Human genetic determinants of mycobacterial immunity

项目摘要/摘要 这个独立之路提案的主要目标是充分了解IL-12的作用 IL-23信号在人类分枝杆菌免疫中的作用。孟德尔人对分枝杆菌病的易感性 (MSMD)是一种遗传性疾病，儿童在受到弱毒力感染时容易患上严重疾病 分枝杆菌，包括卡介苗。尽管MSMD很少见，但对这种疾病的研究有能力 阐明对毒力更强的结核分枝杆菌免疫的基本免疫学机制。在……里面 在建议的研究方案中，我将使用从‘自然实验’中收集的知识，即先天错误 对分枝杆菌免疫功能的研究，加深我们对分枝杆菌发病机制的总体认识。尤其是，我 将从功能上表征我最近在IL12RB2和IL23R基因中发现的新的双等位基因突变 编码可能与IL-12Rβ1配对的受体链，这是细胞对IL-12和 IL-23(目标1)。这些突变存在于两个有多个MSMD感染儿童的家族中。 IL-12和IL-23的受体被认为是典型的Th1或Th17 T细胞的组成部分 并且已知在干扰素-γ的产生和/或IL-17的产生中发挥关键作用。因此，我会 进行详细的机制研究，以了解这些新的单基因损伤的不同影响 关于Th1和Th17细胞的发育和功能(目标2)。最后，我将测试假设中的缺陷 IL-12或IL-23信号导致这些患者的MSMD，并试图确定其他单基因原因 在500名其他MSMD患者中免疫相关基因中的MSMD(目标3)。在被指导的K99 在颁奖阶段，我将受益于Jean-Laurent博士独特的学习环境和临床资源 卡萨诺瓦在洛克菲勒大学(RU)的实验室。卡萨诺瓦博士的实验室是单基因研究的世界领先者 对传染病的解剖和对分枝杆菌的易感性的询问是该实验室时间最长的 项目。在卡萨诺瓦博士的实验室工作期间，我将享受与周围医生和患者无与伦比的接触 世界，以及许多经验丰富的同事的密切建议。我还将获得与尖端技术相关的经验 技术包括全外显子组和全基因组测序，RNAseq，成像流式细胞术，以及 多路细胞因子阵列。我职业发展计划中的一个关键部分是科学合作和职业生涯 我的咨询委员会提供的发展咨询意见，由卡萨诺瓦博士、路易吉·诺塔兰杰洛博士组成 (NIAID/NIH)和斯图尔特·唐耶博士(澳大利亚加文研究所)，他们都承诺在我的 在我独立的事业发展中提出了研究和建议。总之，拟议的计划将实现我的 阐述导致分枝杆菌易感性的单基因损伤的短期科学目标 我的长期目标是获得一个独立的角色，研究儿童的宿主基因决定因素 对病原体免疫。

Project Summary / Abstract The primary goal of this Pathway to Independence proposal is to gain a full understanding of the roles of IL-12 and IL-23 signaling in human mycobacterial immunity. Mendelian susceptibility to mycobacterial disease (MSMD) is a genetic condition predisposing children to severe disease upon infection by weakly virulent mycobacteria, including the BCG vaccine. Although MSMD is rare, the study of this disease has the power to elucidate the basic immunological mechanisms of immunity to the more virulent Mycobacterium tuberculosis. In the proposed research program, I will use knowledge gleaned from ‘natural experiments’, that is, inborn errors of mycobacterial immunity, to further our general understanding of mycobacterial pathogenesis. In particular, I will functionally characterize novel bi-allelic mutations I have recently discovered in IL12RB2 and IL23R, genes encoding the receptor chains that may pair with IL-12Rβ1 and are required for cellular responses to IL-12 and IL-23, respectively (Aim 1). These mutations are present in 2 kindreds with multiple MSMD-affected children. The receptors for IL-12 and IL-23 are considered components of either the canonical Th1 or Th17 T cell pathways, and are known to play critical roles in IFN-γ production, IL-17 production, or both. Therefore, I will carry out detailed mechanistic studies to understand the differential impacts of these novel monogenic lesions on the development and function of Th1 and Th17 cells (Aim 2). Finally, I will test the hypothesis that defects in IL-12 or IL-23 signaling cause MSMD in these patients, and seek to identify additional monogenic causes of MSMD in immunologically related genes in amongst >500 other MSMD patients (Aim 3). In the mentored K99 phase of the award, I will benefit from the unique learning environment and clinical resources of Dr. Jean-Laurent Casanova’s laboratory at the Rockefeller University (RU). Dr. Casanova’s lab is the world leader in monogenic dissection of infectious diseases, and interrogating susceptibility to mycobacteria is the lab’s longest-running project. While working in Dr. Casanova’s lab, I will enjoy unparalleled access to physicians and patients around the world, and the close counsel of many experienced colleagues. I will also gain experience with cutting-edge techniques including whole exome and whole genome sequencing, RNAseq, imaging flow cytometry, and multiplex cytokine arrays. A critical part of my career development plan is the scientific collaboration and career development advice offered by my Advisory Committee, consisting of Dr. Casanova, Dr. Luigi Notarangelo (NIAID/NIH), and Dr. Stuart Tangye (Garvan Institute, Australia), all of whom have pledged to support me in my proposed research and in the development of my independent career. In all, the proposed plan will fulfill my short-term scientific goal of elaborating the single-gene lesions responsible for mycobacterial susceptibility in children, and my long-term goal of attaining an independent role investigating the host genetic determinants of immunity to pathogens.