After 6 years of follow-up treating 364 canine melanoma patients, we present here results about the proof-of-concept, safety, and efficacy of a new surgery adjuvant combined gene therapy. The adjuvant treatment (AT) group was divided in three arms as follows: (i) complete surgery plus vaccine (CS-V), (ii) complete surgery plus combined treatment (CS-CT), and (iii) partial surgery plus combined treatment (PS-CT). Besides the genetic vaccines composed by tumor extracts and lipoplexes carrying human interleukin-2 and granulocyte-macrophage colony-stimulating factor genes, the patients were subjected to combined treatment received in the post-surgical bed injections of lipid-complexed thymidine kinase suicide gene plus ganciclovir and canine interferon-β gene plus bleomycin. As compared with surgery-only treated controls (So), CS-CT and CS-V treatments significantly increased the fraction of local disease-free (from 20 to 89 and 74%) and distant metastases-free patients (M0: from 45 to 87 and 84%). Although less effective than CS arms, PS-CT arm demonstrated a significantly improved control of metastatic disease (M0: 80%) compared with So (M0: 44%). In addition, AT produced a significant 9.3- (CS-CT), 6.5- (CS-V), and 5.4-fold (PS-CT) increase of overall survival as compared with their respective So controls. In general terms, the AT changed a lethal disease into a chronic disease where 70% of CS-CT, 51% of CS-V, and 14% of PS-CT patients died of melanoma unrelated causes. These surgery adjuvant treatments delayed or prevented post-surgical recurrence and distant metastasis, and improved disease-free and overall survival while maintaining quality of life. These successful outcomes encourage assaying a similar scheme for human melanoma.
Access optionsAccess options
Subscribe to Journal
Get full journal access for 1 year
only $68.00 per issue
All prices are NET prices.
VAT will be added later in the checkout.
Rent or Buy article
Get time limited or full article access on ReadCube.
All prices are NET prices.
Publisher’s note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Simpson RM, Bastian BC, Michael HT, Webster JD, Prasad ML, Conway CM, et al. Sporadic naturally occurring melanoma in dogs as a preclinical model for human melanoma. Pigment Cell Melanoma Res. 2014;27:37–47.
Smedley RC, Spangler WL, Esplin DG, Kitchell BE, Bergman PJ, Ho HY, et al. Prognostic markers for canine melanocytic neoplasms: a comparative review of the literature and goals for future investigation. Vet Pathol. 2011;48:54–72.
Boston SE, Lu X, Culp WT, Montinaro V, Romanelli G, Dudley RM, et al. Efficacy of systemic adjuvant therapies administered to dogs after excision of oral malignant melanomas: 151 cases (2001-2012). J Am Vet Med Assoc. 2014;245:401–7.
Tuohy JL, Selmic LE, Worley DR, Ehrhart NP, Withrow SJ. Outcome following curative-intent surgery for oral melanoma in dogs: 70 cases (1998-2011). J Am Vet Med Assoc. 2014;245:1266–73.
Hansen K, Khanna C. Spontaneous and genetically engineered animal models; use in preclinical cancer drug development. Eur J Cancer. 2004;40:858–80.
Atherton MJ, Morris JS, McDermott MR, Lichty BD. Cancer immunology and canine malignant melanoma: a comparative review. Vet Immunol Immunopathol. 2016;169:15–26.
Bergman PJ. Veterinary oncology immunotherapies. Vet Clin North Am Small Anim Pract. 2018;48:257–77.
Glikin GC, Finocchiaro LM. Clinical trials of immunogene therapy for spontaneous tumors in companion animals. Sci World J. 2014;2014:718520.
Finocchiaro LME, Glikin GC. Recent clinical trials of cancer immunogene therapy in companion animals. World J Exp Med. 2017;7:42–8.
Quintin-Colonna F, Devauchelle P, Fradelizi D, Mourot B, Faure T, Kourilsky P, et al. Gene therapy of spontaneous canine melanoma and feline fibrosarcoma by intratumoral administration of histoincompatible cells expressing human interleukin-2. Gene Ther. 1996;3:1104–12.
Hogge GS, Burkholder JK, Culp J, Albertini MR, Dubielzig RR, Keller ET, et al. Development of human granulocyte-macrophage colony-stimulating factor-transfected tumor cell vaccines for the treatment of spontaneous canine cancer. Hum Gene Ther. 1998;9:1851–61.
Alexander AN, Huelsmeyer MK, Mitzey A, Dubielzig RR, Kurzman ID, Macewen EG, et al. Development of an allogeneic whole-cell tumor vaccine expressing xenogeneic gp100 and its implementation in a phase II clinical trial in canine patients with malignant melanoma. Cancer Immunol Immunother. 2006;55:433–42.
Dow SW, Elmslie RE, Willson AP, Roche L, Gorman C, Potter TA. In vivo tumor transfection with superantigen plus cytokine genes induces tumor regression and prolongs survival in dogs with malignant melanoma. J Clin Invest. 1998;101:2406–14.
Bergman PJ, McKnight J, Novosad A, Charney S, Farrelly J, Craft D, et al. Long-term survival of dogs with advanced malignant melanoma after DNA vaccination with xenogeneic human tyrosinase: a phase I trial. Clin Cancer Res. 2003;9:1284–90.
Manley CA, Leibman NF, Wolchok JD, Rivière IC, Bartido S, Craft DM, et al. Xenogeneic murine tyrosinase DNA vaccine for malignant melanoma of the digit of dogs. J Vet Intern Med. 2011;25:94–9.
Bianco SR, Sun J, Fosmire SP, Hance K, Padilla ML, Ritt MG, et al. Enhancing antimelanoma immune responses through apoptosis. Cancer Gene Ther. 2003;10:726–36.
Reed SD, Fulmer A, Buckholz J, Zhang B, Cutrera J, Shiomitsu K, et al. Bleomycin/interleukin-12 electrochemogene therapy for treating naturally occurring spontaneous neoplasms in dogs. Cancer Gene Ther. 2010;17:457–64.
Milevoj N, Tratar UL, Nemec A, Brožič A, Žnidar K, Serša G, et al. A combination of electrochemotherapy, gene electrotransfer of plasmid encoding canine IL-12 and cytoreductive surgery in the treatment of canine oral malignant melanoma. Res Vet Sci. 2018;122:40–9.
Westberg S, Sadeghi A, Svensson E, Segall T, Dimopoulou M, Korsgren O, et al. Treatment efficacy and immune stimulation by AdCD40L gene therapy of spontaneous canine malignant melanoma. J Immunother. 2013;36:350–8.
Piras LA, Riccardo F, Iussich S, Maniscalco L, Gattino F, Martano M, et al. Prolongation of survival of dogs with oral malignant melanoma treated by en bloc surgical resection and adjuvant CSPG4-antigen electrovaccination. Vet Comp Oncol. 2017;15:996–1013.
Finocchiaro LM, Fiszman GL, Karara AL, Glikin GC. Suicide gene and cytokines combined nonviral gene therapy for spontaneous canine melanoma. Cancer Gene Ther. 2008;15:165–72.
Finocchiaro LM, Glikin GC. Cytokine-enhanced vaccine and suicide gene therapy as surgery adjuvant treatments for spontaneous canine melanoma. Gene Ther. 2008;15:267–76.
Finocchiaro LM, Glikin GC. Cytokine-enhanced vaccine and suicide gene therapy as surgery adjuvant treatments for spontaneous canine melanoma: 9 years of follow-up. Cancer Gene Ther. 2012;19:852–61.
Finocchiaro LM, Fondello C, Gil-Cardeza ML, Rossi Ú, Villaverde MS, Riveros MD, et al. Cytokine-enhanced vaccine and interferon-β plus suicide gene therapy as surgery adjuvant treatments for spontaneous canine melanoma. Hum Gene Ther. 2015;26:367–76.
Villaverde MS, Gil-Cardeza ML, Glikin GC, Finocchiaro LM. Interferon-β lipofection I. Increased efficacy of chemotherapeutic drugs on human tumor cells derived monolayers and spheroids. Cancer Gene Ther. 2012;19:508–16.
Fondello C, Agnetti L, Villaverde MS, Simian M, Glikin GC, Finocchiaro LME. The combination of bleomycin with suicide or interferon-β gene transfer is able to efficiently eliminate human melanoma tumor initiating cells. Biomed Pharmacother. 2016;83:290–301.
Agnetti L, Fondello C, Villaverde MS, Glikin GC, Finocchiaro LME. Therapeutic potential of bleomycin plus suicide or interferon-β gene transfer combination for spontaneous feline and canine melanoma. Oncoscience. 2017;4:199–214.
Gil-Cardeza ML, Villaverde MS, Fiszman GL, Altamirano NA, Cwirenbaum RA, Glikin GC, et al. Suicide gene therapy on spontaneous canine melanoma: correlations between in vivo tumors and their derived multicell spheroids in vitro. Gene Ther. 2010;17:26–36.
Gil-Cardeza ML, Rossi Ú, Villaverde MS, Glikin GC, Finocchiaro LM. Cationic lipid:DNA complexes allow bleomycin uptake by melanoma cells. Biomed Pharmacother. 2013;67:269–75.
Fondello C, Agnetti L, Glikin GC, Finocchiaro LME. Mechanisms enhancing the cytotoxic effects of bleomycin plus suicide or interferon-β gene lipofection in metastatic human melanoma cells. Anticancer Agents Med Chem. 2018;18:1338–48.
Veterinary Co-operative Oncology Group (VCOG). Veterinary Co-operative Oncology Group - Common Terminology Criteria for Adverse Events (VCOG-CTCAE) following chemotherapy or biological antineoplastic therapy in dogs and cats v1.0. Vet Comp Oncol. 2004;2:195–213.
Villaverde MS, Combe K, Duchene AG, Wei MX, Glikin GC, Finocchiaro LM. Suicide plus immune gene therapy prevents post-surgical local relapse and increases overall survival in an aggressive mouse melanoma setting. Int Immunopharmacol. 2014;22:167–75.
Finocchiaro LME, Spector AIM, Agnetti L, Arbe MF, Glikin GC. Combination of suicide and cytokine gene therapies as surgery adjuvant for canine mammary carcinoma. Vet Sci. 2018;5:70.
Thamm DH, Kurzman ID, Clark MA, Ehrhart EJ 3rd, Kraft SL, Gustafson DL, et al. Preclinical investigation of PEGylated tumor necrosis factor alpha in dogs with spontaneous tumors: phase I evaluation. Clin Cancer Res. 2010;16:1498–508.
Moher D, Hopewell S, Schulz KF, Montori V, Gøtzsche PC, Devereaux PJ, et al. CONSORT 2010 explanation and elaboration: updated guidelines for reporting parallel group randomised trials. BMJ. 2010;340:c869.
We are grateful to our patients and their owners for their cooperation and participation in this study. We recognize the technical assistance and advice of MSc. Doris Riveros, Ms. Graciela Zenobi, and MSc. Juan Cardini. We thank all VMDs involved in this study for patients’ treatment and care, especially Drs. Fernando Calcagno, José L. Suárez, Pablo Meyer, Julián Piñeyra, Jorge Blomberg, Soledad Ramírez, Agustina Spector, Alexis Jalikias, Marie Maminska, Lorena Peteta, Martín Aureggi, and Alejandro Goldman. This work was partially supported by grants from ANPCYT/FONCYT (PICT2012-1738 and PICT2014-1652) and CONICET (PIP 11220110100627 and PIP 11220150100885). L.M.E.F. and G.C.G. are investigators of the Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET, Argentina), and C.F. and L.A. are research fellows of the CONICET.