AAV-mediated gene therapy has become a promising therapeutic strategy for chronic diseases. Its clinical utilization, however, is limited by the potential risk of off-target effects. In this work we attempt to overcome this challenge, hypothesizing that cardiac ion channel-specific ligands could be fused onto the AAV capsid, and narrow its tropism to cardiac myocytes. We successfully fused the cardiac sodium channel (Na_v1.5)-binding toxin Anthopleurin-B onto the AAV2 capsid without compromising virus integrity, and demonstrated increased specificity of cardiomyocyte attachment. Although virus attachment to Na_v1.5 did not supersede the natural heparan-mediated virus binding, heparan-binding ablated vectors carrying Anthopleurin-B eliminated hepatic and other extracardiac gene transfer, while preserving cardiac myocyte gene transfer. Virus binding to the cardiac sodium channel transiently decreased sodium current density, but did not cause any arrhythmias. Our findings expand the knowledge of attachment, infectivity, and intracellular processing of AAV vectors, and present an alternative strategy for vector retargeting.

AAV介导的基因治疗已成为一种有前途的慢性疾病治疗策略。但是，其临床利用受到脱靶效应的潜在风险限制。在这项工作中，我们尝试克服这一挑战，并假设可以将心脏离子通道特异性配体融合到AAV衣壳上，并缩小其向心肌细胞的向性。我们成功地将结合心脏钠通道（Na _v 1.5）的毒素Anthopleurin-B融合到AAV2衣壳上，而没有损害病毒的完整性，并证明了增加的心肌细胞附着特异性。尽管病毒依附于Na _v1.5没有取代天然乙酰肝素介导的病毒结合，携带蒽醌-B的乙酰肝素结合消融载体消除了肝和其他心外基因的转移，同时保留了心肌细胞基因的转移。病毒与心脏钠通道的结合会暂时降低钠电流密度，但不会引起任何心律不齐。我们的发现扩展了AAV载体的附着，感染性和细胞内加工的知识，并提出了载体重新靶向的另一种策略。