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信号已被证明是激活T细胞功能的关键调节剂。 PD-L1对PD-1的参与降低了效应的T细胞增殖和细胞因子的产生，并诱导T细胞周期停滞和凋亡或促进IL-10产生的调节性T细胞反应。然而，几乎几乎没有知道PDL1相关信号在T细胞对（3细胞抗原和T1D过程的反应中）的作用。最近的研究表明，NOD小鼠在APC上的PDL1表达不足，胰岛特异性转基因的PDL1的PDL1显着降低了胰岛素和预防糖尿病的快速降低和预防NOD糖尿病的固定剂，PD的固定剂量是nod-blocky inod-blocky inod-blocky inod-blocked inod-deartly inod-pd the nod-deartly and block the nod-deftry nod-pd。我们的初步研究中的糖尿病发作表明，激动品PDL1-LG抑制了对P细胞抗原的自发性TH1反应，糖尿病性T细胞在体外的治疗，并用PDL1-LG进行PDL1-LG进行了延伸。腺体相关病毒（RAAV）-PDLI-LG，但没有RAAV-LG，在糖尿病NOD小鼠中长时间的胰岛移植生存率，并用RAAV-PDL1-LG诱导PDL1-LG的稳定表达PDL1-LG的6周大NOD小鼠，抑制了PDL1-LG的稳定表达，抑制了抑制疾病的进展，与自发性T细胞自动抑制相关。因此，我们假设用RAAV-PDL1-LG治疗诱导PDL1-LG稳定表达可能会使效应T细胞失活和/或诱导调节性T细胞反应，从而抑制NOD小鼠的晚期T1D和反向T1D的疾病进展。我们选择使用AAV作为载体将PDL1-LG或对照Ig基因输送到点头小鼠，因为AAV是一种非致病病毒，几乎没有免疫原性，使其对临床应用有吸引力。 AAV可以有效感染广泛的细胞，从而导致靶基因的长期表达。在该提案中，我们将研究自身免疫过程后期用Raav-PDL1-LG治疗对NOD小鼠中新糖尿病的逆转以及逆转NOD小鼠的新糖尿病的作用，并确定与PDL1相关信号在NOD小鼠中T细胞自动免疫性过程中与PDL1相关信号的作用。我们的研究将解决有关PD-1/PDL1信号在自身免疫T细胞反应中的调节功能的基本问题。我们的发现可能为人类T1D抑制/逆转的新型免疫疗法提供了基础。外行摘要：我们将集中于检查工程抑制剂对T细胞自身免疫性对小鼠模型中1型糖尿病的抑制/逆转的治疗作用。我们的发现可能为T1D患者设计特定免疫疗法的设计提供了基础。