Вопросы вирусологии. 2020; 65: 49-56
Разработка препарата для онколитической иммунотерапии на основе вируса осповакцины (Vaccinia virus, Orthopoxvirus, Chordopoxvirinae, Poxviridae) против рака молочной железы
https://doi.org/10.36233/0507-4088-2020-65-1-49-56Аннотация
Введение. В настоящее время активно развиваются новые направления в лечении рака, одним из которых является онколитическая иммунотерапия. Этот подход заключается в использовании вирусов в качестве онкоселективных цитолитических агентов, способных стимулировать опухолеспецифический и неспецифический иммунный ответ организма.
Цель работы – получение рекомбинантного вируса осповакцины, содержащего в геноме гены, кодирующие иммуностимулирующие молекулы, и изучение его онколитических и иммуностимулирующих свойств в экспериментах in vitro и in vivo.
Материал и методы. Рекомбинантный вирус осповакцины получен с использованием метода временной доминантной селекции. Цитолитическую эффективность вируса оценивали колориметрическим методом (МТТ-тест). Иммуногенность полиэпитопной конструкции в составе вирусного генома оценивали ex vivo стимуляцией клеток цельной крови иммунизированных мышей линии BALB/c в ответ на антигены с последующим определением уровня цитокинов методом иммуноферментного анализа.
Результаты. Получен рекомбинантный вирус осповакцины L-IVP_oncoB, содержащий ген, кодирующий гранулоцитарно-макрофагальный колониестимулирующий фактор в области гена J2R, который кодирует тимидинкиназу. Кроме того, данный вирус содержит искусственно синтезированную генетическую конструкцию, кодирующую иммуноген, состоящий из эпитопов антигенов, гиперэкспрессируемых в злокачественных клетках при раке молочной железы, встроенную в область гена C11L (кодирует вирусный фактор роста). Показано, что проведённые модификации вирусного генома не оказывают влияния на ростовые характеристики вируса при культивировании на культурах клеток CV-1 и 4647, а также определена цитолитическая эффективность вируса в отношении раковых культур клеток различного генеза. В эксперименте in vivo выявлено, что полиэпитопная конструкция в составе генома L-IVP_oncoB способна инициировать изменение профиля цитокинов. Обсуждение. Полученные данные охарактеризовали L-IVP_oncoB как перспективный цитолитический и иммуностимулирующий агент и показали необходимость дальнейшего изучения его свойств в качестве средства онколитической иммунотерапии.
Заключение. Проведены основные эксперименты по оценке биологических свойств полученного L-IVP_oncoB, которые необходимы для характеризации онколитического вируса.
Список литературы
1. Youlden D.R., Cramb S.M., Dunn N.A., Muller J.M., Pyke C.M., Baade P.D. The descriptive epidemiology of female breast cancer: an international comparison of screening, incidence, survival and mortality. Cancer Epidemiol. 2012; 36(3): 237-48. https://doi.org/10.1016/j.canep.2012.02.007
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5. Fong P.C., Boss D.S., Yap T.A., Tutt A., Wu P., Mergui-Roelvink M., et al. Inhibition of poly(ADP-ribose) polymerase in tumors from BRCA mutation carriers. N. Engl. J. Med. 2009; 361(2): 123-34. https://doi.org/10.1056/NEJMoa0900212
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7. Delgado-Bellido D., Serrano-Saenz S., Fernández-Cortés M., Oliver F.J. Vasculogenic mimicry signaling revisited: focus on nonvascular VE-cadherin. Mol. Cancer. 2017; 16(1): 65. https://doi.org/10.1186/s12943-017-0631-x
8. Economopoulou P., Kaklamani V.G., Siziopikou K. The role of Cancer stem cells in breast Cancer initiation and progression: potential Cancer stem cell-directed therapies. Oncologist. 2012; 17(11): 1394-401. https://doi.org/10.1634/theoncologist.2012-0163
9. Garber K. China approves world’s first oncolytic. J. Natl. Cancer Inst. 2006; 98(5): 298-300. https://doi.org/10.1093/jnci/djj111
10. Jaunalksne I., Brokāne L., Petroška D., Rasa A., Alberts P. ECHO-7 oncolytic virus Rigvir® in an adjuvant setting for stage I uveal melanoma; A retrospective case report. Am. J. Ophthalmol. Case Rep. 2020; 17: 100615. http://doi.org/10.1016/j.ajoc.2020.100615
11. Adam J., Robertson J., Donegan E., Voicechovskaja I. NICE guidance for talimogene laherparepvec for unresectable metastatic melanoma. Lancet Oncol. 2016; 17(11): 1485-6. https://doi.org/10.1016/S1470-2045(16)30489-2
12. Falkner F.G., Moss B. Transient dominant selection of recombinant vaccinia viruses. J. Virol. 1990; 64(6): 3108-11.
13. Cheever M.A., Allison J.P., Ferris A.S., Finn O.J., Hastings B.M., Hecht T.T., et al. The Prioritization of Cancer Antigens: A National Cancer Institute Pilot Project for the Acceleration of Translational Research. Clin. Cancer Res. 2009; 15(17): 5323-37. https://doi.org/10.1158/1078-0432.CCR-09-0737
14. Schlom J. Therapeutic Cancer Vaccines: Current Status and Moving Forward. J. Natl. Cancer Inst. 2012; 104(8): 599-613. https://doi.org/10.1093/jnci/djs033
15. Milani A., Sangiolo D., Aglietta M., Valabrega G. Recent advances in the development of breast cancer vaccines. Breast Cancer (Dove Med. Press). 2014; 6: 159-68. https://doi.org/10.2147/BCTT.S38428
16. Thomson S.A., Khanna R., Gardner J., Burrows S.R., Coupar B., Moss D.J., et al. Minimal epitopes expressed in a recombinant polyepitope protein are processed and presented to CD8+ cytotoxic T cells: implications for vaccine design. Proc. Natl. Acad. Sci. USA. 1995; 92(13): 5845-9. https://doi.org/10.1073/pnas.92.13.5845
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18. Seyed N., Taheri T., Vauchy C., Dosset M., Godet Y., Eslamifar A., et al. Immunogenicity Evaluation of a Rationally Designed Polytope Construct Encoding HLA-A*0201 Restricted Epitopes Derived from Leishmania major Related Proteins in HLA-A2/DR1 Transgenic Mice: Steps toward Polytope Vaccine. PLoS One. 2014; 9(10): e108848. https://doi.org/10.1371/journal.pone.0108848
19. Nemec A.A., Wallace S.S., Sweasy J.B. Variant base excision repair proteins: Contributors to genomic instability. Semin. Cancer Biol. 2010; 20(5): 320-8. https://doi.org/10.1016/j.semcancer.2010.10.010
20. Horii R., Akiyama F., Kasumi F., Koike M., Sakamoto G. Spontaneous healing of breast cancer. Breast Cancer. 2005; 12(2): 140-4. https://doi.org/10.2325/jbcs.12.140
21. Allegrezza M.J., Conejo-Garcia J.R. Targeted Therapy and Immunosuppression in the Tumor Microenvironment. Trends Cancer. 2017; 3(1): 19-27. https://doi.org/10.1016/j.trecan.2016.11.009
22. Peres L.P., da Luz F.A., Pultz B.A., Brígido P.C., de Araújo R.A., Goulart L.R. Peptide vaccines in breast cancer: The immunological basis for clinical response. Biotechnol. Adv. 2015; 33(8): 1868-77. https://doi.org/10.1016/j.biotechadv.2015.10.013
23. Borsig L., Wolf M.J., Roblek M., Lorentzen A., Heikenwalder M. Inflammatory chemokines and metastasis — tracing the accessory. Oncogene. 2013; 33(25): 3217-24. https://doi.org/10.1038/onc.2013.272
24. Salazar-Onfray F., López M.N., Mendoza-Naranjo A. Paradoxical effects of cytokines in tumor immune surveillance and tumor immune escape. Cytokine Growth Factor Rev. 2007; 18(1-2): 171-82. https://doi.org/10.1016/j.cytogfr.2007.01.015
25. Chen Q., Daniel V., Maher D.W., Hersey P. Production of IL-10 by melanoma cells: examination of its role in immunosuppression mediated by melanoma. Int. J. Cancer. 1994; 56(5): 755-60. https://doi.org/10.1002/ijc.2910560524
26. Giovarelli M., Musiani P., Modesti A., Dellabona P., Casorati G., Allione A., et al. Local release of IL-10 by transfected mouse mammary adenocarcinoma cells does not suppress but enhances antitumor reaction and elicits a strong cytotoxic lymphocyte and antibody-dependent immune memory. J. Immunol. 1995; 155(6): 3112-23.
27. Chen W., Zlotnik A. IL-10: a novel cytotoxic T cell differentiation factor. J. Immunol. 1991; 147(2): 528-34.
28. Kaufman H.L., Rao J.B., Irivine K.R., Bronte V., Rosenberg S.A., Restifo N.P. Interleukin-10 enhances the therapeutic effectiveness of a recombinant poxvirus-based vaccine in an experimental murine tumor model. J. Immunother. 1999; 22(6): 489-96. https://doi.org/10.1097/00002371-199911000-00003
Problems of Virology. 2020; 65: 49-56
Development of the drug oncolytic immunotherapy based on vaccinia viruses (Vaccinia virus, Orthopoxvirus, Chordopoxvirinae, Poxviridae) against breast cancer
https://doi.org/10.36233/0507-4088-2020-65-1-49-56Abstract
Introduction. Currently, new directions in cancer therapy are actively developing, one of which is oncolytic immunotherapy. This approach would be to use of viruses as cancer specific cytolytic agents capable of stimulating both the tumor-specific and non-specific immune response.
The objective paper was obtain a recombinant vaccinia virus containing genes encoding immunostimulating molecules and study oncolytic and immunostimulating properties of recombinant virus.
Material and methods. MTT test, ELISA, methods of transient dominant selection.
Results. The recombinant vaccinia virus (L-IVP_oncoB) were obtained with deletion of the gene encoding thymidine kinase and had an integrated gene encoding GM-CSF. Also the virus have deletion of the gene encoding viral growth factor and integrated genes encoding synthetic tumor-specific polyepitopic immunogens. It was shown that the modifications made to the viral genome did not affect the growth characteristics of the virus when cultured on CV-1 and 4647 cell cultures, and the cytopathogenic efficacy of the virus was determined in relation to cancer cultures of cells of various genesis. In in vivo experiment, it was revealed that the polyepitopic construct in the genome L-IVP_oncoB is able to initiate a change in the profile of cytokines.
Discussion. The obtained data characterized L-IVP_oncoB as a promising cytopathogenic and immunostimulating agent and showed the need for further study of its properties as means of oncolytic immunotherapy. Conclusion. The basic experiments on the evaluation of the biological properties of the obtained L-IVP_oncoB, which are necessary for the characterization of the oncolytic virus, have been carried out.
References
1. Youlden D.R., Cramb S.M., Dunn N.A., Muller J.M., Pyke C.M., Baade P.D. The descriptive epidemiology of female breast cancer: an international comparison of screening, incidence, survival and mortality. Cancer Epidemiol. 2012; 36(3): 237-48. https://doi.org/10.1016/j.canep.2012.02.007
2. Siegel R.L., Miller K.D., Jemal A. Cancer statistics, 2017. CA Cancer J. Clin. 2017; 67(1): 7-30. https://doi.org/10.3322/caac.21387
3. Rouzier R., Perou C.M., Symmans W.F., Ibrahim N., Cristofanilli M., Anderson K., et al. Breast cancer molecular subtypes respond differently to preoperative chemotherapy. Clin. Cancer Res. 2005; 11(16): 5678-85. https://doi.org/10.1158/1078-0432.CCR-04-2421
4. Nixon N.A., Hannouf M.B., Verma S. A review of the value of human epidermal growth factor receptor 2 (HER2)-targeted therapies in breast cancer. Euro. J. Cancer. 2018; 89: 72-81. https://doi.org/10.1016/j.ejca.2017.10.037
5. Fong P.C., Boss D.S., Yap T.A., Tutt A., Wu P., Mergui-Roelvink M., et al. Inhibition of poly(ADP-ribose) polymerase in tumors from BRCA mutation carriers. N. Engl. J. Med. 2009; 361(2): 123-34. https://doi.org/10.1056/NEJMoa0900212
6. Butti R., Gunasekaran V.P., Kumar T.V.S., Banerjee P., Kundu G.C. Breast cancer stem cells: Biology and therapeutic implications. Int. J. Biochem. Cell. Biol. 2019; 107: 38-52. https://doi.org/10.1016/j.biocel.2018
7. Delgado-Bellido D., Serrano-Saenz S., Fernández-Cortés M., Oliver F.J. Vasculogenic mimicry signaling revisited: focus on nonvascular VE-cadherin. Mol. Cancer. 2017; 16(1): 65. https://doi.org/10.1186/s12943-017-0631-x
8. Economopoulou P., Kaklamani V.G., Siziopikou K. The role of Cancer stem cells in breast Cancer initiation and progression: potential Cancer stem cell-directed therapies. Oncologist. 2012; 17(11): 1394-401. https://doi.org/10.1634/theoncologist.2012-0163
9. Garber K. China approves world’s first oncolytic. J. Natl. Cancer Inst. 2006; 98(5): 298-300. https://doi.org/10.1093/jnci/djj111
10. Jaunalksne I., Brokāne L., Petroška D., Rasa A., Alberts P. ECHO-7 oncolytic virus Rigvir® in an adjuvant setting for stage I uveal melanoma; A retrospective case report. Am. J. Ophthalmol. Case Rep. 2020; 17: 100615. http://doi.org/10.1016/j.ajoc.2020.100615
11. Adam J., Robertson J., Donegan E., Voicechovskaja I. NICE guidance for talimogene laherparepvec for unresectable metastatic melanoma. Lancet Oncol. 2016; 17(11): 1485-6. https://doi.org/10.1016/S1470-2045(16)30489-2
12. Falkner F.G., Moss B. Transient dominant selection of recombinant vaccinia viruses. J. Virol. 1990; 64(6): 3108-11.
13. Cheever M.A., Allison J.P., Ferris A.S., Finn O.J., Hastings B.M., Hecht T.T., et al. The Prioritization of Cancer Antigens: A National Cancer Institute Pilot Project for the Acceleration of Translational Research. Clin. Cancer Res. 2009; 15(17): 5323-37. https://doi.org/10.1158/1078-0432.CCR-09-0737
14. Schlom J. Therapeutic Cancer Vaccines: Current Status and Moving Forward. J. Natl. Cancer Inst. 2012; 104(8): 599-613. https://doi.org/10.1093/jnci/djs033
15. Milani A., Sangiolo D., Aglietta M., Valabrega G. Recent advances in the development of breast cancer vaccines. Breast Cancer (Dove Med. Press). 2014; 6: 159-68. https://doi.org/10.2147/BCTT.S38428
16. Thomson S.A., Khanna R., Gardner J., Burrows S.R., Coupar B., Moss D.J., et al. Minimal epitopes expressed in a recombinant polyepitope protein are processed and presented to CD8+ cytotoxic T cells: implications for vaccine design. Proc. Natl. Acad. Sci. USA. 1995; 92(13): 5845-9. https://doi.org/10.1073/pnas.92.13.5845
17. Eslami N.S., Shokrgozar M.A., Mousavi A., Azadmanesh K., Nomani A., Apostolopoulos V., et al. Simultaneous immunisation with a Wilms’ tumour 1 epitope and its ubiquitin fusions results in enhanced cell mediated immunity and tumour rejection in C57BL/6 mice. Mol. Immunol. 2012; 51(3-4): 325-31. https://doi.org/10.1016/j.molimm.2012.03.033
18. Seyed N., Taheri T., Vauchy C., Dosset M., Godet Y., Eslamifar A., et al. Immunogenicity Evaluation of a Rationally Designed Polytope Construct Encoding HLA-A*0201 Restricted Epitopes Derived from Leishmania major Related Proteins in HLA-A2/DR1 Transgenic Mice: Steps toward Polytope Vaccine. PLoS One. 2014; 9(10): e108848. https://doi.org/10.1371/journal.pone.0108848
19. Nemec A.A., Wallace S.S., Sweasy J.B. Variant base excision repair proteins: Contributors to genomic instability. Semin. Cancer Biol. 2010; 20(5): 320-8. https://doi.org/10.1016/j.semcancer.2010.10.010
20. Horii R., Akiyama F., Kasumi F., Koike M., Sakamoto G. Spontaneous healing of breast cancer. Breast Cancer. 2005; 12(2): 140-4. https://doi.org/10.2325/jbcs.12.140
21. Allegrezza M.J., Conejo-Garcia J.R. Targeted Therapy and Immunosuppression in the Tumor Microenvironment. Trends Cancer. 2017; 3(1): 19-27. https://doi.org/10.1016/j.trecan.2016.11.009
22. Peres L.P., da Luz F.A., Pultz B.A., Brígido P.C., de Araújo R.A., Goulart L.R. Peptide vaccines in breast cancer: The immunological basis for clinical response. Biotechnol. Adv. 2015; 33(8): 1868-77. https://doi.org/10.1016/j.biotechadv.2015.10.013
23. Borsig L., Wolf M.J., Roblek M., Lorentzen A., Heikenwalder M. Inflammatory chemokines and metastasis — tracing the accessory. Oncogene. 2013; 33(25): 3217-24. https://doi.org/10.1038/onc.2013.272
24. Salazar-Onfray F., López M.N., Mendoza-Naranjo A. Paradoxical effects of cytokines in tumor immune surveillance and tumor immune escape. Cytokine Growth Factor Rev. 2007; 18(1-2): 171-82. https://doi.org/10.1016/j.cytogfr.2007.01.015
25. Chen Q., Daniel V., Maher D.W., Hersey P. Production of IL-10 by melanoma cells: examination of its role in immunosuppression mediated by melanoma. Int. J. Cancer. 1994; 56(5): 755-60. https://doi.org/10.1002/ijc.2910560524
26. Giovarelli M., Musiani P., Modesti A., Dellabona P., Casorati G., Allione A., et al. Local release of IL-10 by transfected mouse mammary adenocarcinoma cells does not suppress but enhances antitumor reaction and elicits a strong cytotoxic lymphocyte and antibody-dependent immune memory. J. Immunol. 1995; 155(6): 3112-23.
27. Chen W., Zlotnik A. IL-10: a novel cytotoxic T cell differentiation factor. J. Immunol. 1991; 147(2): 528-34.
28. Kaufman H.L., Rao J.B., Irivine K.R., Bronte V., Rosenberg S.A., Restifo N.P. Interleukin-10 enhances the therapeutic effectiveness of a recombinant poxvirus-based vaccine in an experimental murine tumor model. J. Immunother. 1999; 22(6): 489-96. https://doi.org/10.1097/00002371-199911000-00003
