Lentiviral Vectors for the Treatment of Fanconi Anemia

描述（由申请人提供）：基于HIV-1的慢病毒载体正在成为将转基因整合到靶细胞中的越来越有吸引力的手段。载体正在进行或即将进入各种疾病的临床试验，包括遗传病、艾滋病和癌症。Rimedion和它的研究伙伴，印第安纳州大学医学院提出开发一种新的慢病毒载体用于治疗范可尼贫血。虽然载体构建体将衍生自基于HIV-1的慢病毒系统，但载体将用衍生自人泡沫病毒的专有病毒包膜进行假型化。具体目标1：生成重组慢病毒构建体，其编码通过人磷酸甘油酸激酶启动子或延伸因子1a启动子表达的FANCA cDNA，并检测转基因在鼠Fanca -/-祖细胞和人FANCA缺陷型淋巴母细胞系中的功能表达。具体目标2：使用符合GMP要求的方法优化用专有PFV包膜假型化的慢病毒载体的生产。在此I期申请中，临床载体构建和优化的生产方法将最终确定。在第二阶段，Rimedion将生成载体和完整的疗效和毒性数据，以纳入IND文件。这项工作将导致一项I期临床试验，使用新的载体从范可尼贫血患者中培养自体外周血干细胞。如在我们的申请中所讨论的，已经充分确立了基因转移可以纠正这种疾病的血液学缺陷的原理证明，使用现有载体系统的基因转移效率一直是开发药物产品的主要限制。Rimedion认为，所描述的新型载体系统将为这种遗传疾病提供治愈性疗法。 公共卫生相关性：范可尼贫血的基因治疗已经在小鼠模型中取得成功，但人类的治疗受到逆转录病毒载体转导效率低的限制。Rimedion将利用一种新型的慢病毒载体与专有的泡沫病毒包膜相结合，以提高效率并降低基因转移的毒性。Rimedion将优化该应用中的载体和生产方法，从而实现I期临床试验。

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.