Modulation of Teff/Treg cells by novel PKC-theta allosteric inhibitory strategies

通过新型 PKC-theta 变构抑制策略调节 Teff/Treg 细胞

• 批准号: 8771233
• 负责人: AMNON ALTMAN
• 金额: $ 26.55万
• 依托单位: LA JOLLA INSTITUTE FOR IMMUNOLOGY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-07-15 至 2016-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8771233
• 关键词: Address; Adoptive Transfer; Allergic; Antigens; Antiviral Agents; Asthma; Autoimmune Diseases; Autoimmunity; Biological; Blood; Bone Marrow; CD28 gene; CD4 Positive T Lymphocytes; Cell Culture Techniques; Cell Differentiation process; Cell physiology; Cells; Clinical; Complex; Data; Development; Disease; Disease model; Distal; Dominant-Negative Mutation; Drug Targeting; Effector Cell; Enzymes; Family member; Goals; Graft Rejection; Health; Human; Hypersensitivity; Immune response; Immune system; Immunity; Immunosuppression; In Vitro; Inflammation; Inflammatory Response; Laboratories; Lung Inflammation; Mediating; Modeling; Molecular; Mus; Ovalbumin; Play; Population; Proline; Protein Kinase C; Regulatory T-Lymphocyte; Reporter; Role; Signal Transduction; Stimulus; System; T cell response; T-Cell Activation; T-Lymphocyte; Th2 Cells; Therapeutic; Time; Transgenic Mice; Transgenic Organisms; Work; allograft rejection; base; immunological synapse; in vivo; inhibitor/antagonist; leukemia; mouse model; mutant; novel; pathogen; receptor; response; small molecule; tool

DESCRIPTION (provided by applicant): Protein kinase C theta (PKCθ) is a PKC family member most abundant in T cells, where it plays critical roles in effector T (Teff) cell activation PKCθ uniquely localizes to the center of the T cell immunological synapse (IS) via physical association with CD28, and this localization is required for downstream PKCθ-mediated signaling. In vivo analysis of PKCθ-deficient mice revealed a selective requirement of PKCθ in some (Th2- and Th17- mediated inflammation, allograft rejection and graft vs. host disease) but not other (Th1/CTL antiviral and graft vs. leukemia) immune responses. Intriguingly, PKCθ was recently found to be excluded from the IS of induced Treg cells (iTregs) and to inhibit their suppressive function, including in humans. Our preliminary studies show that strategies, which disrupt the PKCθ-CD28 interaction and displace PKCθ from the IS, including novel, previously uncharacterized PKCθ allosteric inhibitors (AI), enhance the in vitro differentiation and suppressive function of mouse iTregs. Our new preliminary data suggest that the same strategies have similar effects on human T cells. These findings point to the high promise of PKCθ as a selective drug target for immunosuppression of harmful T cell-mediated autoimmunity, inflammation, and GvHD, where inhibitory strategies are predicted to act synergistically to inhibit pathogenic T cells and promote Treg function, without interfering with beneficial antiviral protective immunity. Here, we will investigate in detail the potential therapeutic value of blocking the PKCθ-CD28 interaction in a mouse model of Th2 cell-mediated allergic lung inflammation, and the selective and seemingly opposite roles of PKCθ in the in vitro differentiation and function of human Teffs and Tregs, respectively. These studies will address the working hypothesis that blockade of the PKCθ-CD28 interaction inhibits the differentiation and function of pathogenic T cells and promotes the function of Tregs in both mice and humans, thereby achieving a beneficial synergetic effect against harmful T cell- mediated inflammatory responses. In Aim 1, we will employ a dominant negative PKCθ mutant and novel PKCθ AI to study the resulting effects on the development and manifestation of Th2-mediated allergic lung inflammation in intact mice, bone marrow chimeric mice, or by using adoptive T cell transfers, including the use of TCR-transgenic and FoxP3eGFP reporter mice. In Aim 2, we will use, for the first time, the same blocking strategies of Aim 1 to disrupt the TCR/CD28-induced PKCθ-CD28 interaction in human CD4+ T cells, and evaluate their effect on the in vitro differentiation and function of human blood-derived Teff and Treg cells. This project will extend our understanding of the importance of the PKCθ-CD28 interaction, which was discovered in our laboratory, and its biological relevance for the differentiation and function of both Teff and Treg cells, importantly extending these findings to human T cells. It will also represent the first detailed analysis of novel PKCθ AI on T cell responses, and may pave the way to the development of new, highly selective therapeutic strategies to treat T cell-mediated allergies, autoimmune, diseases, and GvHD.

描述（由应用提供）：蛋白激酶C theta（PKCθ）是T细胞中最丰富的PKC家族成员，它在效应T（Teff）细胞活化中起着关键作用PKCθ独特地定位于与CD28相关的T细胞免疫突触（IS）的中心，并且需要用于CD28的局部化，并且需要用于下层pkccipied pkCCCCCCCCCCCCCCCCCCC CCCC CC CC CC串联。对PKCθ缺陷小鼠的体内分析显示，在某些（Th2和Th17-介导的注射，同种异体移植排斥和移植物与宿主疾病）中，PKCθ的选择性要求，但没有其他（TH1/CTL抗病毒和Graft和Graft vs.Lerukemia）。有趣的是，最近发现PKCθ被排除在诱导的Treg细胞（ITREGS）之外，并抑制其抑制功能，包括人类。我们的初步研究表明，破坏PKCθ-CD28相互作用并从IS中取代PKCθ的策略，包括新颖的，以前未表征的PKCθ变构抑制剂（AI），增强了小鼠ITREG的体外分化和抑制功能。我们的新初步数据表明，相同的策略对人类T细胞具有相似的影响。这些发现表明，PKCθ是对有害T细胞介导的自身免疫，感染和GVHD进行免疫抑制的选择性药物靶标的，其中预计抑制性策略可以协同作用以抑制病原T细胞并促进Treg功能，而无需促进抗病毒性抗病毒性抗病毒性免疫性。在这里，我们将详细研究在Th2细胞介导的过敏性肺部感染的小鼠模型中阻断PKCθ-CD28相互作用的潜在治疗值，以及分别在人类Teffs和Tregs的体外分化和功能中，PKCθ的选择性和看似相反的作用。这些研究将解决以下假设：阻断PKCθ-CD28相互作用抑制了致病性T细胞的分化和功能，并促进了小鼠和人类中Treg的功能，从而实现了对有害T细胞介导的炎症反应的有益协同作用。在AIM 1中，我们将采用主要的负PKCθ突变体和新颖的PKCθAI来研究完整小鼠Th2介导的过敏性肺部炎症，骨髓嵌合小鼠的发育和表现的影响，或者通过使用适应性T细胞转移器，包括使用TCR- TCR-TRANSGENIC和FOXPC PHOP PHOP3 RECRORTER MICE。在AIM 2中，我们将首次使用AIM 1的相同阻断策略来破坏人类CD4+ T细胞中TCR/CD28诱导的PKCθ-CD28相互作用，并评估其对人类血液衍生的Teff和Treg细胞的体外分化和功能的影响。该项目将扩展我们对PKCθ-CD28相互作用的重要性的理解，该相互作用是在我们的实验室中发现的，其生物学相关性与Teff和Treg细胞的分化和功能相关性，重要的是将这些发现扩展到人T细胞。它还将代表对T细胞反应的新型PKCθAI进行的首次详细分析，并可能为开发新的，高度选择性的治疗策略铺平了道路，以治疗T细胞介导的过敏，自身免疫，疾病和GVHD。