Liu Y B, Xu B C, Chen Y T, Yuan X, Liu J Y, Liu T, Du G Z, Jiang W, Yang Y, Zhu Y, Chen L J, Ding B S, Wei Y Q, Yang L
Department of Cardiology and Laboratory of Gene Therapy for Heart Diseases, State Key Laboratory of Biotherapy, West China Hospital, Sichuan University and Collaborative Innovation Center for Biotherapy, Chengdu, Sichuan, China.
Laboratory of Aging Research and Cancer Drug Target, State Key Laboratory of Biotherapy and Cancer Center, National Clinical Research Center for Geriatrics, West China Hospital, Sichuan University, Chengdu, Sichuan, China.
Mol Ther Methods Clin Dev. 2021 Jun 4;22:148-161. doi: 10.1016/j.omtm.2021.05.015. eCollection 2021 Sep 10.
Cardiac endothelial cells (ECs) are important targets for cardiovascular gene therapy. However, the approach of stably transducing ECs using different vectors, including adeno-associated virus (AAV), remains unexamined. Regarding this unmet need, two AAV libraries from DNA shuffling and random peptide display were simultaneously screened in a transgenic mouse model. Cardiac ECs were isolated by cell sorting for salvage of EC-targeting AAV. Two AAV variants, i.e., EC71 and EC73, enriched in cardiac EC, were further characterized for their tissue tropism. Both of them demonstrated remarkably enhanced transduction of cardiac ECs and reduced infection of liver ECs in comparison to natural AAVs after intravenous injection. Significantly, persistent transgene expression was maintained in mouse cardiac ECs for at least 4 months. The EC71 vector was selected for delivery of the endothelial nitric oxide synthase (eNOS) gene into cardiac ECs in a mouse model of myocardial infarction. Enhanced eNOS activity was observed in the mouse heart and lung, which was correlated with partially improved cardiac function. Taken together, two AAV capsids were evolved with more efficient transduction in cardiovascular endothelium , but their endothelial tropism might need to be further optimized for practical application to cardiac gene therapy.
心脏内皮细胞（ECs）是心血管基因治疗的重要靶点。然而，使用包括腺相关病毒（AAV）在内的不同载体稳定转导ECs的方法仍未得到研究。针对这一未满足的需求，在转基因小鼠模型中同时筛选了来自DNA改组和随机肽展示的两个AAV文库。通过细胞分选分离心脏ECs，以挽救靶向EC的AAV。进一步对在心脏EC中富集的两个AAV变体EC71和EC73的组织嗜性进行了表征。与静脉注射后的天然AAV相比，它们在心脏ECs中的转导均显著增强，而在肝脏ECs中的感染减少。值得注意的是，小鼠心脏ECs中持续的转基因表达维持了至少4个月。在心肌梗死小鼠模型中，选择EC71载体将内皮型一氧化氮合酶（eNOS）基因递送至心脏ECs。在小鼠心脏和肺中观察到eNOS活性增强，这与心脏功能的部分改善相关。综上所述，两个AAV衣壳在心血管内皮细胞中进化出了更有效的转导能力，但它们的内皮嗜性可能需要进一步优化，以便实际应用于心脏基因治疗。
