Defining factors that control human innate lymphoid cell development and function

先天性淋巴样细胞(ILCs)是一个淋巴细胞家族，在协调免疫反应和调节组织动态平衡方面起着关键作用。虽然炎症性ILC在宿主防御和抗肿瘤免疫中发挥重要作用，但我们和其他人已经确定了抑制免疫反应或招募其他抑制性细胞的调节性ILC群体。动物研究已经提供了控制ILC身份和功能的转录网络的路线图，但对人类ILC的研究相对较少。研究人类ILC的发育、调节和可塑性是非常必要的，以了解指导人类ILC发育的途径，并确定微环境因素如何影响其功能。这项研究集中在描述控制人类ILC发展和功能的分子机制，特别是调节ILC。目标1：确定驱动ILCreg发育和功能的关键途径。我们的目标是利用先进的基因组学平台确定参与调节ILC发育和功能的途径和转录因子。使用已建立的分离和扩增体内来源的人ILCs的方法，将诱导的调节性ILCs与典型的ILCs进行对比。然后将探索与免疫抑制功能和组织修复活动相关的途径和分子的相关性。目的2：检测诱导的调节性ILC和规范的人类ILC亚群的可塑性。不同的细胞因子和微环境因素如何影响健康个体的循环和组织驻留的人类ILC尚未确定。我们的目标是通过分离ILCs，使它们暴露于各种细胞因子，并利用先进的基因组和免疫学平台在表观遗传、转录和蛋白质水平上对它们进行分析，来确定人ILCs在血液和健康人体组织(肾脏、胰腺、脾)中的稳定性或可塑性。还将检测诱导的调节性ILC在细胞因子刺激下维持其表型或转化为其他ILC亚群的能力。目标3：评估转录因子在ILC发展和功能中的作用。一个关键的问题是，是否有一个谱系定义了调节性ILC的转录因子。对人体内来源的调节性ILC的RNAseq分析确定了感兴趣的转录因子，目的1旨在识别区分诱导的调节性ILC的转录因子。在这里，我们将利用慢病毒基因转移系统来探索几种转录因子在人类ILC发育和功能中的作用。将研究这些转录因子下游的基因表达网络和ILC功能，确定调节ILC相关转录因子对ILC发展和功能的影响。新颖性：我们对健康个体的ILCregs和人类ILC的研究将扩大我们对控制人类ILC发育和功能的分子途径的理解。

Innate lymphoid cells (ILCs) are a family of lymphocytes with pivotal roles in orchestrating immune responses and regulating tissue homeostasis. While inflammatory ILCs serve important functions in host defence and anti-tumour immunity, we and others have identified regulatory ILC populations that inhibit immune responses or recruit other suppressive cells. Animal studies have provided a roadmap of transcriptional networks that control ILC identity and function, yet studies of human ILCs are relatively scarce. Studies addressing human ILCs development, regulation and plasticity are greatly needed to understand pathways that direct human ILC development and determine how microenvironment factors influence their function. This research is centred on delineating molecular mechanisms that control human ILC development and function, with a particular focus on regulatory ILCs. Aim 1: Define key pathways that drive ILCreg development and function. We aim to identify pathways and transcription factors involved in regulatory ILC development and function using advanced genomics platforms. Induced regulatory ILCs will be contrasted with canonical ILCs using established methods to isolate and expand in vivo-derived human ILCs. Relevance of pathways and molecules associated with immunosuppressive functions and tissue repair activity will then be explored. Aim 2: Examine the plasticity of induced regulatory ILCs and canonical human ILC subsets. How different cytokines and microenvironment factors influence circulating and tissue-resident human ILCs from healthy individuals has not been determined. We aim to determine the stability or plasticity of human ILCs in blood and healthy human tissues (kidney, pancreas, spleen) by isolating ILCs, exposing them to various cytokines and profiling them at the epigenetic, transcriptional and protein level using advanced genomic and immunological platforms. Induced regulatory ILCs will also be examined for their ability to maintain their phenotype or convert to other ILC subsets with cytokine stimulations.  Aim 3: Assess the role of transcription factors in ILC development and function. A key question is whether there is a lineage defining transcription factor for regulatory ILCs. RNAseq analysis of human in vivo-derived regulatory ILCs identified transcription factors of interest, and Aim 1 aims to identify transcription factors that distinguish induced regulatory ILCs. Here, we will explore the role of several transcription factors in human ILC development and function using a lentiviral gene transfer system. Gene expression networks and ILC functions downstream of these transcription factors will be examined, the influence of regulatory ILC-associated transcription factors on ILC development and function determined. Novelty: Our studies of ILCregs and human ILCs from healthy individuals will expand our understanding of molecular pathways that control human ILC development and function.