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.

项目总结/摘要 该项目的目标是提高我们对IDG合格激酶DYRK 1B，DYRK 2， DYRK 3和DYRK 4。我们的初步数据暗示这些激酶中至少有一种在调节细胞分化中起作用。 抗炎的Tibetan。Treg调节疗法是非常需要的，特别是在自身免疫中。我们建立了一个 发表的管道，以量化遗传和化学扰动如何影响Treg分化，这表明 DYRK 1A调节Th 17而不是Treg分化。我们现在建议利用这条管道来确定 调节Treg的DYRK。这将阐明一种新的免疫细胞表型的IDG合格基因，开发 研究原代细胞中IDG-DYRKs的有效工具，为新型Treg增强药物的开发提供信息， 突出显示患者子集以进行精确治疗。我们的长期目标是帮助开发更好的疗法，包括 特异性DYRK家族成员的抑制剂，以治疗自身免疫。本申请的总体目标是 （i）开发遗传工具以鉴定哪个DYRK家族成员调节Treg分化，和（ii）询问 DYRK缺陷型THEORY的功能和机制特征。核心假设是一个未被确认的DYRK 家族成员抑制Treg分化。该项目的基本原理是，确定调节Treg的DYRK 家庭成员将提供一个强有力的科学框架，以阐明我们对未充分研究的DYRK的理解 作为自身免疫病理学的可药物调节剂，并建立一个管道，以照亮其他符合IDG标准的 基因.中心假设将在两个具体目标中进行检验：1）定义IDG-DYRKs的过表达如何影响IDG-DYRKs的表达。 影响Treg分化，以及2）定义IDG-DYRK抑制剂和IDG-DYRK敲除如何影响Treg 分化第一个目标是研究DYRK家族成员在原代小鼠中的过表达是如何发生的。 而人CD 4 + T细胞影响Treg分化。第二个目标将机械地询问如何击倒 原代鼠和人CD 4 + T细胞中每个DYRK家族成员的表达影响Treg分化和功能。 这些研究利用了我们已发表的实验管道，该管道量化了遗传或化学 干扰影响T细胞分化。该提案的主要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。这项研究之所以重要，是因为 预期（i）揭示IDG感兴趣的未充分研究的基因的新细胞表型，（ii）鉴定新的 自身免疫病理生物学的可药用调节剂，从而指导未来的机制和治疗研究，（iii） 开发特征化的工具，在其他原代细胞和生物学背景下操纵研究不足的DYRKs， (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Th 17以及其他谱系，包括Th 17、Th 1和Th 2。最终，这有可能 广泛阐明IDG基因的免疫功能，推进人类自身免疫的精准治疗 在普通人群中的比例。