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

• 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
• 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

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.

项目摘要/摘要 该项目的目的是增强我们对IDG符合条件的激酶Dyrk1b，dyrk2， dyrk3和dyrk4。我们的初步数据牵涉到至少这些激酶中的一种 抗炎tregs。非常需要进行Treg调节疗法，尤其是在自身免疫性方面。我们建立了一个 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，但不调节Treg，分化。现在，我们建议使用此管道来识别 treg调节的dyrk。这将为IDG符合条件的基因的新型免疫细胞表型阐明，发展 经过验证的工具研究原始细胞中的IDG-DYRK，为新型Treg增强药物的开发提供信息 突出显示患者子集用于精确治疗。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，以治疗自身免疫性。此应用程序中的总体目标是 （i）开发遗传工具来确定哪些dyrk家族成员调节treg差异，以及（ii）询问 DYRK缺陷Tregs的功能和机械特征。中心假设是一个身份不明的dyrk 家庭成员抑制Treg分化。该项目的理由是识别Treg调节的Dyrk 家庭成员将提供一个强大的科学框架，以阐明我们对已研究的dyrk的理解 作为自身免疫性病理生物学的可毒品调节剂，并建立一条管道以阐明其他IDG资格 基因。中心假设将以两个特定的目的进行测试：1）定义如何过表达IDG-Dyrks 影响Treg的分化，2）定义IDG DYRK抑制剂和IDG-DYRK的敲除如何影响Treg 分化。第一个目标将询问主要鼠类中每个dyrk家庭成员的过表达 人CD4+ T细胞会影响Treg分化。第二个目标将机械地询问如何淘汰 原代鼠和人CD4+ T细胞中的每个DYRK家族成员都会影响Treg的分化和功能。 这些研究利用了我们发表的实验管道，该管道量化了遗传或化学的方式 扰动会影响T细胞分化。该提案的关键创新特征包括研究免疫 IDG-Dyrks的功能和原代免疫细胞的使用。拟议的研究很重要，因为它是 预计（i）揭示了一个新的细胞表型，用于IDG感兴趣的研究基因，（ii）确定一种新颖的 自身免疫性病理生物学的可药物调节剂，因此指导未来的机械和治疗研究，（iii） 开发有特征的工具来操纵在其他原始细胞和生物学环境中的研究dyrks，以及 （iv）建立可扩展的管道，该管道可用于快速询问IDG的所有感兴趣的基因，以实现影响 区分TH17，TH1和TH2在内的Treg以及其他谱系。最终，这有潜力 广泛阐明IDG基因的免疫功能并提高自身免疫性的精度治疗 在一般人群中。