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（由应用程序提供）：1型糖尿病（T1D）的特征是自身免疫介导的胰岛素产生的破坏了兰格汉斯胰岛的细胞。我们的工作和其他在鼠模型中的工作表明，将免疫调节分子的基因转移到内源性胰腺中的体内或体外胰岛移植物中的细胞，分别有效抑制自身免疫并促进胰岛移植耐受性。我们一直在使用腺相关病毒（AAV）载体基因转移来操纵体内和体外细胞的免疫原性。 AAV媒介技术已经快速先进，临床试验表明该基因输送策略的成功应用。考虑到这一点，我们的R21提案的目标是开发AAV向量，并在体内和体外对人类细胞的向量增加。将采取两种约束的方法。首先，我们将采用一组野生型AAV CAPSID蛋白来识别那些在体外和体内模型中有效地翻译人类细胞的衣壳。其次，将对工程师的新型AAV帽质变体做出努力，该变体在体内和体外有选择地翻译人的细胞，并逃避人AAV中和抗体。在这里，与定向进化的AAV Capsid基因的“ DNA洗牌”将用于开发新颖的细胞特异性AAV衣壳。实验还将包括测试AAV载体处理的有效性，以阻止体内人类细胞的免疫介导的破坏。总之，这项工作将更好地理解对人类细胞的AAV衣壳的翻译特性，并将识别/生成可以在诊所直接测试的衣壳。这样，将采取初始步骤，即我们使用AAV矢量来建立人类T1D的细胞特异性和/或胰岛移植耐受性。