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Development of Human beta cell-specific AAV Vectors for Type I Diabetes

开发用于 I 型糖尿病的人类 β 细胞特异性 AAV 载体

基本信息

批准号：
8663188
负责人：
RICHARD J SAMULSKI
金额：
$ 19万
依托单位：
UNIV OF NORTH CAROLINA CHAPEL HILL
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-06-01 至 2015-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8663188
关键词：
Autoimmune Process Autoimmune Responses Autoimmunity Beta Cell CD4 Positive T Lymphocytes CD8B1 gene Capsid Capsid Proteins Cells Clinic Clinical Research Clinical Trials DNA Shuffling Dependovirus Dose Engineering Exhibits Foundations Gene Delivery Gene Targeting Gene Transfer Genes Goals Graft Tolerance Hereditary Disease Human Human Development Immune Immune Tolerance Immunotherapy In Vitro Inbred NOD Mice Insulin Insulin-Dependent Diabetes Mellitus Interleukin-2 Islet Cell Islets of Langerhans Islets of Langerhans Transplantation Mediating Mind Modeling Modification Mus Nature Pancreas Pathology Patients Property Regulatory T-Lymphocyte Self Tolerance Serotyping Serum T-Lymphocyte Technology Testing Therapeutic Effect Tissues Transplantation Tolerance Tropism Variant Work adeno-associated viral vector base cell type cellular transduction clinical application cytokine directed evolution efficacy testing immunogenicity immunoregulation in vivo in vivo Model interest islet neutralizing antibody novel promoter public health relevance research study tissue tropism transduction efficiency transgene expression vector

项目摘要

DESCRIPTION (provided by applicant): Type 1 diabetes (T1D) is characterized by the autoimmune-mediated destruction of the insulin producing ¿ cells of the islets of Langerhans. Our work and that of others in murine models has shown that transferring genes for immunoregulatory molecules to ¿ cells in vivo in the endogenous pancreas or in vitro in islet grafts, effectively suppresses autoimmunity and promotes islet transplantation tolerance, respectively. We have been using adeno-associated virus (AAV) vector gene transfer to manipulate the immunogenicity of murine ¿ cells in vivo and in vitro. AAV vector technology has rapidly advanced and clinical trials have demonstrated successful application of this gene delivery strategy. With this in mind, the goal of our R21 proposal is to develop AAV vectors with increased tropism for human ¿ cells in vivo and in vitro. A two-pronged approach will be taken. First, we will employ a panel of wild-type AAV capsid proteins to identify those capsids that efficiently transduce human ¿ cells in vitro, and in an in vivo model. Secondly, an effort will be made to engineer novel AAV capsid variants that selectively transduce human ¿ cells in vivo and in vitro, and which evade human AAV neutralizing antibodies. Here "DNA shuffling" of AAV capsid genes combined with directed evolution will be used to develop novel ¿ cell-specific AAV capsids. Experiments will also include testing the efficacy of AAV vector treatment to block immune-mediated destruction of human ¿ cells in vivo. Together, this work will provide a better understanding of the transduction properties of AAV capsids for human ¿ cells, and will identify/generate capsids that can be directly tested in the clinic. In this way, initial steps wil be taken towards our long-term goal of using AAV vectors to establish ¿ cell-specific and/or islet graft tolerance for human T1D.

描述（由申请人提供）：1型糖尿病（T1D）的特征是自身免疫介导的胰岛胰岛细胞的胰岛素产生破坏。我们和其他人在小鼠模型中的工作表明，将免疫调节分子的基因转移到体内内源性胰腺或体外胰岛移植物的细胞中，分别有效抑制自身免疫并促进胰岛移植耐受。我们一直在使用腺相关病毒（AAV）载体基因转移来操纵小鼠细胞体内和体外的免疫原性。 AAV 载体技术迅速发展，临床试验已证明这种基因递送策略的成功应用。考虑到这一点，我们 R21 提案的目标是开发在体内和体外对人类细胞具有增强趋向性的 AAV 载体。将采取双管齐下的方法。首先，我们将使用一组野生型 AAV 衣壳蛋白来鉴定那些能够在体外和体内模型中有效转导人类细胞的衣壳。其次，将努力设计新型 AAV 衣壳变体，在体内和体外选择性转导人类细胞，并逃避人类 AAV 中和抗体。这里，AAV 衣壳基因的“DNA 改组”与定向进化相结合，将用于开发新型细胞特异性 AAV 衣壳。实验还将包括测试 AAV 载体治疗在体内阻止免疫介导的人体细胞破坏的功效。总之，这项工作将有助于更好地了解 AAV 衣壳对人类细胞的转导特性，并将识别/生成可在临床中直接测试的衣壳。通过这种方式，我们将朝着使用 AAV 载体建立人类 T1D 的细胞特异性和/或胰岛移植耐受性的长期目标迈出第一步。