Illuminating a novel role of understudied DYRKs in anti-inflammatory T cell differentiation

阐明正在研究的 DYRK 在抗炎 T 细胞分化中的新作用

基本信息

批准号：
10217878
负责人：
Bernard Khor
金额：
$ 17.41万
依托单位：
BENAROYA RESEARCH INST AT VIRGINIA MASON
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-05-01 至 2023-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10217878
关键词：
Affect Animal Model Anti-Inflammatory Agents Attenuated Autoimmune Autoimmune Diseases Autoimmunity Biological CD4 Positive T Lymphocytes CRISPR/Cas technology Cells Cellular biology Chemicals Chromosome 21 Data Development Disease Down Syndrome Enhancers Family member Foundations Funding Future General Population Genes Genetic Goals Harmine Human Immune Impairment Individual Inflammation Inflammatory Knock-out Knowledge Methods Mission Molecular Target Mus Patients Pharmaceutical Preparations Phenotype Phosphotransferases Pilot Projects Precision therapeutics Public Health Publishing Regulatory T-Lymphocyte Research Role Signal Transduction T cell differentiation T-Lymphocyte Testing Therapeutic Studies Transgenic Organisms Treg therapy United States National Institutes of Health Work base chemical genetics design disability genetic approach immune function inhibitor/antagonist innovation insight interest member novel novel therapeutics overexpression patient subsets personalized immunotherapy side effect small hairpin RNA small molecule success tool

项目摘要

Project Summary/Abstract The goal of this project is to enhance our understanding of the IDG-eligible kinases DYRK1B, DYRK2, DYRK3 and DYRK4. Our preliminary data implicate at least one of these kinases in regulating differentiation of anti-inflammatory Tregs. Treg-modulating therapies are greatly needed, particularly in autoimmunity. We built a published pipeline to quantitate how genetic and chemical perturbations impact Treg differentiation, this showed that DYRK1A regulates Th17, but not Treg, differentiation. We now propose to use this pipeline to identify the Treg-regulating DYRK. This will illuminate a novel immune cellular phenotype for an IDG-eligible gene, develop validated tools to study IDG-DYRKs in primary cells, inform development of novel Treg-enhancing drugs and highlight patient subsets for precision therapy. Our long-term goal is to help develop better therapies, including inhibitors of specific DYRK family members, to treat autoimmunity. The overall objectives in this application are to (i) develop genetic tools to identify which DYRK family member regulates Treg differentiation, and (ii) interrogate functional and mechanistic features of DYRK-deficient Tregs. The central hypothesis is that an unidentified DYRK family member inhibits Treg differentiation. The rationale for this project is that identifying the Treg-regulating DYRK family member will offer a strong scientific framework to illuminate our understanding of understudied DYRK(s) as druggable regulators of autoimmunity pathobiology and establish a pipeline to illuminate other IDG-eligible genes. The central hypothesis will be tested in two specific aims: 1) Define how overexpression of IDG-DYRKs affects Treg differentiation, and 2) Define how IDG DYRK inhibitors and knockout of IDG-DYRKs affects Treg differentiation. The first aim will interrogate how overexpressing each DYRK family member in primary murine and human CD4+ T cells affects Treg differentiation. The second aim will mechanistically interrogate how knockout of each DYRK family member in primary murine and human CD4+ T cells affects Treg differentiation and function. These studies leverage our published experimental pipeline that quantitates how genetic or chemical perturbations impact T cell differentiation. Key innovative features of this proposal include studying the immune function of IDG-DYRKs and the use of primary immune cells. The proposed research is significant because it is expected to (i) reveal a new cellular phenotype for an understudied gene of interest to IDG, (ii) identify a novel druggable regulator of autoimmune pathobiology, thus directing future mechanistic and therapeutic studies, (iii) develop characterized tools to manipulate understudied DYRKs in other primary cells and biologic contexts and (iv) establish a scalable pipeline that can be used to rapidly interrogate all genes of interest to IDG for effects on differentiation into Tregs as well as other lineages including Th17, Th1 and Th2. Ultimately, this has the potential to broadly illuminate immune function of IDG genes and advance precision therapy of autoimmunity for people in the general population.

项目概要/摘要该项目的目标是增强我们对 IDG 合格激酶 DYRK1B、DYRK2、 DYRK3 和 DYRK4。我们的初步数据表明这些激酶中至少有一种参与调节分化抗炎Tregs。非常需要 Treg 调节疗法，特别是在自身免疫方面。我们建造了一个已发表的管道来量化遗传和化学扰动如何影响 Treg 分化，这表明 DYRK1A 调节 Th17 分化，但不调节 Treg 分化。我们现在建议使用这个管道来识别 Treg 调节 DYRK。这将阐明 IDG 合格基因的新型免疫细胞表型，开发研究原代细胞中 IDG-DYRK 的经过验证的工具，为新型 Treg 增强药物的开发提供信息突出显示精准治疗的患者亚群。我们的长期目标是帮助开发更好的疗法，包括特定 DYRK 家族成员的抑制剂，用于治疗自身免疫性疾病。该应用程序的总体目标是 (i) 开发遗传工具来识别哪个 DYRK 家族成员调节 Treg 分化，以及 (ii) 探究 DYRK 缺陷的 Tregs 的功能和机制特征。中心假设是一个身份不明的 DYRK 家族成员抑制 Treg 分化。该项目的基本原理是确定 Treg 调节 DYRK 家庭成员将提供强大的科学框架来阐明我们对正在研究的 DYRK 的理解作为自身免疫病理学的药物调节剂，并建立一个管道来阐明其他符合 IDG 资格的药物基因。中心假设将在两个具体目标上进行检验：1) 定义 IDG-DYRK 的过度表达如何影响 Treg 分化，2) 定义 IDG DYRK 抑制剂和 IDG-DYRK 敲除如何影响 Treg 差异化。第一个目标将探究原代小鼠中每个 DYRK 家族成员如何过度表达人类 CD4+ T 细胞影响 Treg 分化。第二个目标将机械地询问如何淘汰赛原代小鼠和人类 CD4+ T 细胞中每个 DYRK 家族成员的变化都会影响 Treg 的分化和功能。这些研究利用我们发表的实验流程，定量遗传或化学扰动会影响 T 细胞分化。该提案的主要创新特点包括研究免疫 IDG-DYRK 的功能和初级免疫细胞的使用。拟议的研究意义重大，因为它预计将 (i) 揭示 IDG 感兴趣的尚未研究的基因的新细胞表型，(ii) 识别新的细胞表型自身免疫病理学的药物调节剂，从而指导未来的机制和治疗研究，(iii) 开发特征工具来操纵其他原代细胞和生物环境中未充分研究的 DYRK， (iv) 建立一个可扩展的管道，可用于快速询问 IDG 感兴趣的所有基因对分化为 Tregs 以及其他谱系，包括 Th17、Th1 和 Th2。最终，这有潜力广泛阐明IDG基因的免疫功能，推进人类自身免疫精准治疗在一般人群中。