Understanding the role of Id2 in T cell differentiation and activation during GVHD

摘要 骨髓移植(BMT)可治愈多种血液系统恶性肿瘤，但移植物与 宿主病(GVHD)是供体细胞对宿主的免疫反应，仍然是最致命的风险 心理治疗。T淋巴细胞在宿主体内驱动炎症反应，可导致器官损伤和 破坏，甚至移植受者的死亡。虽然广泛的免疫抑制剂被用来预防 而治疗GVHD，确诊后仍会导致高达50%的死亡率。移植物抗宿主病有两种形式，急性 和慢性的，它们的发展时间和病理都不同。有必要更有针对性地 预防移植物抗宿主病的治疗，同时仍保持对病原体和 恶性复发。我们使用了我们的高度翻译相关的非人类灵长类(NHP)GVHD模型 目的：鉴定GVHD靶器官中T细胞上调的基因。通过转录转录途径分析，我们 已经发现DNA结合抑制物2(ID2)是许多基因激活的关键上游调节因子 移植物抗宿主病期间靶器官的破坏。在文献中，Id2参与调节T细胞分化，但 这是如何发生的，以及它如何影响移植物抗宿主病背景下T细胞的功能仍不清楚。这 该项目旨在了解T细胞中ID2缺失如何影响其分化为1型和17型(Th/c)的能力 驱动GVHD的T细胞和预防GVHD的调节性T细胞(Treg)。我们将使用 CRISPR/Cas9首次删除1型和17型T细胞中的ID2并在体外和GVHD中探索其功能 模特们。我们还将评估ID2编辑的T细胞在基线和 通过转录剪裁分析了解GVHD的背景。其次，我们将从Treg细胞中删除ID2以确定 Id2在维持其抑制活性中的作用。我们将评估ID2-Delteed Treg细胞的能力 为了控制移植物抗宿主病，特别是在慢性移植物抗宿主病中，Tregs在抑制正常T细胞中起重要作用。 从长远来看。靶向1型和17型T细胞的致病作用，同时避免T调节的保护作用 细胞在骨髓移植领域仍然是一个挑战，文献表明靶向ID2可能倾向于 宽容。这个项目将对更好地理解ID2在T细胞分化中的作用以及如何 靶向ID2可能为控制或治疗移植物抗宿主病提供了一种新的治疗策略。

Abstract Bone Marrow Transplantations (BMT) can be curative for a variety of hematologic malignancies but Graft Versus Host Disease (GVHD), an immune reaction of donor cells against the host, remains the deadliest risk of this therapy. T lymphocytes drive an inflammatory response in the host that can result in organ damage and destruction, and even death of the transplant recipient. While broad immunosuppressants are given to prevent and treat GVHD, it still results in the mortality of up to 50% after diagnosis. GVHD occurs in two forms, acute and chronic, that differ in their time to development as well as pathology. There is a need for more targeted therapies to prevent GVHD, while still maintaining the protective immune functions against pathogens and malignant relapse. We have used our highly translationally relevant non-human primate (NHP) model of GVHD to identify genes upregulated in T cells in target organs of GVHD. Through transcriptomic pathway analysis we have identified inhibitor of DNA-binding 2 (ID2) as a key upstream regulator of many genes activated in the destruction of target organs during GVHD. In the literature Id2 is involved in regulating T cell differentiation, but how this occurs and how it affects the functionality of T cells in the context of GVHD remains unknown. This project aims to understand how ID2 deletion in T cells affects their ability to differentiate into Type 1 and 17 (Th/c 1 and Th/c 17) T cells which drive GVHD and regulatory T cells (Treg) which prevent GVHD. We will use CRISPR/Cas9 to first delete ID2 in type 1 and 17 T cells and explore their functionality in vitro and in GVHD models. We will also assess how the regulatory landscape of ID2-edited T cells changes at baseline and in the context of GVHD through transcriptomic profiling. Second, we will delete ID2 from Treg cells to determine the effects of id2 in maintaining their suppressive activity. And we will evaluate the ability of ID2-delteted Treg cells to control GVHD, especially in cGVHD where Tregs play a vital role in suppressing conventional T cells in the long term. Targeting the pathogenicity of type 1 and 17 T cells while sparing the protective effects of T regulatory cells remains a challenge in the BMT field and the literature suggests that targeting ID2 may tip both towards tolerance. This project will be important in understanding better the role of ID2 in T cell differentiation and how targeting ID2 may represent a novel therapeutic strategy for controlling or treating GVHD.