Mechanisms of regulatory T cell processes by IL-2

IL-2 调节 T 细胞过程的机制

• 批准号: 10490273
• 负责人: Acacia Nicole Crouch
• 金额: $ 5.18万
• 依托单位: UNIVERSITY OF MIAMI SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10490273
• 关键词: ATAC-seq; Ablation; Acute; Address; Adoptive Transfer; Adrenal Cortex Hormones; Affect; American; Animals; Apoptosis; Autoimmune Diseases; Autoimmunity; Bioinformatics; Biological Assay; Biology; Bypass; Cause of Death; Cell physiology; Complex; Complication; Data; Defect; Development; Disease; Dose; Drops; Epigenetic Process; Evaluation; Exhibits; Fellowship; Flow Cytometry; Genetic Risk; Genetic Transcription; Heterogeneity; Homeostasis; Human; IL2RA gene; Immune; Incidence; Inflammatory; Insulin-Dependent Diabetes Mellitus; Interleukin 2 Receptor; Interleukin-2; Investigation; Knock-out; Knockout Mice; Knowledge; Lead; Lupus; Malignant Neoplasms; Manipulative Therapies; Measures; Mediating; Memory; Metabolism; Modeling; Molecular; Mus; Mutation; Pathway interactions; Peripheral; Play; Population; Positioning Attribute; Prevention; Process; Receptor Signaling; Recombinant Interleukin-2; Regulatory T-Lymphocyte; Research; Role; Self Tolerance; Signal Transduction; Techniques; Therapeutic; Training; Transcription Process; Woman; autoreactive T cell; cytokine; experience; improved; in vitro Assay; in vivo; middle age; mouse model; mutant; overtreatment; peripheral tolerance; side effect; tool; transcriptome sequencing

Autoimmune disease is one of the leading causes of death for young and middle-aged women in the US. These diseases are mediated by autoreactive T cells, which are naturally contained by T regulatory cells (Tregs). Non-functional mutations in the IL-2 receptor (IL-2R) lead to autoimmunity due to deficiencies in Treg survival and function. This finding is corroborated by in vivo mouse models, leading to the widespread conclusion that IL-2 is important for Treg survival. However, recent data indicates this is oversimplified. Our lab studies a mouse model in which the alpha chain (CD25) of the IL-2R on peripheral Tregs is conditionally ablated. CD25 knockout Tregs demonstrate a disproportionate loss of central Tregs, which are long-lived circulating Tregs that give rise to effectors to maintain self-tolerance. Bulk knockout Tregs also display disruptions in critical Treg processes such as proliferation, survival, metabolism and suppressive function. Additionally, there is an absence of highly suppressive terminally differentiated Tregs in IL-2R mutant animals, indicating IL-2 may regulate the development of highly suppressive Tregs required for maintain peripheral tolerance. Given these findings, IL-2 signaling controls processes that are essential for the prevention of autoimmunity and understanding this role can provide therapeutic avenues to manipulate this population. Aim 1 will determine the effects of IL-2 on survival, proliferation, function and metabolism in Treg subsets. Using flow cytometry and in vitro assays, proliferation, apoptosis, survival, function and metabolism will be examined in Tregs subsets with and without IL-2 signaling. This aim will inform the extent to which IL-2 regulates cellular processes in Tregs subsets to further define the way by which IL-2 maintains Treg populations. Aim 2 will evaluate the role of IL-2 in Treg differentiation and heterogeneity. Due to defects observed in the differentiation of Tregs in the setting of IL-2, this aim will use adoptively transferred Treg subsets with and without IL-2 signaling to determine how IL-2 regulates development of eTregs from cTregs. Aim 3 will define the distinct IL-2 dependent transcriptional and epigenetic processes in cTregs versus eTregs. This aim will harness high throughput techniques such as RNAseq and ATAC-seq to correlate epigenetic and transcriptional changes as a consequence of IL-2 signaling, or lack thereof, in Treg subsets. By defining IL-2-dependent pathways and cellular processes that differ between them, these findings will be integrated with those in aim 1 to provide a clear role for IL-2 in regulating the cellular processes in each Treg subset. The main objective of this fellowship research is to define IL-2 driven processes coordinating Treg survival and homeostasis that are acutely relevant for understanding therapeutic applications of this cytokine and broadly relevant for understanding autoimmune disease.

自身免疫性疾病是美国年轻和中年妇女的主要死亡原因之一。 这些疾病是由自动反应性T细胞介导的，T细胞自然包含。 （Tregs）。 IL-2受体（IL-2R）中的非功能突变导致自身免疫性由于Treg缺乏 生存和功能。该发现通过体内鼠标模型证实了这一发现，导致广泛 结论IL-2对于Treg生存很重要。但是，最近的数据表明这太过了。我们的实验室 研究一个小鼠模型，其中IL-2R的α链（CD25）在周围Tregs上是有条件的 消融。 CD25敲除Tregs表明中央Treg的损失不成比例，这是长寿的 循环treg会引起效应子，以保持自我耐受。散装淘汰赛也显示 关键的Treg过程中的破坏，例如增殖，生存，代谢和抑制作用。 此外，在IL-2R突变动物中没有高度抑制的终末分化的Treg， 表明IL-2可以调节维持周围所需的高度抑制性Treg的发展 宽容。鉴于这些发现，IL-2信令控制过程对于预防的过程至关重要 自身免疫性和理解这一角色可以提供治疗途径来操纵这一人群。目的 1将确定IL-2对Treg子集生存，增殖，功能和代谢的影响。 使用流式细胞术和体外测定，增殖，凋亡，生存，功能和代谢将是 在带有和没有IL-2信号传导的Tregs子集中检查。这个目标将告知IL-2的程度 调节Tregs子集中的细胞过程，以进一步定义IL-2维持Treg的方式 人群。 AIM 2将评估IL-2在Treg分化和异质性中的作用。由于缺陷 在IL-2的环境中，在Tregs的区分中观察到，此目标将采用转移的Treg 带有和没有IL-2信号传导的子集确定IL-2如何调节CTREG的ETREG的发展。 AIM 3将定义Ctregs中不同的IL-2依赖性转录和表观遗传过程 Etregs。此目标将利用高吞吐量技术，例如RNASEQ和ATAC-SEQ相关 Treg子集中IL-2信号传导或缺乏表观遗传和转录变化。经过 定义IL-2依赖性途径和它们之间不同的细胞过程，这些发现将是 与AIM 1中的那些集成在一起，以在调节每个Treg的细胞过程中为IL-2提供明确的作用 子集。这项研究金研究的主要目的是定义IL-2驱动的过程协调Treg 与理解该细胞因子的治疗应用非常相关的生存和稳态 与理解自身免疫性疾病的广泛相关。