Metabolomic Analysis as a Tool to Understanding the Use of Novel Therapeutics in

描述（由申请人提供）：我的长期职业目标是建立一个独立的研究生涯，研究细菌生物膜在伤口环境中介导对宿主先天免疫细胞的特异性病理作用，导致偏离正常伤口愈合过程并导致伤口慢性的假设。我在先天免疫学和医学生物膜方面的背景为我提供了独特的专业知识，使我能够提出有关免疫细胞-细菌生物膜相互作用的创新和基本问题。作为一名分子方法和细胞生物学的研究生，我的研究训练也给了我技术技能，使我能够将这些工具应用于慢性伤口模型的系统生物学分析；目前，我的目标是通过获得核磁共振和质谱代谢组学分析方面的专业知识来完成我的培训，这是解决宿主-病原体相互作用等系统生物学问题的重要方法。我的职业发展计划的目标是发展专业知识并掌握必要的分析工具，将综合代谢组学分析整合到免疫细胞对细菌生物膜暴露反应的全球系统生物学研究中。为了实现这个职业发展奖项的目标，我组建了一个导师团队，他们既有专业知识，可以在我感兴趣的技术技能方面对我进行培训，也有经验，可以成为有效的导师。德拉茨博士有近45个

DESCRIPTION (provided by applicant): My long-term career goal is to establish an independent research career addressing the hypothesis that bacterial biofilms mediate specific pathological effects against host innate immune cells within the wound environment which result in deviations from the normal wound healing process and lead to wound chronicity. My background in innate immunology and medical biofilms provides me with unique expertise enabling me to ask innovative and fundamental questions regarding immune cell-bacteria biofilm interactions. My research training as a graduate student in molecular methods and cell biology has also given me technical skills that will enable me to apply such tools for systems biology analysis of the chronic wound models; at present, I aim to complete my training by gaining expertise in NMR and MS metabolomic analysis, an essential approach to solving systems biology problems such as host- pathogen interactions. My goal with this career development plan is to develop the expertise and master the analytical tools necessary to integrate comprehensive metabolomics analyses into a global systems biology study of immune cell responses to bacterial biofilm exposure. To address my objectives for this career development award, I have assembled a mentorship team with both the expertise to train me in the technical skils of interest and the experience to be effective mentors. Dr. Dratz has nearly 45 years of experience as a NIH supported researcher making him an excellent choice as Senior Mentor of my team. In the course of this award, my objective is to acquire expertise in nuclear magnetic resonance (NMR), mass spectrometry (MS), in silico metabolic modeling, and chemometric analysis working closely with Drs. Copi¿, Bothner, and Carlson all experts in their respective scientific disciplines. In adition to acquiring technical skils necessary to undertake te proposed metabolomics studies, I will seek out professional development. To that end, my career development plan includes participation in educational opportunities such as guest lecturing and the mentorship of a graduate student, participation in training for the Responsible Conduct of Research, and engagement in the larger scientific community through participation in conferences, publication, and the peer-review process of grants and manuscripts. Montana State University (MSU) provides an excellent environment for this training with facilities and equipment that has been acquired within the last few years to develop a state-of-the-art Metabolomics/Systems Biology Research Center, including access to the expertise of core facility managers in NMR, MS, and bioinformatics. In addition, opportunities for intellectual stimulation abound including the Systems Biology Journal Club and cross-disciplinary research. The immediate objective of this career development plan is not only to acquire the technical expertise outlined above, but also to apply that training to the establishment of my own research program. My preliminary work led me to the hypothesis that the interface between innate immune cells and bacterial biofilms result in distinct metabolic profiles that can be manipulated for therapeutic treatment and perhaps can also be used for diagnostics. To assess the validity of this hypothesis, I propose to establish that the biofilm mode of growth of the opportunistic chronic wound pathogen P. aeruginosa results in distinct metabolic patterns and that the biofilms are especially sensitive to iron deprivation by the immune molecule lactoferrin, document that exposure to P. aeruginosa biofilms in vitro results in a metabolic deviation in innate immune cells as part of a phenotypic shift towards inflammation, and establish that introduction of lactoferrin to the in vitro host-pathogen chronic wound model results in metabolic starvation of the pathogen while shifting the innate immune cells toward a resident macrophage phenotype that more efectively resolves inflammation allowing the wound to progress to resolution. The studies proposed here have the potential to uncover mechanisms at the root of deviations from the normal healing process that result in the development of chronic wounds, and will provide molecular knowledge that may be used in the long term to develop novel therapeutic paths by the manipulation of metabolic pathways that control immune cell phenotype.