Human genetic determinants of mycobacterial immunity

项目总结/摘要 这个独立之路提案的主要目标是全面了解IL-12的作用 和IL-23信号在人类分枝杆菌免疫中的作用。分枝杆菌病的孟德尔易感性 （MSMD）是一种遗传性疾病，使儿童在感染弱毒力 包括卡介苗在内的分枝杆菌。虽然MSMD很罕见，但对这种疾病的研究有能力 阐明对毒性更强的结核分枝杆菌的免疫的基本免疫机制。在 在建议的研究计划中，我将使用从“自然实验”中收集的知识，即先天错误 的分枝杆菌免疫，以进一步我们对分枝杆菌的发病机制的一般理解。我特别 我将在功能上描述我最近在IL 12 RB 2和IL 23 R基因中发现的新的双等位基因突变， 编码可与IL-12 R β1配对并为细胞对IL-12应答所需的受体链， IL-23（目的1）。这些突变存在于2个有多个MSMD受影响的儿童的激酶中。 IL-12和IL-23的受体被认为是典型的Th 1或Th 17 T细胞的组分 在IFN-γ产生、IL-17产生或两者中起关键作用。所以我会 进行详细的机制研究，以了解这些新的单基因病变的不同影响 Th 1和Th 17细胞的发育和功能（目的2）。最后，我将检验以下假设： IL-12或IL-23信号传导导致这些患者的MSMD，并试图确定其他单基因原因， 在>500名其他MSMD患者中的免疫相关基因中的MSMD（目的3）。在K99的指导下 在获奖的第一阶段，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世界上单基因 传染病的解剖和对分枝杆菌的易感性的询问是实验室运行时间最长的 项目在卡萨诺瓦博士的实验室工作期间，我将享受无与伦比的接触医生和病人的机会。 世界，以及许多经验丰富的同事的密切建议。我也将获得经验， 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 多重细胞因子阵列。我的职业发展计划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建议，包括卡萨诺瓦博士，路易吉·诺塔朗杰洛博士 （NIAID/NIH）和Stuart Tangye博士（Garvan研究所，澳大利亚），他们都承诺支持我的研究。 在我的独立职业生涯的发展中。总的来说，提出的计划将实现我的目标。 短期科学目标是阐明导致分枝杆菌易感性的单基因病变， 儿童，我的长期目标是获得一个独立的作用，调查宿主的遗传决定因素， 对病原体的免疫力。

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.