Functional mapping of ex-regulatory T cell phenotypic diversity

项目总结/摘要 调节性T细胞和效应性T细胞具有不同的免疫学作用，有助于免疫稳态。 尽管效应T细胞具有保护宿主免受感染威胁和癌症的功能能力， 调节性T细胞（TCRs）本质上是抑制性的，并防止失控的效应T细胞介导的炎症 和自身免疫虽然这些是主要在胸腺选择过程中决定的不同细胞谱系，但它们是不同的。 现在了解到，这两个可以相互转换- T细胞可以在外周中呈现调节表型， 并且T细胞可能失去成为促炎性“非调节性”T细胞（exT细胞）的抑制能力。 因为exTlymphocyte可以颠覆传统的调节性T细胞来加剧炎症，并有助于 病理性自身免疫进展，有很大的兴趣，了解如何和在什么条件下， 外延发展。在我们实验室的初步工作中，我们发现单细胞RNA测序揭示了广泛的 来自小鼠肠、肺和淋巴结的原发性Tcl 4和外Tcl 4的表型异质性。我提议 使用单细胞基因组学方法来完全定义管理exTreg的基因调控程序 发展和两极分化的不稳定的Tibetan。在我的第一个目标中，我将深入分析单细胞 用于定义不同组织中exTreg表型异质性的RNA测序数据集 他们被隔离的微环境。对于我的第二个目标， 在高通量遗传扰动原代小鼠THBE中进行正向遗传筛选，以定位THBE基因。 维持Treg身份并使Treg不稳定的基因调控程序， 促炎细胞因子IL-4或IL-6。我的担保人亚历克斯·马森在遗传学方面有着广泛的专业知识， 免疫细胞功能和自身免疫，以及使用CRISPR-Cas9的原代T细胞的基因工程。我 共同发起人Chun（Jimmie）Ye是一位计算免疫学家和人类遗传学家，他的实验室使用 创新技术，如单细胞RNA测序，以分子方式定义免疫细胞的过程 发展和功能。此外，我还获得了来自著名的科学和职业导师 免疫学家Jeffrey Bluestone博士（自身免疫性炎症中的特征性外分泌）和Qizhi Tang博士 （研究免疫耐受和自身免疫）。与此同时，我正在完成一个纵向临床 与UCSF Benioff儿童医院主任Alice Chan博士一起担任儿科免疫学导师 儿科免疫失调诊所和儿科免疫疾病基因诊断专家。总的来说， 这项工作将进一步加深我们对exTlymphoma的发展及其在自身免疫性疾病中的作用的理解。 发病机制和进展。此外，这项奖学金支持将促进我在免疫学方面的培训， 计算生物学，支持我在我的道路上，通过结合MD-PhD培训走向职业生涯， 研究免疫紊乱遗传基础的儿科免疫学家。

PROJECT SUMMARY/ABSTRACT Regulatory and effector T cells serve distinct immunological roles that contribute to immune homeostasis. Whereas effector T cells have the functional capacity to protect the host from infectious threats and cancer, regulatory T cells (Tregs) are suppressive in nature and prevent runaway effector T cell-mediated inflammation and autoimmunity. Although these are distinct cell lineages determined primarily during thymic selection, it is now understood that the two can interconvert – T cells can assume regulatory phenotypes in the periphery, and Tregs may lose suppressive capacity to become pro-inflammatory “ex-regulatory” T cells (exTregs). Because exTregs can subvert traditional regulatory T cells to exacerbate inflammation and contribute to pathologic autoimmune progression, there is great interest in understanding how and under what conditions exTregs develop. In preliminary work from our lab, we found that single cell RNA sequencing reveals extensive phenotypic heterogeneity of primary Tregs and exTregs from mouse gut, lungs, and lymph nodes. I propose using single cell genomics approaches to fully define the gene regulatory programs that govern exTreg development and polarization from destabilized Tregs. In my first aim, I will deeply analyze the single cell RNA sequencing data set to define the heterogeneity of exTreg phenotypes in distinct tissue microenvironments from which they were isolated. For my second aim, I will leverage my sponsors’ expertise in high-throughput genetic perturbation of primary mouse Tregs to perform a forward genetic screen to map the gene regulatory programs that maintain Treg identity and destabilize Tregs to exTregs induced by pro-inflammatory cytokines IL-4 or IL-6. My sponsor Alex Marson has extensive expertise in the genetics of immune cell function and autoimmunity, and in genetic engineering of primary T cells using CRISPR-Cas9. My co-sponsor Chun (Jimmie) Ye is a computational immunologist and human geneticist whose laboratory uses innovative technologies like single cell RNA sequencing to molecularly define processes of immune cell development and function. Additionally, I have secured scientific and career mentorship from renowned immunologists Dr. Jeffrey Bluestone (characterized exTregs in autoimmune inflammation) and Dr. Qizhi Tang (studies immune tolerance and autoimmunity). Concurrently, I am completing a longitudinal clinical preceptorship in pediatric immunology with Dr. Alice Chan, director of the UCSF Benioff Children’s Hospital Pediatric Immune Dysregulation Clinic and expert in genetic diagnosis of pediatric immune disorders. Overall, the proposed work will further our understanding of exTregs development and their role in autoimmune pathogenesis and progression. Moreover, this fellowship support will foster my training in immunology and computational biology, supporting me on my path through combined MD-PhD training towards a career as an academic pediatric immunologist studying the genetic bases of immune disorders.