Schröder Lena C, Hüttermann Leonard, Kliesow Remes Anca, Voran Jakob C, Hille Susanne, Sommer Wiebke, Lutter Georg, Warnecke Gregor, Frank Derk, Schade Dennis, Müller Oliver J
Department of Internal Medicine V, University of Kiel, Kiel, Germany.
German Centre for Cardiovascular Research, Partner Site Hamburg/Kiel/Lübeck, Kiel, Germany.
Gene Ther. 2025 Mar;32(2):154-162. doi: 10.1038/s41434-024-00511-8. Epub 2024 Dec 18.
Targeted gene delivery to vascular smooth muscle cells (VSMCs) could prevent or improve a variety of diseases affecting the vasculature and particularly the aorta. Thus, we aimed to develop a delivery vector that efficiently targets VSMCs. We selected engineered adeno-associated virus (AAV) capsids from a random AAV capsid library and tested the top enriched motifs in parallel screening through individual barcoding. This approach allowed us to distinguish capsids that only transduce cells based on genomic DNA (gDNA) from those also mediating transgene expression based on transcribed cDNA reads. After three rounds of selection on primary murine VSMCs (mVSMCs), we identified a novel targeting motif (RFTEKPA) that significantly improved transduction and gene expression efficiency over AAV9-wild type (WT) and increased expression in mVSMCs by 70% compared to the previously identified SLRSPPS peptide. Further analysis showed that the novel motif also improved expression in human aortic smooth muscle cells (HAoSMCs) and human aortic tissue ex vivo up to threefold compared to SLRSPPS and approximately 70-fold to AAV9-WT. This high cross-species transduction efficiency makes the novel capsid motif a potential candidate for future clinical application in vascular diseases.
将靶向基因递送至血管平滑肌细胞（VSMC）可预防或改善多种影响脉管系统尤其是主动脉的疾病。因此，我们旨在开发一种能有效靶向VSMC的递送载体。我们从随机的腺相关病毒（AAV）衣壳文库中筛选出工程化AAV衣壳，并通过单个条形码在平行筛选中测试高度富集的基序。这种方法使我们能够区分仅基于基因组DNA（gDNA）转导细胞的衣壳与那些还基于转录的cDNA读数介导转基因表达的衣壳。在对原代小鼠VSMC（mVSMC）进行三轮筛选后，我们鉴定出一种新型靶向基序（RFTEKPA），与AAV9野生型（WT）相比，其显著提高了转导和基因表达效率，与先前鉴定的SLRSPPS肽相比，mVSMC中的表达增加了70%。进一步分析表明，与SLRSPPS相比，该新型基序在人主动脉平滑肌细胞（HAoSMC）和离体人主动脉组织中的表达也提高了三倍，与AAV9-WT相比提高了约70倍。这种高跨物种转导效率使新型衣壳基序成为未来血管疾病临床应用的潜在候选者。
