Objectives: Oncolytic virotherapy is a therapeutic approach to cancer treatment that utilizes native or genetically modified viruses that selectively replicate in cancer cells while displaying minimal adverse effects in normal healthy cells. To date, a wide variety of viruses have been evaluated for their oncolytic potential, including DNA viruses. Our group previously demonstrated that Caprine herpesvirus type 1 (CpHV 1) is able to induce apoptosis in several normal cell lines. Recently, we have characterized in more detail the intracellular pathway by which CpHV 1 is able to induce apoptosis by analyzing the gene expression response during the apoptotic phase of CpHV 1 infection in a murine neuroblastoma cell line. Thus, the aim of the present research was to investigate the ability of CpHV-1 to replicate, cause cell death and affect cellular viability in a panel of human cancer cells lines. Methods: Human breast adenocarcinoma (MDA-MB-468), Human cervical adenocarcinoma (HeLa), Human osteosarcoma (U2OS), Human prostatic adenocarcinoma (PC3), Human lung carcinoma (A549), Human malignant melanoma (A375) and Chronic Myelogenous Leukemia (K562) cell lines were used. Madin Darby bovine kidney (MDBK) cell line was used as control. Citotoxicity was assayed by MTT test; Apoptosis was assayed by using the ApoToxGlo Triplex assay; The espression of apoptosis-related genes was assayed by Western Blot; Viral replication was assayed by TCID50 method. Results: In a first series of experiments we have analyzed the effect of CpHV1 infection on cell viability by means of the MTT assay at different time post infection and several multiplicity of infection. All cell lines, except K562 cells, showed a marked cytopatic effect, demonstrating an oncolytic potential of CpHV-1 in tested human cancer cells. The reduction of cells viability was associated with significant levels of viral production as assayed by TCID50 after 24 h p.i. for MDBK, 293T, MDA-MD468, A549, U2OS, A375 and after 48 h p.i. for PC3, HeLa and K562 cell lines. Then, we investigated virus induced cytotoxicity, viability, and apoptosis within a single assay well, by using the ApoToxGlo Triplex assay. Analysis of virus infected cells revealed activation of caspase3, a marker of apoptosis at 24 h postinfection. Conlusion: Our findings firstly demonstrate an oncolytic effect of CpHV‐1 in neoplastic cell lines in terms of caspase activation and apoptosis modulation suggesting CpHV‐1 as a potential candidate in oncolytic virotherapy.

Caprine Herpesvirus 1 (CpHV-1) a potential candidate for oncolytic virotherapy / Montagnaro, S.; Iovane, V.; Puzio, M. V.; Ciarcia, R.; Rocco, D.; De Martino, L.; Iovane, G.; Pagnini, U.. - (2017), pp. 427-427. (Intervento presentato al convegno XI International Congress of Bacteriology and Applied Microbiology tenutosi a 01/07/17 nel 28/06/17).

Caprine Herpesvirus 1 (CpHV-1) a potential candidate for oncolytic virotherapy

S. Montagnaro
Conceptualization
;
V. Iovane;M. V. Puzio;R. Ciarcia;L. De Martino;G. Iovane;U. Pagnini
2017

Abstract

Objectives: Oncolytic virotherapy is a therapeutic approach to cancer treatment that utilizes native or genetically modified viruses that selectively replicate in cancer cells while displaying minimal adverse effects in normal healthy cells. To date, a wide variety of viruses have been evaluated for their oncolytic potential, including DNA viruses. Our group previously demonstrated that Caprine herpesvirus type 1 (CpHV 1) is able to induce apoptosis in several normal cell lines. Recently, we have characterized in more detail the intracellular pathway by which CpHV 1 is able to induce apoptosis by analyzing the gene expression response during the apoptotic phase of CpHV 1 infection in a murine neuroblastoma cell line. Thus, the aim of the present research was to investigate the ability of CpHV-1 to replicate, cause cell death and affect cellular viability in a panel of human cancer cells lines. Methods: Human breast adenocarcinoma (MDA-MB-468), Human cervical adenocarcinoma (HeLa), Human osteosarcoma (U2OS), Human prostatic adenocarcinoma (PC3), Human lung carcinoma (A549), Human malignant melanoma (A375) and Chronic Myelogenous Leukemia (K562) cell lines were used. Madin Darby bovine kidney (MDBK) cell line was used as control. Citotoxicity was assayed by MTT test; Apoptosis was assayed by using the ApoToxGlo Triplex assay; The espression of apoptosis-related genes was assayed by Western Blot; Viral replication was assayed by TCID50 method. Results: In a first series of experiments we have analyzed the effect of CpHV1 infection on cell viability by means of the MTT assay at different time post infection and several multiplicity of infection. All cell lines, except K562 cells, showed a marked cytopatic effect, demonstrating an oncolytic potential of CpHV-1 in tested human cancer cells. The reduction of cells viability was associated with significant levels of viral production as assayed by TCID50 after 24 h p.i. for MDBK, 293T, MDA-MD468, A549, U2OS, A375 and after 48 h p.i. for PC3, HeLa and K562 cell lines. Then, we investigated virus induced cytotoxicity, viability, and apoptosis within a single assay well, by using the ApoToxGlo Triplex assay. Analysis of virus infected cells revealed activation of caspase3, a marker of apoptosis at 24 h postinfection. Conlusion: Our findings firstly demonstrate an oncolytic effect of CpHV‐1 in neoplastic cell lines in terms of caspase activation and apoptosis modulation suggesting CpHV‐1 as a potential candidate in oncolytic virotherapy.
2017
Caprine Herpesvirus 1 (CpHV-1) a potential candidate for oncolytic virotherapy / Montagnaro, S.; Iovane, V.; Puzio, M. V.; Ciarcia, R.; Rocco, D.; De Martino, L.; Iovane, G.; Pagnini, U.. - (2017), pp. 427-427. (Intervento presentato al convegno XI International Congress of Bacteriology and Applied Microbiology tenutosi a 01/07/17 nel 28/06/17).
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11588/746102
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