Identifying the Human Targets of Secreted M. tuberculosis Effectors by Proteomics

描述（由申请人提供）：这是一份申请指导临床科学家研究职业发展奖（K08）的申请，申请对象是Bennett Penn博士，他是加州大学旧金山分校传染病部的三年级研究员，即将被任命为医学教授（兼职系列）。他的长期目标是确定结核分枝杆菌引起人类疾病的分子机制，并根据这些信息开发新的治疗方法。该应用程序的具体目标是使用质谱（MS）和遗传方法的组合来识别结核分枝杆菌毒力因子靶向的功能重要的人类蛋白质，并为Penn博士获得必要的额外培训，以领导他自己的研究小组。在他的初步研究中，他使用亲和纯化和质谱（MS）鉴定了结核分枝杆菌与人类细胞之间数百种新的蛋白质-蛋白质相互作用。在目标1中，他将使用一套生物信息学和生化方法来完善和验证这些MS发现。在目标2中，他将使用遗传方法来确定哪些相互作用在结核分枝杆菌感染期间发挥重要的功能作用。在Aim 3中，将对这些相互作用中的一部分进行详细的生化分析，以确定它们破坏宿主免疫功能的分子机制。佩恩博士组建了一个由三位共同导师组成的团队，他们在多个学科之间架起桥梁，指导他的持续进步。重要的是，由于Penn博士的博士工作是在哺乳动物发育生物学领域，K08奖将提供一段额外的研究培训，以获得操纵毒性结核分枝杆菌和进行MS实验的专业技能。K08还将通过UCSF提供的重点课程和研讨会，为获得管理独立研究小组的技能提供额外的支持，并将允许Penn博士在SFGH结核病诊所工作，以保持他的临床专业。获得该奖项后，佩恩博士将领导自己的研究小组，并提交一份R01，进一步扩展他对结核病中宿主-病原体相互作用的研究。

DESCRIPTION (provided by applicant): This is an application for a Mentored Clinical Scientist Research Career Development Award (K08) for Dr. Bennett Penn, a third-year fellow Division of Infectious Diseases with pending appointment as Professor of Medicine (adjunct series) at UCSF. His long-term goal is to determine the molecular mechanisms by which M. tuberculosis causes human disease and to develop new therapeutics based on this information. The specific goal of this application is to use a combination of mass spectrometry (MS) and genetic approaches to identify the functionally important human proteins targeted by M. tuberculosis virulence factors, and to obtain the necessary additional training for Dr. Penn to lead his own research group. In his preliminary studies, he has used affinity purification and mass spectrometry (MS) to identify several hundred novel protein-protein interactions between M. tuberculosis and human cells. In Aim 1, he will use a set of bioinformatics and biochemical approaches to refine and validate these MS findings. In Aim 2, he will use genetic approaches to determine which of these interactions play functionally important roles during M. tuberculosis infection. In Aim 3, a select number of these interactions will be subjected to detailed biochemical analysis to establish the molecular mechanisms by which they factors disrupt host immune function. Dr. Penn has assembled a team of three co-mentors that bridge several disciplines to guide his continued advancement. Importantly, since Dr. Penn's doctoral work was in the field of mammalian developmental biology, the K08 award will provide a period of additional research training to acquire the specialized skills for manipulating virulent M. tuberculosis, and carrying out MS experiments. The K08 will also provide added support for acquiring the skills to manage an independent research group through focused courses and seminars offered by UCSF, and will permit Dr. Penn to maintain his clinical specialization by working at the SFGH TB Clinic. By the completion of this award Dr. Penn will be in an excellent position lead his own research group and to submit an R01 that further extends his studies on host-pathogen interactions in TB. RELEVANCE: This project is relevant to human health because understanding how Mycobacterium tuberculosis disrupts the immune system will lay the foundation for developing therapies aimed at restoring these functions and thereby improving therapy for tuberculosis.