (Aim 1), the effect of perturbing these communities by removing key metabolites via gene knock-outs or introducing sub-inhibitory concentrations of antibiotics (Aim 2) and developing a predictive molecular network of the organisms (Aim 3). Dr. Phelan will use state-of-the-art mass spectrometry including two-dimensional and three-dimensional imaging mass spectrometry of microbial colonies and a new technique capable of sampling a microbial colony directly from a petri-dish. This research will form the basis for a multiplexed study of microbial biofilms includig proteomic, transcriptomic and metabolomic interactions to be proposed in an R01 grant application before the end of the K award. PUBLIC HEALTH RELEVANCE: Improved understanding of the underlying interactions between human disease causing organisms and their environment provides insight into the basic cause of disease. In this project we will identify the molecules that pathogens use to interact with their environment.