Salivary Gland-Based Gene Therapy for Lysosomal Storage Diseases

描述(由申请人提供)：4R00DE018188-02的竞争性修订申请建议扩展父项目，以包括基于超声波破坏DNA相关微泡的非病毒基因传递方法。这种超声辅助基因转移(UAGT)的工作原理与母项目基于病毒的方法不同，但最终结果是，将基因转移到唾液腺是相同的。 因此，本附录被提议作为一个平行但同等的轨道，从PI的附属机构中引入一名具有近20年超声微泡技术经验的联合调查员。图1说明了父项目和提议的补充之间的关系。 通过协调执行母项目和拟议的补充研究计划，我们将能够利用在基于唾液腺的Fabry病基因治疗的生理和分子分析方面获得的现有专业知识。这将创造一种氛围，在严格控制的条件下，可以客观地比较病毒和非病毒基因转移方法的翻译潜力。随着基于唾液腺的基因治疗目前处于人类临床试验的第一阶段，如果要实现人类唾液腺的长期基因表达，这样的比较是必不可少的。 加强公共关系和L合作的主要机制是一名流动研究生的支持，该研究生将参与拟议补充项目的每一步。这一机制将把韦奕礼博士在构建和销毁超声波方面的专业知识联系起来 帕西诺博士基因治疗实验室的微泡造影剂。拟议的预算包括分配给这名在德雷克塞尔大学生物医学工程学院工作的研究生，让他经常延长访问匹兹堡进行动物工作和样本分析的时间。

DESCRIPTION (provided by applicant): This Competitive Revision Application for 4R00DE018188-02 proposes to expand the parent project to include a non-viral method of gene delivery based upon ultrasound destruction of DNA-associated microbubbles. This ultrasound-assisted gene transfer (UAGT) works differently from the viral-based methods of the parent project, but the end result, gene transfer to the salivary gland is the same. Therefore, this supplement is proposed as a parallel but equivalent track, bringing in a co-investigator from within the Pi's affiliated institution who has almost twenty years of experience with ultrasoundmicrobubble technology. Figure 1 illustrates the relationship of the parent project and proposed supplement. By executing the parent project and the proposed supplemental research plan in concert, we will be able to leverage the existing expertise acquired in the physiological and molecular analysis of salivary gland-based gene therapy for Fabry disease. This will create a climate where the translational potential of both viral and non-viral gene transfer methods can be objectively compared under tightly controlled conditions. With salivary gland-based gene therapy now in a Phase 1 human clinical trial, such comparisons are imperative if long-term gene expression in the salivary gland of humans is to be achieved. The major mechanism for cementing collaboration between the Pl and the Co-l is the support of a floating graduate student who will participate in every step of the proposed supplemental project. This mechanism will link Dr. Wheatley's expertise in the construction and destruction of ultrasound microbubble contrast agents with Dr. Passineau's gene therapy laboratory. The proposed budget includes allocation for this graduate student, based in the Drexel University School of Biomedical Engineering, to make frequent extended visits to Pittsburgh for animal work and sample analysis.