RESUMEN
Replication-competent (oncolytic) adenoviruses (OAV) can be adapted as vectors for the delivery of therapeutic genes, with the aim of extending the antitumor effect beyond direct cytolysis. Transgene expression using these vectors is usually intense but short-lived, and repeated administrations are hampered by the rapid appearance of neutralizing antibodies (NAbs). We have studied the performance of monocytes as cell carriers to improve transgene expression in cancer models established in athymic mice and immunocompetent Syrian hamsters. Human and hamster monocytic cell lines (MonoMac6 and HM-1, respectively) were loaded with replication-competent adenovirus-expressing luciferase. Intravenous administration of these cells caused a modest increase in transgene expression in tumor xenografts, but this effect was virtually lost in hamsters. In contrast, intratumoral administration of HM-1 cells allowed repeated cycles of expression and achieved partial protection from NAbs in preimmunized hamsters bearing pancreatic tumors. To explore the therapeutic potential of this approach, HM-1 cells were loaded with a hypoxia-inducible OAV expressing the immunostimulatory cytokine interleukin-12 (IL-12). Three cycles of treatment achieved a significant antitumor effect in the hamster model, and transgene expression was detected following each administration, in contrast with the rapid neutralization of the free virus. We propose monocytes as carriers for multiple intratumoral administrations of armed OAVs.
Asunto(s)
Adenoviridae/genética , Monocitos/virología , Neoplasias Experimentales/terapia , Viroterapia Oncolítica/métodos , Adenoviridae/patogenicidad , Animales , Cricetinae , Expresión Génica , Humanos , Interleucina-12/genética , Interleucina-12/metabolismo , Mesocricetus , Ratones , Ratones Desnudos , Neoplasias Experimentales/genética , Neoplasias Pancreáticas/genética , Neoplasias Pancreáticas/terapia , Transgenes , Trasplantes/virología , Replicación Viral/genética , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
The use of viruses as therapeutic agents against cancer is an old concept that has had a significant revival in the past two decades, in parallel with advances in methods to modify viral genomes genetically. From the initial stage of proof of concept, the field of virotherapy quickly progressed to the clinical setting, where serious limitations, yet promising opportunities, were identified. After demonstrating good safety profiles in humans, the objective in virotherapy has become to improve the efficacy of oncolytic viruses. Virotherapy approaches include incorporating therapeutic genes, evaluating alternative viruses with stronger oncolytic potential, employing new methods to improve biodistribution, and establishing greater insight into the influence of the immune system on both the success and failure of therapies. This review summarizes the most significant advances in recent years in the design of virotherapy for effective tumor treatment.
Asunto(s)
Neoplasias/terapia , Viroterapia Oncolítica/métodos , Animales , Terapia Genética , Humanos , Virus Oncolíticos/fisiología , Distribución Tisular , Replicación Viral/fisiologíaRESUMEN
PURPOSE: Tumors of the Ewing family are characterized by chromosomal translocations that yield chimeric transcription factors, such as EWS/FLI1, which regulate the expression of specific genes that contribute to the malignant phenotype. In the present study, we show that cholecystokinin (CCK) is a new target of the EWS/FLI1 oncoprotein and assess its functional role in Ewing tumor pathogenesis. EXPERIMENTAL DESIGN: Relevant EWS/FLI1 targets were identified using a combination of cell systems with inducible EWS/FLI1 expression, Ewing tumors and cell lines, microarrays, and RNA interference with doxycycline-inducible small hairpin RNA (shRNA) vectors. A doxycycline-inducible CCK-shRNA vector was stably transfected in A673 and SK-PN-DW Ewing cell lines to assess the role of CCK in cell proliferation and tumor growth. RESULTS: Microarray analysis revealed that CCK was up-regulated by EWS/FLI1 in HeLa cells. CCK was overexpressed in Ewing tumors as compared with other pediatric malignancies such as rhabdomyosarcoma and neuroblastoma, with levels close to those detected in normal tissues expressing the highest levels of CCK. Furthermore, EWS/FLI1 knockdown in A673 and SK-PN-DW Ewing cells using two different doxycycline-inducible EWS/FLI1-specific shRNA vectors down-regulated CCK mRNA expression and diminished the levels of secreted CCK, showing that CCK is a EWS/FLI1 specific target gene in Ewing cells. A doxycycline-inducible CCK-specific shRNA vector successfully down-regulated CCK expression, reduced the levels of secreted CCK in Ewing cell lines, and inhibited cell growth and proliferation in vitro and in vivo. Finally, we show that Ewing cell lines and tumors express CCK receptors and that the growth inhibition produced by CCK silencing can be rescued by culturing the cells with medium containing CCK. CONCLUSIONS: Our data support the hypothesis that CCK acts as an autocrine growth factor stimulating the proliferation of Ewing cells and suggest that therapies targeting CCK could be promising in the treatment of Ewing tumors.