Lentiviral Vectors for the Treatment of Fanconi Anemia

描述（由申请人提供）：基于HIV-1的慢病毒载体正成为一种越来越有吸引力的将转基因整合到靶细胞中的方法。病媒正在进行或进入针对多种疾病的临床试验，包括遗传病、艾滋病和癌症。Rimedion和它的研究伙伴，印第安纳大学医学院提议开发一种新的慢病毒载体来治疗范可尼贫血。虽然载体结构将源自基于HIV-1的慢病毒系统，但载体将使用源自人类泡沫病毒的专有病毒包膜进行伪型。特异性目的1：通过人磷酸甘酸激酶启动子或延长因子1a启动子生成编码FANCA cDNA表达的重组慢病毒构建体，并测试转基因在小鼠FANCA -/-祖细胞和人FANCA缺陷淋巴母细胞样细胞系中的功能表达。具体目标2：使用GMP投诉方法优化慢病毒载体假型的生产，该慢病毒载体具有专有的PFV包膜。在这个第一阶段的应用中，临床载体的构建和优化的生产方法将最终确定。在II期，Rimedion将生成载体和完整的疗效和毒性数据，以纳入IND申请。这项工作将导致I期临床试验，使用新的载体转导来自范可尼贫血患者的自体外周血干细胞。正如在我们的应用中所讨论的那样，基因转移可以纠正这种疾病的血液学缺陷的原理证明已经很好地建立起来，使用先验载体系统进行基因转移的效率一直是开发商业产品的主要限制。Rimedion相信，所描述的新型载体系统将为这种遗传疾病提供一种治疗方法。

DESCRIPTION (provided by applicant): HIV-1 based lentiviral vectors are becoming an increasingly attractive means of integrating transgenes into target cells. Vectors are in or enterning clinical trials for a diverse group of ailments, including genetic disease, AIDS and cancer. Rimedion and its research partner, Indiana University School of Medicine proposes to develop a novel lentiviral vector for the treatment of Fanconi anemia. While the vector construct will be derived from HIV-1 based lentiviral system, the vector will be pseudotyped with a proprietary viral envelope derived from the human foamy virus. Specific Aim 1: Generate recombinant lentiviral constructs that encode the expression of the FANCA cDNA by the human phosphoglycerate kinase promoter or the elongation factor 1a promoter and test functional expression of the transgenes in murine Fanca -/- progenitors and human FANCA deficient lymphoblastoid cell lines. Specific Aim 2: Optimize production of a lentiviral vector pseudotyped with a proprietary PFV envelope using GMP complaint methodology. In this Phase I application, the clinical vector construct and optimized production methodology will finalized. In Phase II, Rimedion will generate vector and complete efficacy and toxicity data for inclusion in an IND filing. The work will lead to a Phase I clinical trial using the novel vector to transduce autologous peripheral blood stem cells from Fanconi anemia patients. As discussed in our application, the proof-of-principle has been well established that gene transfer can correct the hematologic defects of this disease, the efficiency of gene transfer using prior vector systems has been the major limitation to developing a commerical product. Rimedion believes the novel vector system described will provide a curative therapy for this genetic disease. PUBLIC HEALTH RELEVANCE: Gene therapy for Fanconi anemia has been successful in murine models but human cures have been limited by the low transduction efficiency of retroviral vectors. Rimedion will utilize a novel lentiviral vector combined with a proprietary foamy virus envelope to improve efficiency and decrease toxicity of gene transfer. Rimedion will optimize the vector and production methods in this application enabling a Phase I clinical trial.