Genome-Wide Computational Analysis Reveals Cardiomyocyte-Specific Transcriptional CIS-Regulatory Motifs that Enable Efficient Cardiac Gene Therapy.

Gene therapy is a promising emerging therapeutic modality for the treatment of cardiovascular diseases and hereditary diseases that afflict the heart. Hence, there is a need to develop robust cardiac-specific expression modules that allow for stable expression of the gene of interest in cardiomyocytes. We therefore explored a new approach based on a genome-wide bio-informatics strategy that revealed novel cardiac-specific cis-acting regulatory modules (CRMs). These transcriptional modules contained evolutionary conserved clusters of putative transcription factor binding sites that correspond to a 'molecular signature' associated with robust gene expression in the heart. We then validated these cardiac-specific CRMs in vivo using an adeno-associated viral vector serotype 9 (AAV9) that drives a reporter gene from a quintessential cardiac-specific ? myosin heavy chain (?MHC) promoter. Most de novo designed cardiac-specific CRMs resulted in a >10-fold increase in cardiac gene expression. The most robust CRMs enhanced cardiac-speciific transcription 70 to 100-fold. Expression was sustained and restricted to cardiomyocytes. We then combined the most potent CS-CRM4 with a synthetic heart and muscle-specific promoter (SPc5-12) and obtained a significant 20-fold increase in cardiac gene expression compared to the cytomegalovirus promoter. This study underscores the potential of rational vector design to improve the robustness of cardiac gene therapy.

Publication year:2015

Keywords:gene therapy, treatment, cardiovascular disease, hereditary disease, cardiomyocytes, CRMs

CSS-citation score:2

Accessibility:Open