Facilitate by-stander effects by EV-mRNA cargo in AAV gene therapy for MPS IIIC

项目摘要 MPS IIIC是一种毁灭性的溶酶体储存性疾病，目前还没有明确的治疗方法。致信地址 这一未得到满足的需求，这项拟议的项目是开发一种新的有效的基因治疗产品来治疗MPS IIIC 使用AAV载体来定位根本原因。值得注意的是，MPS IIIC的神经病理是全球性的，涉及 HGSNAT是一种跨溶酶体膜蛋白，与大多数 溶酶体酶，不能被邻近细胞分泌和摄取。因此，最佳的治疗方法 好处是需要靶向尽可能多的细胞，这可以通过高剂量的载体传递实现，例如 相比之下，最近批准Zolgensma用于SMA.25的研究表明，基因治疗对 涉及一种可分泌蛋白质的MPS疾病需要显著较低的载体剂量。高媒介剂量 由于载体生产的放大能力，AAV基因治疗的翻译一直是一个重大挑战 供人类使用。众所周知，所有细胞都能持续释放细胞外小泡(EV)并进行通讯 通过电动汽车交通交换大分子。EV细胞间相互作用的特性提供了一种 潜在的工具，以缓解这一挑战的rAAV基因治疗的翻译。我们假设公司成立于 EV-mRNA包装在基因治疗载体中的信号将对正常非分泌产生旁观者效应 蛋白通过含有mRNAs的EVS。为了验证这一假设，我们将构建rAAV载体，以表达 HHGSNAT和含有不同的EV包装信号，并识别EV-mRNA的有效信号 正在装车。目的是建立一种有效的rAAV-hHGSNAT基因治疗MPS IIIC的方法。数据 也可能为开发基因疗法来治疗其他涉及非 隐秘的转基因产品。我们相信该项目非常适合R21高风险/高影响资金 机制。

Project Summary MPS IIIC is a devastating lysosomal storage disease (LSD), for which there is no definitive treatment. To address this unmet need, this proposed project is to develop a novel effective gene therapy product for treating MPS IIIC using AAV vector to target the root cause. Notably, the neuropathologies in MPS IIIC are global, involving the entire nervous system, and HGSNAT is a trans-lysosomal-membrane protein, which, unlike the majority of lysosomal enzymes, cannot be secreted and taken-up by neighboring cells. Therefore, optimal therapeutic benefits requires targeting as many cells as possible, which can be achieved by high-dose vector delivery, such as recently approved Zolgensma for SMA.25 In comparison, studies have demonstrated that gene therapy for MPS disorders involving a secretable protein required significantly lower vector doses. The high vector doses has been a major challenge in translation of AAV gene therapy due to the scale-up capacity of vector production for human application. All cells are known to continuously release extracellular vesicles (EVs) and communicate by exchanging large molecules via EV traffic. The demonstrated properties of EV intercellular interaction offer a potential tool to ease this challenge to translation of rAAV gene therapy. We hypothesize that incorporation of EV-mRNA packaging signals in gene therapy vectors will engender by-stander effects for normally non-secreted protein via the mRNA-containing EVs. To test this hypothesis, we will construct rAAV vectors that express hHGSNAT and contain different putative EV packaging signals, and identify effective signals for EV-mRNA loading. The goal is to develop an effective rAAV-hHGSNAT gene delivery approach for treating MPS IIIC. Data generated may also provide powerful tools for developing gene therapy to treat other diseases involving non- secreted transgene products. We believe this project is ideally suited to the R21 high risk/high impact funding mechanism.