Repurposing an endogenous human mRNA transfer system as a gene therapy tool

这个项目的目的是绘制出一个自然存在的系统的可行性和性能，当重新用于传递重组RNA有效载荷时，mRNA在人体内的转移。该系统涉及一种蛋白质，该蛋白质在翻译后与其编码的mRNA结合，然后形成一个颗粒，将该mRNA隔离，之后该颗粒被包裹在囊泡内，在细胞释放和病毒样转移到靶细胞之前形成多泡体（MVB），在靶细胞中释放有效载荷mRNA进行后续翻译。基因治疗方法最近面临挑战，部分原因是由于用于基因转移的病毒载体的免疫原性产生的不利影响。Kumin等人（2021）报告称，在2021年进行的149项AAV基因治疗临床试验中，平均有35%与治疗后出现的严重不良事件相关。在这个项目中，将对Protein X mRNA进行修饰，以测试其整合重组RNA元素的能力。不同大小和序列的插入元件将通过工程蛋白X系统测试其从包装细胞转移到靶细胞的能力。预计蛋白- x系统的免疫原性将明显低于基于病毒的系统。

This project aims to map out the feasibility and performance of a naturally-ocurring system for mRNA transfer in humans when repurposed to deliver recombinant RNA payloads. The system involves a single protein that, upon translation, associates with its encoding mRNA then forms a particle that sequesters that mRNA, after which the particle is enveloped within vesicles that bud into a multivesicular body (MVB) before cellular release and virus-like transfer to target cells in which the payload mRNA is released for subsequent translation.Gene therapy approaches have recently faced challenges due in part to adverse effects arising from the immunogenicity of the viral vectors used for tgene transfer. Kumin et al. (2021) report that, on average, 35% of 149 2021 AAV gene therapy clinical trials were associated with treatment-emergent serious adverse events.In this project the Protein X mRNA will be modified to test its capacity to incorporate recombinant RNA elements. Inserted elements of differing size and sequence will be tested for their ability to be transferred from packaging cells to target cells via the engineerred Protein X system. It is anticipated that the immunogenicity of a Protein-X system will be significantly lower than that of virus-based systems.