DIRECTED EVOLUTION OF AN ONCOLYTIC VESICULAR STOMATITIS VIRUS ADAPTED TO HUMAN MALIGNANT MENINGIOMA CELLS

Yıl 2024, , 24 - 31, 15.02.2024

Burak Gizem Göleş , Hülya Yazıcı , John N. Davıs

https://doi.org/10.26650/JARHS2024-1300891

Öz

Objectives: Recombinantly-engineered versions of the oncolytic virus VSV are currently under clinical investigation for the treatment of several different types of cancer. Here we aim to enhance the cancer-killing oncolytic phenotype of VSV-1’GFP toward human malignant meningioma cells using a directed evolution approach.

Material and Methods: Two independent trials of repeated growth of VSV-1’GFP on cultures of meningioma IOMM-Lee cells were performed. This adaptation procedure allows for the selection of viral mutants that display an enhanced oncolytic phenotype. A fluorescent viral plaque assay was used to measure changes in plaque size indicative of enhanced viral growth on these cancer cells. Sanger sequencing was used to identify the viral mutations responsible.

Results: Adapted VSV-1’GFP from each of the growth trials yielded larger fluorescent plaques than control virus, indicating the emergence of viral mutants with increased growth on these meningioma cells. Plaques from adapted virus were 184%±9% (Trial 1) and 166%±7% (Trial 2) larger than control (n=60; p<0.001; ANOVA). Sequencing determined that adapted virus from Trial 1 harbored 3 mutations: a silent mutation Y178Y in the M gene, an E92K mutation in the G gene, and a K152R mutation in the L gene. Trial 2 yielded 3 mutations in the G gene: N36T, E92K, and E254K.

Conclusion: The E92K mutation of the viral G-protein emerged independently in both growth trials, suggesting that this change may play a role in producing the enlarged-plaque phenotype and enhanced oncolytic propagation in IOMM-Lee cells. Further investigations of the prospect for treating malignant meningiomas using VSV-based oncolytic virotherapy appear warranted and, to the best of our knowledge, the present study appears to be the first directed evolution experiment involving an oncolytic virus adapted to human meningioma cells.

Anahtar Kelimeler

Vesicular stomatitis virus, oncolytic virus, meningioma, mutagenesis

Teşekkür

I would like to express my sincere gratitude and appreciation for Dr. Murat GUNEL and his invaluable contribution to this article.

İNSAN MALIGANT MENİNJİOM HÜCRELERİNE UYUMLANMIŞ BİR ONKOLİTİK VEZİKÜLER STOMATİT VİRÜSÜNÜN YÖNLENDİRİLMİŞ EVRİMİ

Öz

Amaç: Onkolitik virüs VSV’nin rekombinant olarak tasarlanmış çeşitleri birkaç farklı kanser türünün tedavisi için klinik olarak araştırılmaktadır. Bu çalışmada, yönlendirilmiş evrim yaklaşımı kullanarak VSV-1’GFP’ nin insan malign meninjiyom hücrelerine yönelik kanser öldürücü onkolitik fenotipini geliştirmek hedeflenmektedir.

Gereç ve Yöntemler: Meninjiyom IOMM-Lee hücrelerinin kültürleri üzerinde VSV-1’GFP’nin tekrarlanan büyümesine ilişkin iki bağımsız deneme gerçekleştirildi. Bu adaptasyon prosedürü, gelişmiş bir onkolitik fenotip gösteren viral mutantların seçimine olanak tanır. Bu kanser hücrelerinde viral büyümenin arttığını gösteren plak boyutundaki değişiklikleri ölçmek için bir floresan viral plak tahlili kullanıldı. Sorumlu viral mutasyonları tanımlamak için Sanger dizilimi kullanıldı.

Bulgular: Büyüme denemelerinin her birinden uyarlanmış VSV-1’GFP, kontrol virüsünden daha büyük floresan plaklar verdi; bu, meninjiyom hücrelerinde artan büyüme ile viral mutantların ortaya çıktığını gösterir. Uyarlanmış virüsten alınan plaklar kontrolden (n=60; p<0,001; ANOVA) %184±%9 (Deneme 1) ve %166±%7 (Deneme 2) daha büyüktü. Dizileme, Deneme 1’den uyarlanan virüsün 3 mutasyon barındırdığını belirledi: M geninde sessiz bir Y178Y mutasyonu, G geninde bir E92K mutasyonu ve L geninde bir K152R mutasyonu. Deneme 2, G geninde 3 mutasyon ortaya çıkardı: N36T, E92K ve E254K.

Sonuç: Viral G-proteininin E92K mutasyonu, her iki büyüme denemesinde de bağımsız olarak ortaya çıktı; bu değişiklik, IOMM-Lee hücrelerinde genişlemiş plak fenotipinin ve gelişmiş onkolitik yayılımın üretilmesinde rol oynayabileceğini düşündürmektedir. Malign meninjiyomların VSV bazlı onkolitik viroterapi kullanılarak tedavi edilmesi olasılığına ilişkin daha fazla araştırma gerekli görülmektedir ve bilgimiz dâhilinde, mevcut çalışma, insan meninjiyom hücrelerine uyarlanmış bir onkolitik virüsü içeren ilk yönlendirilmiş evrim deneyi olarak literatüre katkı sunmaktadır.

Anahtar Kelimeler

Veziküler stomatit virüs, onkolitik virus, meninjiom, mutagenez

Kaynakça

• Goradel NH, Baker AT, Arashkia A, Ebrahimi N, Ghorghanlu S, Negahdari B. Oncolytic virotherapy: Challenges and solutions. Curr Probl Cancer 2021;45(1):100639. google scholar
• Wollmann G, Ozduman K, van den Pol AN. Oncolytic virus therapy for glioblastoma multiforme: concepts and candidates. Cancer J 2012;18:69-81. google scholar
• Desjardins A, Gromeier M, Herndon JE, Beaubier N, Bolognesi DP, et al. Recurrent glioblastoma treated with recombinant poliovirus. N Engl J Med 2018;379(2):150-61. google scholar
• Dispenzieri A, Tong C, LaPlant B, Lacy MQ, Laumann K, Dingli D, et al. Phase I trial of systemic administration of Edmonston strain of measles virus genetically engineered to express the sodium iodide symporter in patients with recurrent or refractory multiple myeloma. Leukemia 2017;31(12):2791-8. google scholar
• Andtbacka RHI, Collichio F, Harrington KJ, Middleton MR, Downey G, et al. Final analyses of OPTiM: a randomized phase III trial of talimogene laherparepvec versus granulocyte-macrophage colony-stimulating factor in unresectable stage III-IV melanoma. J Immunother Cancer 2019;7(1):145. google scholar
• Zhang T, Hong-Ting Jou T, Hsin J, Wang Z, Huang K, Ye J, et al. Talimogene Laherparepvec (T-VEC): A Review of the Recent Advances in Cancer Therapy. J Clin Med 2023;12(3):1098. google scholar
• Zhang Y, Nagal BM. Immunovirotherapy based on recombinant vesicular stomatitis virus: Where are we? Front Immunol 2022;13:898631. google scholar
• Bishnoi S, Tiwari R, Gupta S, Byrareddy SN, Nayak D. Oncotargeting by vesicular stomatitis virus (VSV): Advances in cancer therapy. Viruses 2018;10(2):90. google scholar
• Rose JK, Whitt MA. Rhabdoviridae: The viruses and their replication. In: Knipe DM & Howley PM, editors. Fields Virology 4th Edition. Philadelphia: Lippincott Williams & Wilkins; 2001. p.1221-44. google scholar
• Ozduman K, Wollmann G, Piepmeier JM, van den Pol AN. Systemic vesicular stomatitis virus selectively destroys multifocal glioma and metastatic carcinoma in brain. J Neurosci 2008;28(8):1882-93. google scholar
• Wollmann G, Rogulin V, Simon I, Rose JK, van den Pol AN. Some attenuated variants of vesicular stomatitis virus show enhanced oncolytic activity against human glioblastoma cells relative to normal brain cells. J Virol 2010;84(3):1563-73. google scholar
• van den Pol AN, Davis JN. Highly-attenuated recombinant vesicular stomatitis virus VSV-12’GFP displays immunogenic and oncolytic activity. J Virol 2013;87(2):1019-34. google scholar
• Wollmann G, Davis JN, Bosenberg MW, van den Pol AN. Vesicular stomatitis virus variants selectively infect and kill human melanomas but not normal melanocytes. J Virol 2013; 87(12):6644-59. google scholar
• van den Pol AN, Zhang X, Lima E, Pitruzzello M, Albayrak N, et al. Lassa-VSV chimeric virus targets and destroys human and mouse ovarian cancer by direct oncolytic action and by initiating an anti-tumor response. Virology 2021;555:45-55. google scholar
• Wollmann G, Tattersall P, van den Pol AN. Targeting human glioblastoma cells: Comparison of nine viruses with oncolytic potential. J Virol 2005;79(10):6005-22. google scholar
• Gao Y, Whitaker-Dowling P, Watkins SC, Griffin JA, Bergman I. Rapid adaptation of a recombinant vesicular stomatitis virus to a targeted cell line. J Virol 2006;80(17):8603-12. google scholar
• Garijo R, Hernandez-Alonso P, Rivas C, Diallo J-S, Sanjuan R. Experimental evolution of an oncolytic vesicular stomatitis virus with increased selectivity for p53-deficient cells. PloS One 2014;9(7):e102365. google scholar
• Seegers SL, Frasier C, Greene S, Nesmelova IV, Grdzelishvili VZ. Experimental evolution generates novel oncolytic vesicular stomatitis viruses with improved replication in virus-resistant pancreatic cancer cells. J Virol 2020;94(3):e01643-19. google scholar
• Ozduman K, Wollmann G, Piepmeier JM. Gene therapy for meningiomas. In: Pamir MN, Black PM, Fahlbusch R, editors. Meningiomas: A comprehensive text. Philadelphia: Saunders-Elsevier; 2010. p.681-90. google scholar
• Moliterno J, Omuro A, editors. Meningiomas: Comprehensive strategies for management. Springer-Nature Switzerland AG;2020. p.35-41. google scholar
• Clark VE, Erson-Omay EZ, Serin A, Yin J, Cotney J, Ozduman K, et al. Genomic analysis of non-NF2 meningiomas reveals mutations in TRAF7, KLF4, AKT1, and SMO. Science 2013399(6123):1077-80. google scholar
• Youngblood MW, Gunel M. Molecular genetics of meningiomas. Handb Clin Neurol 2020;169:101-9. google scholar
• Lee WH, Tu YC, Liu MY. Primary intraosseus malignant meningioma of the skull: Case report. Neurosurgery 1988;23(4):505-8. google scholar
• Lee WH. Characterization of a newly established malignant meningioma cell line of the human brain: IOMM-Lee. Neurosurgery 1990;27(3):389-96. google scholar
• Mei Y, Bi WL, Greenwald NF, Agar NY, Beroukhim R, Dunn GP, et al. Genomic profile of human meningioma cell lines. PloS One 2017;12(5):e0178322. google scholar
• Chandran K, Sullivan NJ, Felbor U, Whelan SP, Cunningham JM. Endosomal proteolysis of the Ebola virus glycoprotein is necessary for infection. Science 2005;308(5728):1643-5. google scholar
• Wollmann G, Drokhlyansky E, Davis JN, Cepko C, van den Pol AN. Lassa-vesicular stomatitis chimeric virus safely destroys brain tumors. J Virol 2015;89(13):6711-24. google scholar
• Dulbecco R, Vogt M. Some problems of animal virology as studied by the plaque technique. Cold Spring Harb Symp Quant Biol 1953;18:273-9. google scholar
• Lawson ND, Stillman EA, Whitt MA, Rose JK. Recombinant vesicular stomatitis viruses from DNA. Proc Natl Acad Sci USA 1995;92(10):4477-81. google scholar
• Gallione CJ, Rose JK. A single amino acid substitution in a hydrophobic domain causes temperature-sensitive cell-surface transport of a mutant viral glycoprotein. J Virol 1985; 54(2):374-82. google scholar
• Duffy S. Why are RNA virus mutation rates so damn high? PloS Biol 2018;16(8):e3000003. google scholar
• Steinhauer DA, Domingo E, Holland JJ. Lack of evidence for proofreading mechanismsassociated with an RNA virus polymerase. Gene 1992;122(2):281-8. google scholar
• Elena SF, Miralles R, Cuevas JM, Turner PE, Moya A. The two faces of mutation: Extinction and adaptation in RNA viruses. IUBMB Life 2000;49(1):5-9. google scholar
• Drake JW, Holland JJ. Mutation rates among RNA viruses. Proc Natl Acad Sci USA 1999; 96(24):13910-3. google scholar
• Davis JN, van den Pol AN. Viral mutagenesis as a means for generating novel proteins. J Virol 2010;84(3):1625-30. google scholar
• Kretzschmar E, Peluso R, Schnell MJ, Whitt MA, Rose JK. Normal replication of vesicular stomatitis virus without C proteins. Virology 1996;216(2):309-16. google scholar
• Akpinar F, Timm A, Yin J. High-throughput single-cell kinetics of virus infections in the presence of defective interfering particles. J Virol 2015;90(3):1599-612. google scholar
• Lyles DS, Rupprecht CE. Rhabdoviridae. In: Knipe DM & Howley PM, editors. Fields Virology 5th Edition. Philadelphia: Lippincott Williams & Wilkins; 2007. p. 1363-408. google scholar
• Lam V, Duca KA, Yin J. Arrested spread of vesicular stomatitis virus infections in vitro depends on interferon-mediated antiviral activity. Biotechnol Bioeng 2005;90(7):793-804. google scholar
• Osada N, Kohara A, Yamaji T, Hirayama N, Kasai F, et al. The genome landscape of the African green monkey kidney-derived Vero cell line. DNA Res 2014;21(6):673-83. google scholar
• Kawamura Y, Hua L, Gurtner A, Wong E, Kiyokawa J, et al. Histone deacetylase inhibitors enhance oncolytic herpes simplex virus therapy for malignant meningioma. Biomed Pharmacother 2022;155:113843. google scholar
• Jeetendra E, Robison CS, Albritton LM, Whitt MA. The membrane-proximal domain of vesicular stomatitis virus G protein functions as a membrane fusion potentiator and can induce hemifusion. J Virol 2002;76(23):12300-11. google scholar
• White JM, Whittaker GR. Fusion of enveloped viruses in endosomes. Traffic 2016;17(6):593-614. google scholar