Basset Justine, Seraffin Alexandra, Ratelade Julien, Dickx Yohann, Benedyk Tomasz, Sarek Grzegorz, Jégu Teddy, Epstein Alberto L
EG427, 29 Rue du Faubourg Saint Jacques, 75014 Paris, France.
Int J Mol Sci. 2025 May 21;26(10):4941. doi: 10.3390/ijms26104941.
Non-replicative herpes simplex virus type 1 (nrHSV-1) vectors are promising delivery vehicles for gene therapy due to their large DNA payload capacity and ability to infect a broad range of cell types. However, the genomic deletions made to generate such nrHSV-1 vectors can result in undersized genomes that trigger genomic instability-including rearrangements and size extensions-compromising their therapeutic potential. This study investigates the stabilization of undersized nrHSV-1 vectors through the insertion of stuffer DNA segments. We assess genomic stability, productivity, toxicity, and transgene expression in vitro and in vivo. Our findings demonstrate that nrHSV-1 can accommodate variations in genome size up to 5-6% and highlight the importance of maintaining a genome size close to that of the wild-type HSV-1 for enhanced genomic stability and sustained transgene expression without adverse effects. This strategy offers a promising approach for optimizing nrHSV-1 vectors for clinical applications.
非复制型单纯疱疹病毒1型（nrHSV-1）载体因其较大的DNA承载能力和感染多种细胞类型的能力，是基因治疗中很有前景的递送载体。然而，为产生此类nrHSV-1载体而进行的基因组缺失可能导致基因组过小，从而引发基因组不稳定，包括重排和大小扩展，损害其治疗潜力。本研究通过插入填充DNA片段来研究过小的nrHSV-1载体的稳定性。我们在体外和体内评估了基因组稳定性、生产力、毒性和转基因表达。我们的研究结果表明，nrHSV-1能够容纳高达5-6%的基因组大小变化，并强调了保持基因组大小接近野生型HSV-1对于增强基因组稳定性和持续转基因表达且无不良影响的重要性。该策略为优化nrHSV-1载体用于临床应用提供了一种很有前景的方法。
