title: Genome-wide computational analysis reveals cardiomyocyte-specific transcriptional Cis-regulatory motifs that enable efficient cardiac gene therapy.
abstract: 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 bioinformatics strategy that revealed novel cardiac-specific cis-acting regulatory modules (CS-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 CS-CRMs in vivo using an adeno-associated viral vector serotype 9 that drives a reporter gene from a quintessential cardiac-specific α-myosin heavy chain promoter. Most de novo designed CS-CRMs resulted in a >10-fold increase in cardiac gene expression. The most robust CRMs enhanced cardiac-specific 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.
authors.affiliation: 1] Department of Gene Therapy & Regenerative Medicine, Free University of Brussels (VUB), Brussels, Belgium [2] Center for Molecular & Vascular Biology, Department of Cardiovascular Sciences, University of Leuven, Leuven, Belgium.
authors.forename: Melvin Y
authors.initials: MY
authors.lastname: Rincon
authors.affiliation: Department of Gene Therapy & Regenerative Medicine, Free University of Brussels (VUB), Brussels, Belgium.
authors.forename: Shilpita
authors.initials: S
authors.lastname: Sarcar
authors.affiliation: Center for Molecular & Vascular Biology, Department of Cardiovascular Sciences, University of Leuven, Leuven, Belgium.
authors.forename: Dina
authors.initials: D
authors.lastname: Danso-Abeam
authors.affiliation: Nuclear Medicine Department, UZ Brussel & In vivo Cellular and Molecular Imaging Lab, Free University of Brussels (VUB), Brussels, Belgium.
authors.forename: Marleen
authors.initials: M
authors.lastname: Keyaerts
authors.affiliation: 1] Department of Gene Therapy & Regenerative Medicine, Free University of Brussels (VUB), Brussels, Belgium [2] Center for Molecular & Vascular Biology, Department of Cardiovascular Sciences, University of Leuven, Leuven, Belgium.
authors.forename: Janka
authors.initials: J
authors.lastname: Matrai
authors.affiliation: 1] Department of Gene Therapy & Regenerative Medicine, Free University of Brussels (VUB), Brussels, Belgium [2] Center for Molecular & Vascular Biology, Department of Cardiovascular Sciences, University of Leuven, Leuven, Belgium.
authors.forename: Ermira
authors.initials: E
authors.lastname: Samara-Kuko
authors.affiliation: Vesalius Research Center, Flanders Institute of Biotechnology (VIB) & University of Leuven, Leuven, Belgium.
authors.forename: Abel
authors.initials: A
authors.lastname: Acosta-Sanchez
authors.affiliation: School of Biological Sciences, Royal Holloway - University of London, Egham, UK.
authors.forename: Takis
authors.initials: T
authors.lastname: Athanasopoulos
authors.affiliation: School of Biological Sciences, Royal Holloway - University of London, Egham, UK.
authors.forename: George
authors.initials: G
authors.lastname: Dickson
authors.affiliation: Nuclear Medicine Department, UZ Brussel & In vivo Cellular and Molecular Imaging Lab, Free University of Brussels (VUB), Brussels, Belgium.
authors.forename: Tony
authors.initials: T
authors.lastname: Lahoutte
authors.affiliation: Inflammation Research Center, Flanders Institute of Biotechnology (VIB) and Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium.
authors.forename: Pieter
authors.initials: P
authors.lastname: De Bleser
authors.affiliation: 1] Department of Gene Therapy & Regenerative Medicine, Free University of Brussels (VUB), Brussels, Belgium [2] Center for Molecular & Vascular Biology, Department of Cardiovascular Sciences, University of Leuven, Leuven, Belgium.
authors.forename: Thierry
authors.initials: T
authors.lastname: VandenDriessche
authors.affiliation: 1] Department of Gene Therapy & Regenerative Medicine, Free University of Brussels (VUB), Brussels, Belgium [2] Center for Molecular & Vascular Biology, Department of Cardiovascular Sciences, University of Leuven, Leuven, Belgium.
authors.forename: Marinee K
authors.initials: MK
authors.lastname: Chuah
mesh_terms: D000818:Animals; D001665:Binding Sites; D002318:Cardiovascular Diseases / Q000235:genetics / Q000378:metabolism / Q000473:pathology / Q000628:therapy; D019295:Computational Biology; D003587:Cytomegalovirus / Q000737:chemistry / Q000235:genetics; D000229:Dependovirus / Q000235:genetics*; D015870:Gene Expression; D005818:Genetic Engineering / Q000379:methods; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D016678:Genome; D006801:Humans; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D009206:Myocardium / Q000378:metabolism* / Q000473:pathology; D032383:Myocytes, Cardiac / Q000378:metabolism* / Q000473:pathology; D059372:Nucleotide Motifs; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D014158:Transcription, Genetic; D024723:Ventricular Myosins / Q000235:genetics* / Q000378:metabolism
chemical_list: D014157:Transcription Factors; D024723:Ventricular Myosins