PDL1-based rAAV-mediated gene therapy for mouse T1D

基于 PDL1 的 rAAV 介导的小鼠 T1D 基因治疗

• 批准号: 7094761
• 负责人: JIDE TIAN
• 金额: $ 15.45万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-05-01 至 2008-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7094761
• 关键词: CD28 molecule; NOD mouse; T lymphocyte; adeno associated virus group; apoptosis; autoimmunity; biological signal transduction; cell cycle; cell growth regulation; cell mediated lymphocytolysis test; diabetes mellitus; diabetes mellitus therapy; disease /disorder onset; enzyme linked immunosorbent assay; flow cytometry; gene delivery system; gene therapy; immunoglobulin G; immunotherapy; ligands; nonhuman therapy evaluation; recombinant virus; stimulant /agonist; transfection /expression vector

DESCRIPTION (provided by applicant): The PD-1/PDL-1 signaling has been shown to be a crucial regulator for activated T cell function. Engagement of the PD-1 by PD-L1 down-regulates effector T cell proliferation and cytokine production, and induces T cell cycle arrest and apoptosis or promotes IL-10-producing regulatory T cell responses. However, little is known the role of PDL1 -related signaling in T cell responses to (3-cell antigens and the T1D process. Recent studies have shown that NOD mice are deficient in expression of PDL1 on APCs, islet-specific transgenic expression of PDL1 significantly reduced insulitis and prevented diabetes in NOD mice, and treatment with blockade for PD-1/PDL1 signaling rapidly precipitated diabetes onset in young NOD mice. Our preliminary studies showed that agonistic PDL1-lg inhibited spontaneous Th1 response to p-cell antigens, arrested diabetogenic T cell cycling in vitro, and treatment with PDL1-lg retarded the onset of adoptively transferred T1D in vivo. In addition, transplantation with syngenic islets transduced by recombinant adeno-associated virus (rAAV)-PDLI-lg, but not rAAV-lg, prolonged islet-graft survival in diabetic NOD mice and treatment of 6 weeks old NOD mice with rAAV-PDL1-lg induced stable expression of PDL1-lg, inhibited disease progression, associated with inhibition of spontaneous T cell autoimmunity. Hence, we hypothesize that treatment with rAAV-PDL1-lg to induce PDL1-lg stable expression may inactivate effector T cells and/or induce regulatory T cell responses, inhibiting disease progression at advanced pre-T1D and reverse T1D in NOD mice. We choose to use AAV as the vector to deliver PDL1-lg or control Ig genes to NOD mice because AAV is a non-pathogenic virus and has little immunogenicity, making it attractive for clinical applications. AAV can effectively infect broad types of cells, leading to a long-term expression of target genes. In this proposal, we will examine the effect of treatment with rAAV- PDL1-lg at the late stage of autoimmune process on disease progression and on reversal of newly diabetes in NOD mice and determine the mechanism(s) underlying the action of PDL1-related signaling on the process of T cell autoimmunity in NOD mice. Our studies will address fundamental questions concerning the regulatory function of PD-1/PDL1 signaling on autoimmune T cell responses. Our findings may provide the basis for novel immunotherapies for the inhibition/reversal of T1D in human. Layperson's abstract: We will center on examining the therapeutic effect of an engineered inhibitor for T cell autoimmunity on inhibition/reversal of type 1 diabetes in mouse model. Our findings may provide a basis for the design of specific immunotherapies for T1D patients.

描述（由申请人提供）：PD-1/PDL-1信号传导已被证明是激活T细胞功能的关键调节因子。PD-L1参与PD-1可下调效应T细胞增殖和细胞因子的产生，诱导T细胞周期阻滞和凋亡或促进il -10产生的调节性T细胞反应。然而，PDL1相关信号在T细胞对3细胞抗原的反应和T1D过程中的作用知之甚少。最近的研究表明NOD小鼠在apc上缺乏PDL1的表达，胰岛特异性转基因PDL1的表达可显著降低NOD小鼠的胰岛素炎并预防糖尿病，阻断PD-1/PDL1信号通路可迅速促进年轻NOD小鼠的糖尿病发病。我们的初步研究表明，激动性PDL1-lg抑制了Th1对p细胞抗原的自发反应，在体外阻止了糖尿病源性T细胞的循环，并且PDL1-lg治疗延缓了体内过继转移T1D的发生。此外，用重组腺相关病毒(rAAV)-PDLI-lg而非rAAV-lg转导的同源胰岛移植，延长了糖尿病NOD小鼠的胰岛移植存活时间，并用rAAV-PDL1-lg治疗6周龄NOD小鼠，诱导了PDL1-lg的稳定表达，抑制了疾病进展，并与自发性T细胞自身免疫的抑制有关。因此，我们假设用rAAV-PDL1-lg诱导PDL1-lg稳定表达可能会灭活效应T细胞和/或诱导调节性T细胞反应，抑制NOD小鼠晚期T1D前的疾病进展并逆转T1D。我们选择AAV作为载体向NOD小鼠传递PDL1-lg或控制Ig基因，因为AAV是一种非致病性病毒，免疫原性小，具有临床应用的吸引力。AAV可以有效感染多种类型的细胞，导致靶基因的长期表达。在本研究中，我们将研究自身免疫过程晚期rAAV- PDL1-lg治疗对NOD小鼠疾病进展和新发糖尿病逆转的影响，并确定pdl1相关信号在NOD小鼠T细胞自身免疫过程中的作用机制。我们的研究将解决PD-1/PDL1信号对自身免疫T细胞反应的调节功能的基本问题。我们的发现可能为抑制/逆转人类T1D的新免疫疗法提供基础。外行人摘要：我们将集中研究T细胞自身免疫工程抑制剂在小鼠模型中对1型糖尿病的抑制/逆转的治疗效果。我们的发现可能为设计针对T1D患者的特异性免疫疗法提供基础。