Utility of AAV serotypes for gene delivery in treatment of chronic joint disease

AAV 血清型在基因递送治疗慢性关节疾病中的应用

• 批准号: 8056625
• 负责人: Steven C Ghivizzani
• 金额: $ 30.9万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-09-15 至 2014-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8056625
• 关键词: Address; Affect; Animal Model; Animals; Arthritis; Biological; Biology; Capsid; Cellular Immunity; Chronic; Chronic Disease; Clinical; Complementary DNA; DNA biosynthesis; Data; Dependovirus; Development; Disease; Drug Kinetics; Effectiveness; Environment; Gene Delivery; Gene Expression; Gene Transduction Agent; Gene Transfer; Genes; Goals; Health; Human; Humoral Immunities; Immune; Immunity; Infection; Inflammation; Inflammatory; Intention; Intra-Articular Injections; Joints; Judgment; Link; Literature; Location; Mechanics; Mediating; Modeling; Molecular Profiling; Natural Immunity; Nude Rats; Pattern; Phenotype; Population; Recombinant adeno-associated virus (rAAV); Relative (related person); Reporting; Rheumatoid Arthritis; Safety; Serotyping; System; Systems Biology; Technology; Testing; Time; Tissues; Transgenes; Transgenic Organisms; Translations; Tropism; Tumor Necrosis Factor Receptor; Viral Vector; Woman; Work; adeno-associated viral vector; arthropathies; cellular transduction; clinical application; design; gene therapy; immunogenicity; phase 1 study; recombinant viral vector; research study; therapeutic transgene; transduction efficiency; transgene expression; vector

DESCRIPTION (provided by applicant): Arthritic diseases are chronic, crippling conditions for which there are no cures. We, and others, have shown that direct intra-articular injection of certain recombinant viral vectors can provide expression of therapeutic transgenes at levels sufficient to halt arthritis in animal models. Further, we have recently demonstrated that with the use of immunologically compatible vectors and cDNAs, exogenous transgenes can be expressed indefinitely in the joints of experimental animals. A primary barrier to clinical translation of gene therapies for chronic joint disease has been the lack of suitable vector systems. In this regard, recombinant adeno- associated virus (AAV) offers many advantages: it is nonpathogenic, of low immunogenicity, and enables persistent transgene expression in many applications. The capacity to cross-package, or pseudotype, the AAV2 vector has significantly expanded the versatility of this system, providing the opportunity to enhance transgenic expression as well as the potential to evade pre-existing immunity. Currently 9 AAV capsid serotypes are being developed as vectors for gene therapy applications. The recent development of double- stranded, self-complementary (sc) AAV vectors overcomes previous limitations associated with inefficient second strand DNA synthesis in articular tissues such that AAV vectors can be realistically considered for human application. Beyond the capacity of AAV vectors to deliver exogenous genes to joint tissue and achieve short-term functional expression, very little is known specifically of the biology of this system in the context of the articular environment. Thus, the goal of this project is to develop an understanding of the pharmacokinetics of AAV vector serotypes 1-9 intra-articularly to facilitate the clinical application of this technology in a safe, rational and effective manner. In Specific Aim 1, using a nude rat model to avoid issues of immune interference, we will establish local and systemic transgene delivery and expression profiles for AAV serotypes 1-9 following intra-articular administration. In Aim 2 we will perform similar experiments in an arthritic model to determine how morphologic changes associated with inflammatory disease affect AAV-mediated gene delivery and expression. For Aim 3, using immunologically competent animals we will determine the capacity to enhance or rescue AAV-mediated transgene expression by repeat vector administration and the use of alternate vector serotypes. In Aim 4 we will determine the impact of prior natural infection with wild type AAV2 on transgene delivery and expression of recombinant AAV vectors of the same and alternate serotypes. PUBLIC HEALTH RELEVANCE: Arthritic diseases are chronic, crippling conditions for which there are no cures. A primary barrier to clinical translation of gene therapies for chronic joint disease has been the lack of suitable vector systems. The goal of this project is to develop an understanding of the pharmacokinetics of the AAV vector serotypes intra-articularly to facilitate the clinical application of this technology in a safe, rational and effective manner.

描述（由申请人提供）：关节炎是慢性，残酷的疾病，没有治疗方法。我们和其他人已经表明，某些重组病毒载体的直接关节内注射可以提供足以阻止动物模型中关节炎的水平的治疗转基因表达。此外，我们最近证明，通过使用免疫学兼容的载体和cDNA，外源转基因可以在实验动物的关节中无限期地表达。慢性关节疾病基因疗法临床翻译的主要障碍是缺乏合适的载体系统。在这方面，重组腺相关病毒（AAV）具有许多优势：它是非致病性，免疫原性低，并且在许多应用中都可以持续的转基因表达。 AAV2矢量的交叉包装或假型的能力显着扩大了该系统的多功能性，从而提供了增强转基因表达的机会，并有可能逃避预先存在的免疫力。目前，正在开发9种AAV Capsid血清型作为基因治疗应用的载体。最近的双链，自相互（SC）AAV载体的发展克服了与关节组织中效率低下的第二链DNA合成相关的限制，因此可以在人类应用上实际考虑AAV量。除了AAV矢量向关节组织传递外源基因并实现短期功能表达的能力之外，在关节环境的背景下，该系统的生物学几乎没有知名度。因此，该项目的目的是建立对AAV矢量血清型的药代动力学1-9关节内的药代动力学，以促进该技术以安全，理性和有效的方式促进该技术的临床应用。在特定目标1中，使用裸大鼠模型避免免疫干扰问题，我们将在关节内给药后建立局部和系统的转基因传递和AAV血清型1-9的表达曲线。在AIM 2中，我们将在关节炎模型中进行类似的实验，以确定与炎症疾病相关的形态变化如何影响AAV介导的基因递送和表达。对于AIM 3，使用免疫学胜任的动物，我们将通过重复给药和使用替代矢量血清型来增强或拯救AAV介导的转基因表达的能力。在AIM 4中，我们将确定先前的自然感染对野生型AAV2对同一和替代血清型重组AAV载体的转基因递送和表达的影响。公共卫生相关性：关节炎是慢性，残酷的疾病，没有治疗方法。慢性关节疾病基因疗法临床翻译的主要障碍是缺乏合适的载体系统。该项目的目的是了解AAV矢量血清型的药代动力学的理解，以促进该技术以安全，理性和有效的方式促进该技术的临床应用。