Gene Therapeutic Approach for Tolerance Induction

耐受诱导的基因治疗方法

• 批准号: 7922278
• 负责人: David William Scott
• 金额: $ 6.68万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-17 至 2010-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7922278
• 关键词: Adenoviruses; Animal Model; Antibody Formation; Antigen-Presenting Cells; Antigens; Area; Autoantigens; Autoimmune Diseases; Autoimmune Process; Automobile Driving; B-Lymphocyte Subsets; B-Lymphocytes; Blocking Antibodies; Cell Communication; Cell Line; Cells; Chimeric Proteins; Clinic; Data; Epitopes; Experimental Autoimmune Encephalomyelitis; Future; Generations; Goals; Human; IL2RA gene; IgG1; Immune response; In Vitro; Interleukin-12; Kinetics; Knockout Mice; Knowledge; Label; Lead; Longevity; Lymphoid Tissue; Maintenance; Measures; Mediator of activation protein; Methods; Microscopy; Modeling; Multiple Sclerosis; Mus; Myelin Associated Glycoprotein; Myelin Basic Proteins; Pathway interactions; Patients; Peptides; Phenotype; Prevention; Process; Production; Protocols documentation; Recruitment Activity; Reporting; Research; Retroviral Vector; Role; Site; Stimulus; System; T-Cell Proliferation; T-Lymphocyte; TNFSF5 gene; Technology; Testing; Therapeutic Intervention; Tissues; Transgenic Mice; Transgenic Organisms; Translating; Translations; anti-IgM; base; chemokine; clinical efficacy; clinically relevant; congenic; cytokine; design; gene therapy; human disease; in vivo; insight; intravital microscopy; migration; novel; novel strategies; oligodendrocyte-myelin glycoprotein; peripheral blood; prevent; public health relevance; response; success; therapeutic gene; two-photon; vector

DESCRIPTION (provided by applicant): Modulation of immune responses against CNS antigens, such as myelin basic protein (MBP) and myelin oligodendrocyte glycoprotein (MOG), is a goal for therapeutic intervention in multiple sclerosis. The focus of our lab has been to develop novel approaches for the induction of antigen-specific tolerance that can be applied to the prevention or reversal of undesirable immune responses, e.g., in autoimmune diseases like MS. During the last decade, we created a platform technology that can be used for gene therapy to induce tolerance to multiple epitopes. In our model, LPS-activated B cells are transduced via retroviral vectors to express an Ig fusion protein with the targeted epitopes. Data in four experimental autoimmune models have demonstrated clinical efficacy in that expression of Ig fusion proteins of autoantigens by B cells can both prevent and reverse autoimmune responsiveness. Importantly, we found that CD25+, FoxP3+ regulatory T cells were required for both the induction and maintenance of tolerance in this system. We also reported that the mode of activation of the transduced B cells was a critical factor for the success of B cell-delivered gene therapy for tolerance, an observation that is important for potential translation. We hypothesize that different B cell activators lead to the production of distinct sets of chemokines and cytokines and/or the stimulation of tolerogenic (marginal zone) versus non-tolerogenic B cells. Our hypothesis is that transduced antigen-presenting B cells (APC), in particular marginal zone B cells, present the processed epitopes to effector T cells and/or Tregs to inhibit responsiveness via regulatory cytokines. Further, we suggest that the tolerogenic B cells directly recruit Tregs via CTLA-4 to induce tolerance. In order to test these hypotheses and to translate our efforts to the clinic for potential future treatment of MS, we will focus in this renewal on the following three specific aims. (1) Based on the observation that LPS- and CpG-activated B cells produce different cytokines and chemokines, using knockout mice, we will define the roles of specific soluble mediators generated under these and other activation conditions. We will also analyze the lifespan and phenotype of transduced B cells, as well as their migration to lymphoid tissues and the CNS. We will test the hypothesis that anti-CD20 treatments leads to partial depletion of B cells, sparing the tolerogenic MZ cells. (2) We will determine the migration and fate of the TCR transgenic T cells in B cell delivered gene therapy, and will analyze the kinetics of their interaction with tolerogenic or non-tolerogenic B cells using two photon microscopy. (3) In our final aim, we will use "humanized" systems to begin to translate these studies to human disease by analyzing responses of HLA DR2 transgenic mice and of human T cells in vitro, including the activation of Tregs by gene therapy. We will also examine the efficacy of non-integrating vectors (e.g., gutless adenovirus) for B-cell delivered gene therapy in both the later system and in EAE models. These studies will provide proof of principle with human T cells in a clinically relevant model, will establish the mechanisms of B cell-delivered gene therapy for tolerance, and will move this project forward to translation in the clinic as a potential therapy for multiple sclerosis. PUBLIC HEALTH RELEVANCE T cells from multiple sclerosis patients have been shown to respond to a variety of CNS antigens, such as myelin oligodendrocyte glycoprotein (MOG), myelin basic protein (MBP) and phospholipoprotein (PLP); hence, modulation of these immune responses is a goal for therapeutic intervention in MS. The focus of our lab has been to develop a novel B-cell delivered gene therapy method for the induction of tolerance, particularly in autoimmune diseases like MS. This platform has been successfully applied in murine models for MS. Herein, we will investigate the mechanism of B-cell delivered tolerance in animal models, and then we will begin to translate these studies to the clinic using T cells from MS patients exposed to B cells transduced with MBP-Ig constructs in vitro. These studies will establish a novel approach for tolerance with human T cells in a clinically relevant model, and will continue the translation of this project forward to the clinic as a potential therapy for multiple sclerosis.

描述（由申请人提供）：针对CNS抗原的免疫反应的调节，例如髓磷脂碱性蛋白（MBP）和髓磷脂少突胶质细胞糖蛋白（MOG），是多发性硬化症中治疗干预的目标。我们实验室的重点是开发新的方法来诱导抗原特异性耐受性，这些耐受性可以应用于预防或逆转不良免疫反应，例如在MS等自身免疫性疾病中。在过去的十年中，我们创建了一种平台技术，该技术可用于基因治疗，以诱导多个表位的耐受性。在我们的模型中，通过逆转录病毒载体转导LPS激活的B细胞，以表达与靶向表位的Ig融合蛋白。四个实验自身免疫模型中的数据证明了B细胞对自身抗原的Ig融合蛋白的表达可以预防和反向自身免疫反应性。重要的是，我们发现CD25+ FOXP3+调节T细胞是该系统中耐受性的诱导和维持所必需的。我们还报道说，转导B细胞的激活方式是B细胞分配基因治疗成功耐受性的关键因素，这一观察结果对于潜在的翻译很重要。我们假设不同的B细胞活化剂导致产生不同的趋化因子和细胞因子和/或刺激耐受性（边缘区）与非胆红性B细胞的刺激。我们的假设是转导抗原呈递B细胞（APC），尤其是边缘区B细胞，将处理的表位呈现到效应T细胞和/或Tregs通过调节细胞因子抑制反应性。此外，我们建议耐受性B细胞通过CTLA-4直接募集Tregs以诱导耐受性。为了检验这些假设，并将我们的努力转化为诊所以寻求MS的未来治疗方法，我们将重点关注以下三个特定目标。 （1）基于LPS和CPG激活的B细胞产生不同的细胞因子和趋化因子的观察结果，我们将使用基因敲除小鼠定义在这些和其他激活条件下产生的特定可溶性介质的作用。我们还将分析转导B细胞的寿命和表型，以及它们迁移到淋巴组织和中枢神经系统的迁移。我们将检验以下假设：抗CD20治疗导致B细胞部分耗竭，从而避免耐受性MZ细胞。 （2）我们将在B细胞递送的基因疗法中确定TCR转基因T细胞的迁移和命运，并将使用两个光子显微镜分析它们与耐受性或非胆红性B细胞相互作用的动力学。 （3）在我们的最终目标中，我们将使用“人性化”系统开始通过分析HLA DR2转基因小鼠和人类T细胞的体外反应，将这些研究转化为人类疾病，包括通过基因治疗激活TREG。我们还将研究非整合载体（例如肠道腺病毒）在后来的系统和EAE模型中均提供基因治疗的疗效。这些研究将在临床上相关的模型中提供人类T细胞的原理证明，将建立B细胞分配基因疗法的耐受性机制，并将该项目推动该项目在诊所转化，以作为多发性硬化症的潜在疗法。已证明来自多发性硬化症患者的公共卫生相关性T细胞对各种CNS抗原有反应，例如髓磷脂少突胶质细胞糖蛋白（MOG），髓磷脂碱性蛋白（MBP）和磷脂蛋白蛋白（PLP）；因此，对这些免疫反应的调节是MS治疗干预的目标。我们实验室的重点是开发一种新型的B细胞提供的基因治疗方法来诱导耐受性，尤其是在MS等自身免疫性疾病中。该平台已成功地用于MS的鼠模型中。本文中，我们将研究B细胞在动物模型中赋予耐受性的机制，然后我们将使用暴露于在体外用MBP-Ig构建体转导的B细胞的MS患者的T细胞将这些研究转化为诊所。这些研究将在临床上相关的模型中建立一种新型的与人T细胞的耐受性，并将继续将该项目转移到诊所，以此作为多发性硬化症的潜在疗法。