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

在 MPS IIIC 的 AAV 基因治疗中通过 EV-mRNA 货物促进旁观者效应

基本信息

批准号：
10040153
负责人：
HAIYAN FU
金额：
$ 42.76万
依托单位：
UNIV OF NORTH CAROLINA CHAPEL HILL
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-09-01 至 2023-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10040153
关键词：
Address Binding Proteins Cell Culture Techniques Cells Code Data Development Disease Dose Enzymes Family Fibroblasts Funding Mechanisms GAG Gene Gene Delivery Gene Transduction Agent Goals Human In Situ In Vitro Lysosomal Storage Diseases Mediating Membrane Proteins Messenger RNA Nervous system structure Patients Plant Roots Plasmid Cloning Vector Plasmids Production Property Proteins Recombinant adeno-associated virus (rAAV)Ribosomes Signal Transduction Signaling Protein Skin Testing Therapeutic Transgenes Translations Vectorial capacity Viral Vector adeno-associated viral vector cellular transduction clinical application experience extracellular vesicles gene therapy high risk human disease in vitro testing intercellular communication neurogenetics neuropathology novel novel strategies protein expression scale up success tool vector

项目摘要

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.

项目概要 MPS IIIC 是一种毁灭性的溶酶体贮积病 (LSD)，目前尚无明确的治疗方法。致地址针对这一未满足的需求，本提议的项目是开发一种新型有效的基因治疗产品来治疗 MPS IIIC 使用 AAV 向量来定位根本原因。值得注意的是，MPS IIIC 的神经病理学是全球性的，涉及整个神经系统，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吸收。因此，最佳治疗好处需要靶向尽可能多的细胞，这可以通过高剂量载体递送来实现，例如最近批准的 Zolgensma 用于治疗 SMA。25 相比之下，研究表明基因治疗涉及可分泌蛋白的 MPS 疾病需要显着降低的载体剂量。高载体剂量由于载体生产能力的扩大，AAV 基因治疗的转化一直是一个重大挑战供人类应用。众所周知，所有细胞都会持续释放细胞外囊泡 (EV) 并进行交流通过电动汽车交通交换大分子。 EV 细胞间相互作用的已证实特性提供了缓解 rAAV 基因治疗翻译挑战的潜在工具。我们假设合并基因治疗载体中的 EV-mRNA 包装信号将对正常非分泌的细胞产生旁观者效应通过含有 mRNA 的 EV 产生蛋白质。为了检验这个假设，我们将构建表达的 rAAV 载体 hHGSNAT 包含不同的推定 EV 包装信号，并识别 EV-mRNA 的有效信号加载中。目标是开发一种有效的 rAAV-hHGSNAT 基因递送方法来治疗 MPS IIIC。数据产生的结果还可能为开发基因疗法提供强大的工具，以治疗涉及非分泌的转基因产物。我们相信该项目非常适合 R21 高风险/高影响力资金机制。