Conditionally replicative adenovirus with tropism expanded towards integrins inhibits osteosarcoma tumor growth in vitro and in vivo.
OBJECTIVE
The clinical course of osteosarcoma (OS) demands the development of new therapeutic options. Conditionally replicative adenoviruses (CRAds) represent promising agents for the treatment of solid tumors, because CRAds have an intrinsic replication capacity that allows in situ amplification and extensive tumor infection through lateral spread. The CRAd AdDelta24 has been developed to replicate selectively in cells with a defective retinoblastoma (Rb) pathway. Because genetic alterations in the Rb pathway are frequently observed in OS, AdDelta24 might be useful in the treatment of this cancer.


METHODS
Because the lack of Coxsackie adenovirus receptor on OS cells limits the efficacy of CRAd treatment, we explored alternative target molecules on OS. Insertion of an Arg-Gly-Asp (RGD-4C) integrin-targeting motif into the adenovirus fiber knob expanded tropism toward the alpha(nu)beta(3) and alpha(nu)beta(5) integrins. The oncolytic capacity of the CRAd Ad5-Delta24RGD was tested on primary OS cells in vitro and in vivo.


RESULTS
The alpha(nu)beta(3)and alpha(nu)beta(5) integrins are abundantly expressed on OS cells. RGD-mediated infection augmented adenovirus infection of primary OS cells by two orders of magnitude. Ad5-Delta24RGD was shown to be highly active in killing human OS cell lines, as well as primary cell cultures. Furthermore, intratumoral injections with Ad5-Delta24RGD into established human OS xenografts that were derived directly from a patient with OS refractory for chemotherapeutic treatment caused a significant tumor-growth delay. Furthermore, adenoviral particles could be detected in tumor cells 25 days posttumor injection.


CONCLUSIONS
Targeting adenovirus toward integrins rendered OS cells more sensitive to killing by Ad5-Delta24RGD. These findings suggest that Ad5-Delta24RGD has potential for use in OS treatment.
Witlox
Adhiambo M
AM
Department of Orthopedic Surgery, VU University Medical Center, Amsterdam, The Netherlands.
Van Beusechem
Victor W
VW
Molenaar
Bonnie
B
Bras
Hans
H
Schaap
Gerard R
GR
Alemany
Ramon
R
Curiel
David T
DT
Pinedo
Herbert M
HM
Wuisman
Paul I J M
PI
Gerritsen
Winald R
WR
D000256:Adenoviridae / Q000502:physiology*; D000818:Animals; D001859:Bone Neoplasms / Q000378:metabolism / Q000517:prevention & control* / Q000821:virology; D002470:Cell Survival; D005260:Female; D015316:Genetic Therapy; D006801:Humans; D066298:In Vitro Techniques; D039302:Integrin alphaVbeta3 / Q000378:metabolism*; D016023:Integrins / Q000378:metabolism*; D051379:Mice; D008819:Mice, Nude; D009842:Oligopeptides / Q000235:genetics / Q000378:metabolism; D012516:Osteosarcoma / Q000378:metabolism / Q000517:prevention & control* / Q000821:virology; D019030:Receptors, Vitronectin / Q000378:metabolism*; D016160:Retinoblastoma Protein / Q000172:deficiency / Q000235:genetics; D016879:Salvage Therapy; D018523:Tropism; D014407:Tumor Cells, Cultured; D014779:Virus Replication; D023041:Xenograft Model Antitumor Assays
D039302:Integrin alphaVbeta3; D016023:Integrins; D009842:Oligopeptides; D019030:Receptors, Vitronectin; D016160:Retinoblastoma Protein; C064660:integrin alphaVbeta5; C047981:arginyl-glycyl-aspartic acid