title: Myocardial gene delivery using molecular cardiac surgery with recombinant adeno-associated virus vectors in vivo.
abstract: We use a novel technique that allows for closed recirculation of vector genomes in the cardiac circulation using cardiopulmonary bypass, referred to here as molecular cardiac surgery with recirculating delivery (MCARD). We demonstrate that this platform technology is highly efficient in isolating the heart from the systemic circulation in vivo. Using MCARD, we compare the relative efficacy of single-stranded (ss) adeno-associated virus (AAV)6, ssAAV9 and self-complimentary (sc)AAV6-encoding enhanced green fluorescent protein, driven by the constitutive cytomegalovirus promoter to transduce the ovine myocardium in situ. MCARD allows for the unprecedented delivery of up to 48 green fluorescent protein genome copies per cell globally in the sheep left ventricular (LV) myocardium. We demonstrate that scAAV6-mediated MCARD delivery results in global, cardiac-specific LV gene expression in the ovine heart and provides for considerably more robust and cardiac-specific gene delivery than other available delivery techniques such as intramuscular injection or intracoronary injection; thus, representing a potential, clinically translatable platform for heart failure gene therapy.
authors.affiliation: Division of Cardiovascular Surgery, Department of Surgery, University of Pennsylvania School of Medicine, Philadelphia, PA 19104, USA.
authors.forename: J D
authors.initials: JD
authors.lastname: White
authors.forename: D M
authors.initials: DM
authors.lastname: Thesier
authors.forename: J B D
authors.initials: JB
authors.lastname: Swain
authors.forename: M G
authors.initials: MG
authors.lastname: Katz
authors.forename: C
authors.initials: C
authors.lastname: Tomasulo
authors.forename: A
authors.initials: A
authors.lastname: Henderson
authors.forename: L
authors.initials: L
authors.lastname: Wang
authors.forename: C
authors.initials: C
authors.lastname: Yarnall
authors.forename: A
authors.initials: A
authors.lastname: Fargnoli
authors.forename: M
authors.initials: M
authors.lastname: Sumaroka
authors.forename: A
authors.initials: A
authors.lastname: Isidro
authors.forename: M
authors.initials: M
authors.lastname: Petrov
authors.forename: D
authors.initials: D
authors.lastname: Holt
authors.forename: R
authors.initials: R
authors.lastname: Nolen-Walston
authors.forename: W J
authors.initials: WJ
authors.lastname: Koch
authors.forename: H H
authors.initials: HH
authors.lastname: Stedman
authors.forename: J
authors.initials: J
authors.lastname: Rabinowitz
authors.forename: C R
authors.initials: CR
authors.lastname: Bridges
mesh_terms: D000818:Animals; D006348:Cardiac Surgical Procedures / Q000379:methods*; D002315:Cardiopulmonary Bypass; D003587:Cytomegalovirus; D000229:Dependovirus / Q000235:genetics*; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000235:genetics; D009206:Myocardium / Q000378:metabolism; D012756:Sheep
chemical_list: D049452:Green Fluorescent Proteins