Application of array-Comparative Genomic Hybridization analysis in immune-virotherapy approach

AUTORI

Andrea Vitale^1,3, Cristian Capasso², Eleonora Leggiero⁴, Mariangela Garofalo², Lukasz Kuryk L², Mari Hirvinen², Francesca D’Alessio⁴, Carla Perrotta^3,4, Francesco Verdesca^3,4, Annaluisa Ranieri^3,4, Francesco Salvatore^3,4, Lucio Pastore^3,4, Pasqualina Buono^1,4, Vincenzo Cerullo², Barbara Lombardo^3,4
¹Dipartimento di Scienze Motorie e del Benessere (DiSMeB), Università Parthenope, Via Medina 40, 80133 Napoli, Italy
²Laboratory of ImmunoViroTherapy, Drug Research Program, University of Helsinki, Finland
³Dipartimento di Medicina Molecolare e Biotecnologie Mediche, Università Federico II, Napoli, Italy
⁴CEINGE-Biotecnologie Avanzate, Via G. Salvatore 486, 80145 Napoli, Italy

ABSTRACT

Introduction: oncololytic adenoviruses (OAds), viruses constructed to replicate in tumor cells, improve the outcome of cancer therapy in some cases, such as sarcomas. However, the molecular heterogeneity of tumors requires specific and personalized cancer treatments in order to set up more adequate and effective therapies.
Methods: by using the array Comparative Genomic Hybridization (array-CGH), a molecular approach method, we aimed to identify chromosomal aberrations or Copy Number Variants (CNVs) in three different tumor cell lines: HCT116, SW872 and A2058 selected for their Coxsackievirus and Adenovirus receptor (CAR) expression profile.
Results: the cells showed several duplications of genes involved in replicative Adenovirus cycle (binding, internalization, escape) in the core transport, and in the escape of the viral DNA from the capsid.
Conclusion: in this study, our aim was to identify chromosomal alterations in genes involved in the OAd replication cycle process. Array-CGH method could be useful to design a platform for a screening analysis in order to identify mutations that can contribute to oncolytic virotherapy approach generating a personalized strategy for tumor suppression.

BIBLIOGRAFIA

1. Pennisi E. Training viruses to attack cancers. Science 1998;282:1244-6.
2. Lyng H, Beigi M, Svendsrud DH, et al. Intratumor chromosomal heterogeneity in advanced carcinomas of the uterine cervix. Int J Cancer 2004;111:358-66.
3. Sharma A, Tandon M, Bangari DS, et al. Adenoviral vector-based strategies for cancer therapy. Curr Drug ther 2009;4:117-38.
4. Hanahan D, Weinberg R. Hallmarks of cancer: The next generation. Cell 2011;144:646-74.
5. Connolly J, Schnitt S, Wang H, et al. Tumor structure and tumor stroma generation. Holland-Frei Cancer Med 6th. Hamilton: Ed BC Decker, 2003.
6. Vosecka T, Musil Z, Vicha A. Detection of chromosome changes by cgh, array-cgh and snp array techniques in tumours. Cesk Patol 2014;50:25-9.
7. Zebisch A, Schulz E, Grosso M, et al. Identification of a novel variant of epsilon-gamma-delta-beta thalassemia highlights limitations of next generation sequencing. Am J Hematol 2015;90:E52-4.
8. Capasso C, Hirvinen M, Garofalo M, et al. Oncolytic adenoviruses coated with MHC-I tumor epitopes increase the antitumor immunity and efficacy against melanoma. Oncoimmunology 2016;5:e1105429.
9. Heise C, Hermiston T, Johnson L, et al. An adenovirus E1A mutant that demonstrates potent and selective systemic anti-tumoral efficacy. Nat Med 2000;6:1134-9.
10. Sanna V, Ceglia C, Tarsitano M, et al. Aberrant F8 gene intron 1 inversion with concomitant duplication and deletion in a severe hemophilia A patient from Southern Italy. J Thromb Haemost 2013;11:195-7.
11. Nunziato M, Starnone F, Lombardo B, et al. Fast detection of a BRCA2 large genomic duplication by next generation sequencing as a single procedure: A case report. Int J Mol Sci 2017;18.
12. Lombardo B, Ceglia C, Tarsitano M, et al. Identification of a deletion in the NDUFS4 gene using array-comparative genomic hybridization in a patient with suspected mitochondrial respiratory disease. Gene 2014;535:376-9.
13. Kanerva A, Wang M, Bauerschmitz GJ, et al. Gene transfer to ovarian cancer versus normal tissues with fiber-modified adenoviruses. Mol Ther 2002;5:695-704.
14. Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative pcr and the 2(-delta delta c(t)) method. Methods 2001;25:402-8.
15. Suh JH, Yoon JS, Kwon JB, et al. Identification of genomic aberrations by array comparative genomic hybridization in patients with aortic dissections. Korean J Thorac Cardiovasc Surg 2011;44:123-30.
16. Leopold PL, Crystal RG. Intracellular trafficking of adenovirus: Many means to many ends. Adv Drug Deliv Rev 2007;59:810-21.
17. Fueyo J, Alemany R, Gomez-Manzano C, et al. Preclinical characterization of the antiglioma activity of a tropism-enhanced adenovirus targeted to the retinoblastoma pathway. J Natl Cancer Inst 2003;95:652-60.
18. Bagheri N, Shiina M, Lauffenburger DA, et al. A dynamical systems model for combinatorial cancer therapy enhances oncolytic adenovirus efficacy by MEK-inhibition. PLoS Comput Biol 2011;7:e1001085.
19. Zhao H, Granberg F, Elfineh L, et al. Strategic attack on host cell gene expression during adenovirus infection. J Virol 2003;77:11006-15.
20. Watanabe H, Wada T, Handa H. Transcription factor E4TF1 contains two subunits with different functions. EMBO J 1990;9:841-7.
21. Halbert DN, Cutt JR, Shenk T. Adenovirus Early Region 4 Encodes Functions Required for Efficient DNA Replication, Late Gene Expression, and Host Cell Shutoff. J Virol 1985;56:250-7.