In Vivo Editing for Hemophilia Gene Therapy

项目摘要 最近的研究表明，血友病的基因治疗具有特殊的前景，其结果是 通过腺相关载体(AAV)的肝脏转导，长期纠正因子IX缺乏症。在……里面 尽管这是基因治疗领域历史上的分水岭事件，但目前仍存在最充分的障碍。 实施血友病基因治疗。AAV对较大基因结构的包装能力有限 实际上混淆了该载体用于第VIII因子校正和与载体相关的毒性 值得注意的是体内转导肝脏的背景。我们建议开发一种新的矢量法 这解决了这些限制。值得注意的是，我们最近重定向腺病毒载体(Ad)的策略已经允许 减轻载体相关的肝毒性和体内转导肺内皮细胞 证明该位点可以作为一个完全有效的平台来重建缺乏的血清蛋白。 此外，具有多个删除的新一代广告可以进行修改，以纳入更多范围的 与血友病全谱基因治疗相关的基因结构和更大的有效载荷。我们 建议利用与广告技术完全相称的CRISPR/CAS9系统， 利用我们的肺内皮靶向Ad，实现纠正血友病基因的稳定整合 在肺内皮细胞内。因此，我们的新策略提供了稳定的基因校正的可能性 以避免因子IX和因子VIII血友病的潜在载体相关毒性的方式。 成功完成本可行性证明阶段1中的目标将为以下工作奠定基础 血友病矫正性基因治疗在最广泛的患者中的应用并提供理论基础 对于我们在第二阶段的销售线索优化和选择方面非常新颖的方法的翻译开发。我们的 在新基因疗法的成功从工作台到病床的翻译方面，团体拥有无与伦比的记录 战略。我们实现缺陷血友病因子稳定表达的策略 肺血管来源显然代表了一种高度原创性的方法。除了绕过这个问题 在载体介导的肝毒性中，腺病毒载体的扩展包装能力也是可行的 因子VIII以及因子IX缺乏症的基因治疗。为了加快我们的方法的转换，我们 将利用来自非人灵长类腺病毒的腺病毒载体，并将专门使用大猩猩Ad 载体(Gad)通过绕过预先形成的腺病毒免疫和 在治疗应用中表现出优异的性能特点。功效研究概况 这里将构成这一新方法翻译开发的理论基础，以及对 建立肺内皮细胞作为来源来提供分泌因子是一种重要的策略 基于缺乏血清因素和其他潜在因素的遗传性遗传性疾病的范围 治疗应用。

Project Summary The exceptional promise of gene therapy for hemophilia has been shown in recent studies that result in long term correction of factor IX deficiency, via hepatic transduction with an adeno-associated vector (AAV). In spite of this watershed event in the history of the gene therapy field, there presently exist barriers to the fullest implementation of hemophilia gene therapy. The limited packaging capacity of AAV for larger gene constructs practically confounds the use of this vector for factor VIII correction and vector-related toxicities have been noteworthy for the context of in vivo transduction of the liver. We propose to develop a novel vector approach that addresses these limitations. Of note, our recent strategies to retarget adenoviral vectors (Ad) have allowed mitigation of vector-associated liver toxicities and in vivo transduction of the pulmonary endothelium that has demonstrated that this site can serve as a fully effective platform to reconstitute deficient serum proteins. Moreover, new generation Ads with multiple deletions can be modified to incorporate an expanded range of gene constructs and larger payloads relevant to gene therapy for the full spectrum of hemophilia disorders. We propose to utilize the CRISPR/Cas 9 system which is fully commensurate with Ad technology, in conjunction with our pulmonary endothelial-targeted Ad, to achieve stable incorporation of corrective hemophilia genes within the pulmonary endothelium. Our novel strategy thereby offers the potential for stable genetic correction in a manner that circumvents potential vector-associated toxicities for both factor IX and factor VIII hemophilia. Successful completion of the aims within this proof of feasibility Phase 1 STTR will set the stage for the application of corrective gene therapy for hemophilia to the widest context of patients and provide the rationale for translational development of our highly novel approach into lead optimization and selection in Phase II. Our groups have unparalleled track records for successful bench-to-bed translation of novel gene therapy strategies. Our strategy to accomplish stable expression of deficient hemophilia factors deriving from a pulmonary vascular source clearly represents a highly original approach. In addition to circumventing the issue of vector-mediated hepatotoxicities, the expanded packaging capacity of adenovirus vectors also feasibilizes gene therapy for factor VIII, as well as factor IX deficiency disorders. To hasten translation of our approach, we will utilize adenoviral vectors derived from non-human primate adenoviruses and will specifically use gorilla Ad vectors (GAd) that address a pivotal human use barrier by circumventing preformed adenoviral immunity and are show excellent performance characteristics in therapeutic applications. The profile of efficacy studies herein will constitute the rationale for translational development of this novel approach as well as a leap to establishing the pulmonary endothelium as a source to provide secreted factors as an important strategy for the range of inherited genetic disorders based upon deficient serum factors as well as other potential therapeutic applications.