Indirect intracoronary delivery of adenovirus encoding adenylyl cyclase increases left ventricular contractile function in mice.
We performed indirect intracoronary delivery of adenovirus vectors in mice and explored techniques including hypothermia and pharmacological means to increase cardiac gene transfer. Mice were maintained in a normothermic state or cooled to 25 degrees C. The aorta or both the pulmonary artery and aorta were clamped while a needle was advanced into the left ventricular cavity to deliver adenovirus vectors encoding enhanced green fluorescent protein (EGFP) or murine adenylyl cyclase type VI (AC(VI)) with saline, sodium nitroprusside, acetylcholine, or serotonin. Clamping was maintained for 30 s (normothermia) or 2 min (25 degrees C) after adenovirus administration. Mice were killed 7 or 21 days later, and hearts were examined for EGFP expression. Compared with clamping the aorta alone and with no cooling, gene transfer was increased as follows: 1) 1.3-fold with hypothermia to extend dwell time; 2) 4.5-fold by clamping the aorta and the pulmonary artery; 3) 11.4-fold with nitroprusside administration; 4) 11.8-fold with serotonin addition, and 5) 14.3-fold with acetylcholine delivery. Gene expression remained substantial at 21 days, and no significant inflammatory response was seen. Efficacy of the method was tested by performing gene transfer of adenovirus encoding AC(VI). Fourteen days after gene transfer, hearts isolated from mice that received adenovirus encoding AC(VI) showed increased contractile function. Indirect intracoronary delivery of adenovirus vectors in mice is associated with efficient cardiac gene transfer and increased left ventricular function after AC(VI) gene transfer.
Roth
David M
DM
Veterans Affairs San Diego Healthcare System, 3350 La Jolla Village Dr., San Diego, CA 92161, USA. droth@ucsd.edu
Lai
N Chin
NC
Gao
Mei Hua
MH
Drumm
Jeffery D
JD
Jimenez
Jesus
J
Feramisco
James R
JR
Hammond
H Kirk
HK
D000109:Acetylcholine / Q000494:pharmacology; D000256:Adenoviridae / Q000235:genetics*; D000262:Adenylyl Cyclases / Q000235:genetics*; D000818:Animals; D003331:Coronary Vessels; D005260:Female; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D007036:Hypothermia, Induced; D007202:Indicators and Reagents; D007527:Isoenzymes / Q000235:genetics; D008164:Luminescent Proteins / Q000235:genetics; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D009206:Myocardium / Q000473:pathology; D020030:Nitric Oxide Donors / Q000494:pharmacology; D009599:Nitroprusside / Q000494:pharmacology; D012701:Serotonin / Q000494:pharmacology; D016277:Ventricular Function, Left
D007202:Indicators and Reagents; D007527:Isoenzymes; D008164:Luminescent Proteins; D020030:Nitric Oxide Donors; D049452:Green Fluorescent Proteins; D009599:Nitroprusside; D012701:Serotonin; D000262:Adenylyl Cyclases; D000109:Acetylcholine