Enhanced adenovirus infection of melanoma cells by fiber-modification: incorporation of RGD peptide or Ad5/3 chimerism.
The incidence of malignant melanoma has been increasing. Unfortunately, advanced melanomas are rarely curable with standard therapy; therefore, new forms of treatment such as gene therapy are needed. The success of gene delivery or oncolysis depends on the nature of the vector. Adenoviral vectors are advantageous for several reasons; however, they are dependent on CAR (coxsackie and adenovirus receptor) which is deficient or heterogeneously expressed on melanoma cells in situ. Correspondingly, transduction of freshly purified melanoma cells has been shown to be minimal or variable. In order to overcome this shortcoming, it is necessary to construct tropism modified adenoviral vectors. With this goal in mind, we generated two tropism modified vectors, Ad5lucRGD which has an RGD motif incorporated into the HI loop of the fiber knob and Ad5/3luc1 which contains the tail and shaft domain of the Ad5 fiber and the knob domain of the Ad3 fiber. Herein we demonstrate that Ad5/3luc1 infects CAR-negative primary melanoma cells 1128 times better than Ad5luc1 and 34 times better than Ad5lucRGD. Furthermore we show that Ad5/3luc1 and Ad5lucRGD infect via a CAR independent route by blocking the CAR receptor. In addition, we show that the infectivity of the cells correlates with the expression of CAR and Ad3 receptors determined by FACS analysis. Therefore, Ad5/3 is very attractive as a potential therapeutic vector for malignant melanoma.
Volk
Andrea L
AL
Division of Human Gene Therapy and Gene Therapy Center, Department of Pathology, University of Alabama at Birmingham, Birmginham, Alabama USA.
Rivera
Angel A
AA
Kanerva
Anna
A
Bauerschmitz
Gerd
G
Dmitriev
Igor
I
Nettelbeck
Dirk M
DM
Curiel
David T
DT
D000260:Adenoviruses, Human / Q000235:genetics* / Q000378:metabolism; D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology; D002678:Chimera / Q000235:genetics*; D062727:Coxsackie and Adenovirus Receptor-Like Membrane Protein; D005434:Flow Cytometry; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D005822:Genetic Vectors; D006801:Humans; D016023:Integrins / Q000378:metabolism; D008545:Melanoma / Q000378:metabolism / Q000821:virology*; D009842:Oligopeptides / Q000235:genetics*; D011991:Receptors, Virus / Q000378:metabolism; D014407:Tumor Cells, Cultured
D000970:Antineoplastic Agents; C568771:CLMP protein, human; D062727:Coxsackie and Adenovirus Receptor-Like Membrane Protein; D016023:Integrins; D009842:Oligopeptides; D011991:Receptors, Virus; C047981:arginyl-glycyl-aspartic acid