Authors: Marta Adamiak, Divya Jha, Yaxuan Liang, Prabhu Mathiyalagan, Neha Agarwal, Erik Kohlbrenner, Elena Chepurko, Dongtak Jeong, Delaine Ceholski, Nicole Dubois, Roger Hajjar, Susmita Sahoo
Published: 2019-10-16
DOI: 10.1161/res.125.suppl_1.170
Source: Full article
Adeno-associated viruses (AAVs) are promising therapeutic tools for gene delivery to the heart. However, pre-existing antibodies (NAbs) to many cardiotropic AAV serotypes naturally present in humans pose a critical challenge for the translation of gene therapies to clinical applications. Here, we describe the use of exosomal AAVs (eAAV) as a robust heart gene delivery system that improves transduction efficiency while protecting from pre-existing immunity to the viral capsid. To obtain eAAV specimens from conditioned medium from AAV-producing HEK-293T cells, we have developed a state-of-the-art multi-step ultracentrifugation strategy. We demonstrated through electron microscopy-based visualization, size distribution measurements and distribution of AAV genomes in post-centrifugation iodixanol gradients, that our purification process enables isolation of eAAVs with high purity and minimal contamination with standard AAVs. Efficiency of heart targeting was then evaluated for eAAV9 or eAAV6 and standard AAV9 or AAV6 in human cardiomyocytes (hCMs)