Combined therapy of colon carcinomas with an oncolytic adenovirus and valproic acid: Contribution of confocal microscopy to our understanding of molecular mechanisms
Karim Benihoud, CNRS UMR 8203, Laboratoire de Vectorologie et Thérapeutiques Anticancéreuses, Univ Paris-Sud, Orsay, France.
The anti-tumor potential of oncolytic adenoviruses (CRAds) has been demonstrated in preclinical and clinical studies. While these agents failed to eradicate tumors when used as a monotherapy, they may be more effective if combined with conventional treatments such as radiotherapy or chemotherapy. This study seeks to evaluate the combination of a CRAd bearing a D24 deletion in E1A with valproic acid (VPA), a histone deacetylase inhibitor for the treatment of human colon carcinomas. This combination led to a dramatic inhibition of cell growth both in vitro and in vivo compared to treatment with CRAd or VPA alone. This effect did not stem from a better CRAd replication and production in the presence of VPA. Inhibition of cell proliferation and a non-apoptotic cell death were shown to be two mechanisms mediating the effects of the combined treatment. Moreover, whereas cells treated only with CRAd displayed a polyploidy (> 4N population), this phenotype was strongly increased in cells treated with both CRAd and VPA, as documented by flow cytometry and confocal microscopy studies. In addition, the increase in polyploidy triggered by combined treatment with CRAd and VPA was associated with the enhancement of H2AX phosphorylation (gH2AX), a hallmark of DNA damage. Finally, E1 and/or viral replication were shown to play a key role in the observed effects since no enhancement of polyploidy nor increase in gH2AX were found following cell treatment with a replication-deficient Ad and VPA. Taken together, our results suggest that CRAd and VPA could be used in combination for the treatment of colon carcinomas.
