Standard heparin, low molecular weight heparin, low molecular weight heparinoid, and recombinant hirudin differ in their ability to inhibit transduction by recombinant adeno-associated virus type 2 vectors.
Recombinant adeno-associated virus type 2 (rAAV) is a promising vector for in vivo gene therapy. Transduction by rAAV requires binding to heparan sulfate proteoglycan on the cell surface, and heparin can block this binding. Because heparin is administered to most patients undergoing cardiovascular gene transfer in order to prevent thrombotic events, it is important to identify anticoagulants which do not interfere with rAAV transduction. Therefore, we examined the influence of different anticoagulants on rAAV transduction in vitro. rAAV transduction was inhibited by 40.5 +/- 7.9% at heparin concentrations of 0.1 U/ml, and by 81.7 +/- 3.6% at 1.0 U/ml. The low molecular weight (LMW) heparin tinzaparin inhibited rAAV transduction by 20.2 +/- 3.8% at 0.1 U/ml and 37.1 +/- 1.8% at 1.0 U/ml. The inhibitory effect was significantly weaker compared with heparin at 1.0 U/ml, (P < 0.01). The LMW heparinoid danaparoid inhibited rAAV transduction by 8.8 +/- 3.5% at 0.1 U/ml (P < 0.01 compared with heparin). In contrast, recombinant hirudin did not interfere at all with rAAV transduction. In summary, the results demonstrate that inhibition of rAAV transduction by heparin occurs rapidly and at therapeutically used concentrations. LMW heparinoids and above all recombinant hirudin might be alternatives for heparin when vascular gene transfer with rAAV requires transient anticoagulation.
Hacker
U T
UT
Gene Center of the Ludwig-Maximilians-University Munich, Munich, Germany.
Gerner
F M
FM
Büning
H
H
Hutter
M
M
Reichenspurner
H
H
Stangl
M
M
Hallek
M
M
D000925:Anticoagulants / Q000494:pharmacology*; D002809:Chondroitin Sulfates / Q000494:pharmacology; D000229:Dependovirus / Q000235:genetics*; D003871:Dermatan Sulfate / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004338:Drug Combinations; D005822:Genetic Vectors / Q000008:administration & dosage*; D006367:HeLa Cells / Q000187:drug effects*; D006493:Heparin / Q000494:pharmacology*; D006495:Heparin, Low-Molecular-Weight / Q000494:pharmacology; D006497:Heparan Sulfate / Q000494:pharmacology; D006629:Hirudins / Q000031:analogs & derivatives / Q000494:pharmacology; D006801:Humans; D011994:Recombinant Proteins / Q000494:pharmacology; D000078222:Tinzaparin; D014161:Transduction, Genetic
D000925:Anticoagulants; D002809:Chondroitin Sulfates; D003871:Dermatan Sulfate; D004338:Drug Combinations; D006493:Heparin; D006495:Heparin, Low-Molecular-Weight; D006497:Heparan Sulfate; D006629:Hirudins; D011994:Recombinant Proteins; D000078222:Tinzaparin; C035838:danaparoid; C083544:lepirudin