Wang Jianan, Jonker Timo, Cervera-Barea Aina, Dong Zhenyu, Visser Ruud N, Birza Evelien E, Boender Arie R, Klerk Mischa, Yang Yuting, Visser Joyce, Jansen Marlijn S, Grootswagers Tom C, Bart Cindy I, de Jager Saskia C A, Schrödel Silke, Thirion Christian, Kirzner Osne F, Tan Hanno L, de Vries Antoine A F, Sluijter Joost P G, Neef Klaus, de Groot Joris R, Christoffels Vincent M, Boink Gerard J J
Department of Medical Biology, Amsterdam Cardiovascular Sciences, Amsterdam University Medical Centers, University of Amsterdam, 1105 AZ Amsterdam, the Netherlands.
Laboratory for Experimental Cardiology, Regenerative Medicine Center Utrecht, Circulatory Health Research Center, University Medical Center Utrecht, Utrecht University, 3584 CX Utrecht, the Netherlands.
Mol Ther Methods Clin Dev. 2025 Jul 15;33(3):101532. doi: 10.1016/j.omtm.2025.101532. eCollection 2025 Sep 11.
Cardiac gene therapy using adeno-associated viral (AAV) vectors holds great promise for treating heart diseases but would benefit from more potent AAV vectors. Vectors based on the AAV serotypes 6 and 9 have been used in pre-clinical gene therapy studies, yet the therapeutic outcomes varied depending on the experimental model and delivery route used. Here, we evaluated the transduction efficiency of AAV6, AAV9, and AAV9-derived MyoAAVs for local cardiac delivery. Vectors were tested in neonatal rat ventricular myocytes, and subsequently in mouse hearts by direct intramyocardial injection. Vector genome levels, mRNA expression levels, and fluorescence were measured. The AAV6 and AAV9 vectors were further validated in porcine hearts, human-induced pluripotent stem-cell-derived cardiomyocytes, and human atrial myocardial slices. In both rat cardiomyocytes and mouse hearts, AAV6 exhibited the highest transduction efficiency. Direct comparison of the AAV6 and AAV9 vectors in porcine and human models confirmed that AAV6 is more potent. In conclusion, AAV6 vectors are superior to AAV9 and its derivative vectors for cardiac transduction by direct intramyocardial injection. In addition, the transduction efficiency correlates with and assays, thereby facilitating the development of more potent AAV variants for cardio-selective delivery methods.
使用腺相关病毒（AAV）载体进行心脏基因治疗在治疗心脏病方面具有巨大潜力，但需要更有效的AAV载体才能取得更好效果。基于AAV血清型6和9的载体已用于临床前基因治疗研究，然而治疗结果因所使用的实验模型和递送途径而异。在此，我们评估了AAV6、AAV9和AAV9衍生的肌特异性AAV（MyoAAV）用于局部心脏递送的转导效率。载体在新生大鼠心室肌细胞中进行测试，随后通过直接心肌内注射在小鼠心脏中进行测试。测量了载体基因组水平、mRNA表达水平和荧光。AAV6和AAV9载体在猪心脏、人诱导多能干细胞衍生的心肌细胞和人心房心肌切片中进一步得到验证。在大鼠心肌细胞和小鼠心脏中，AAV6均表现出最高的转导效率。在猪和人类模型中对AAV6和AAV9载体进行直接比较证实，AAV6更有效。总之，通过直接心肌内注射进行心脏转导时，AAV6载体优于AAV9及其衍生载体。此外，转导效率与[此处原文缺失相关检测内容]检测相关，从而有助于开发更有效的用于心脏选择性递送方法的AAV变体。
