Effect of vectors on human endothelial cell signal transduction: implications for cardiovascular gene therapy.
OBJECTIVE
Endothelium is an important target for gene therapy. We have investigated the effect of viral and nonviral vectors on the phenotype and function of endothelial cells (ECs) and developed methods to block any activation caused by these vectors.


RESULTS
Transduction of ECs with viral vectors, including adenovirus, lentiviruses, and Moloney murine leukemia virus, can induce a pro-inflammatory phenotype. This activation was reduced when nonviral vectors were used. We demonstrate that after transduction there is upregulation of dsRNA-triggered antiviral and PI3K/Akt signaling pathway. Blockade of the NFkappaB, PI3-K, or PKR signaling pathways all operated to inhibit partially virally induced activation, and inhibition of both PKR and PI3-K pathways totally blocked EC activation. Furthermore, inhibition of IFN-alpha/beta in addition to PI3-K was effective at preventing EC activation.


CONCLUSIONS
Viral vectors, although efficient at transducing ECs, result in their activation. Blockade of the signaling pathways involved in viral activation may be used to prevent such activation.
Tan
Peng H
PH
Division of Medicine, Department of Immunology, Imperial College London, Hammersmith Hospital, London, UK. ptan@imperial.ac.uk
Xue
Shao-An
SA
Manunta
Maria
M
Beutelspacher
Sven C
SC
Fazekasova
Henrieta
H
Alam
A K M Shamsul
AK
McClure
Myra O
MO
George
Andrew J T
AJ
D000256:Adenoviridae / Q000235:genetics; D002318:Cardiovascular Diseases / Q000276:immunology / Q000378:metabolism / Q000628:therapy*; D002448:Cell Adhesion / Q000276:immunology; D002465:Cell Movement / Q000276:immunology; D002478:Cells, Cultured; D016207:Cytokines / Q000037:antagonists & inhibitors / Q000276:immunology; D004730:Endothelium, Vascular / Q000166:cytology / Q000276:immunology / Q000378:metabolism*; D015316:Genetic Therapy / Q000009:adverse effects* / Q000379:methods; D005822:Genetic Vectors / Q000276:immunology*; D006801:Humans; D016086:Lentivirus / Q000235:genetics; D008979:Moloney murine leukemia virus / Q000235:genetics; D010641:Phenotype; D019869:Phosphatidylinositol 3-Kinases / Q000009:adverse effects / Q000378:metabolism; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D012501:Saphenous Vein / Q000166:cytology; D015398:Signal Transduction / Q000276:immunology*; D018417:Th1 Cells / Q000166:cytology / Q000276:immunology; D014161:Transduction, Genetic; D014657:Vasculitis / Q000209:etiology* / Q000276:immunology / Q000378:metabolism
D016207:Cytokines; D019869:Phosphatidylinositol 3-Kinases; D051057:Proto-Oncogene Proteins c-akt