{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,11]],"date-time":"2026-09-11T09:07:12Z","timestamp":1789117632829,"version":"build-2803163510"},"reference-count":29,"publisher":"MDPI AG","issue":"18","license":[{"start":{"date-parts":[[2026,9,11]],"date-time":"2026-09-11T00:00:00Z","timestamp":1789084800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100006769","name":"Russian Science Foundation","doi-asserted-by":"publisher","award":["24-14-00390"],"award-info":[{"award-number":["24-14-00390"]}],"id":[{"id":"10.13039\/501100006769","id-type":"DOI","asserted-by":"publisher"}]},{"award":["24-14-00390"],"award-info":[{"award-number":["24-14-00390"]}],"id":[{"id":"https:\/\/ror.org\/03y2gwe85","id-type":"ROR","asserted-by":"publisher"}]},{"name":"Russian state-funded","award":["125012900932-4"],"award-info":[{"award-number":["125012900932-4"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Int. J. Mol. Sci.","IJMS"],"abstract":"<jats:p>Both oncolytic virotherapy and immune checkpoint blockade are being actively explored as immunotherapy treatments for glioblastoma, one of the most lethal malignancies. Nevertheless, the antitumor molecular mechanisms of these therapies remain poorly understood, particularly whether they induce overlapping or distinct transcriptional programs. No comprehensive transcriptome-wide characterization and direct comparison of the responses induced by VV-GMCSF-Lact and anti-PD-1 therapy in glioma have been reported. To address this question, we performed whole-transcriptome profiling of GL261 tumors from immunocompetent C57BL\/6 mice treated with VV-GMCSF-Lact, anti-PD-1 antibodies, or their combination. We analyzed therapy-associated changes in gene expression, signaling pathways, and tumor composition. Our findings indicate that anti-PD-1 and combination treatment led to distinctive activation of interferon-gamma response, JAK\/STAT signaling, and TNF-alpha\/NF-kB pathways. In contrast, VV-GMCSF-Lact preferentially activated T, B, and natural killer T cell-associated programs and reduced the relative abundance of malignant cells. Notably, combination therapy was associated with unique molecular patterns, including induction of monocyte and granulocyte chemotaxis-associated genes, downregulation of PP2A-regulated signaling, and suppression of Rap1-related pathways. This suggests that VV-GMCSF-Lact and anti-PD-1 therapy induce transcriptionally distinct, yet complementary, immune reprogramming and support further evaluation of their combination for glioma immunotherapy.<\/jats:p>","DOI":"10.3390\/ijms27188078","type":"journal-article","created":{"date-parts":[[2026,9,11]],"date-time":"2026-09-11T08:17:42Z","timestamp":1789114662000},"page":"8078","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Whole-Transcriptome Profiling of Murine GL261 Glioma Reveals Signatures Consistent with Distinct Immune Reprogramming Induced by Oncolytic Virotherapy with VV-GMCSF-Lact and Anti-PD-1 Therapy"],"prefix":"10.3390","volume":"27","author":[{"ORCID":"https:\/\/orcid.org\/0009-0000-4033-4724","authenticated-orcid":false,"given":"Anna S.","family":"Chesnokova","sequence":"first","affiliation":[{"name":"Institute of Chemical Biology and Fundamental Medicine, Siberian Branch, Russian Academy of Sciences, Lavrentyev Avenue 8, Novosibirsk 630090, Russia"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"conceptualization","vocabulary":"credit"},{"role":"methodology","vocabulary":"credit"},{"role":"software","vocabulary":"credit"},{"role":"formal-analysis","vocabulary":"credit"},{"role":"investigation","vocabulary":"credit"},{"role":"data-curation","vocabulary":"credit"},{"role":"writing-original-draft","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6909-7301","authenticated-orcid":false,"given":"Alisa B.","family":"Ageenko","sequence":"additional","affiliation":[{"name":"Institute of Chemical Biology and Fundamental Medicine, Siberian Branch, Russian Academy of Sciences, Lavrentyev Avenue 8, Novosibirsk 630090, Russia"},{"name":"Research Center for Genetics and Life Sciences, Sirius University of Science and Technology, Krasnodar Region, Sirius (Federal Territory of Sirius), Sochi 354340, Russia"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"methodology","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0392-794X","authenticated-orcid":false,"given":"Natalia S.","family":"Vasileva","sequence":"additional","affiliation":[{"name":"Institute of Chemical Biology and Fundamental Medicine, Siberian Branch, Russian Academy of Sciences, Lavrentyev Avenue 8, Novosibirsk 630090, Russia"},{"name":"Research Center for Genetics and Life Sciences, Sirius University of Science and Technology, Krasnodar Region, Sirius (Federal Territory of Sirius), Sochi 354340, Russia"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"methodology","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0009-0000-7318-4902","authenticated-orcid":false,"given":"Arina A.","family":"Byvakina","sequence":"additional","affiliation":[{"name":"Institute of Chemical Biology and Fundamental Medicine, Siberian Branch, Russian Academy of Sciences, Lavrentyev Avenue 8, Novosibirsk 630090, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9367-807X","authenticated-orcid":false,"given":"Anna A.","family":"Nushtaeva","sequence":"additional","affiliation":[{"name":"Institute of Chemical Biology and Fundamental Medicine, Siberian Branch, Russian Academy of Sciences, Lavrentyev Avenue 8, Novosibirsk 630090, Russia"},{"name":"Research Center for Genetics and Life Sciences, Sirius University of Science and Technology, Krasnodar Region, Sirius (Federal Territory of Sirius), Sochi 354340, Russia"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"methodology","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0009-0005-1477-1118","authenticated-orcid":false,"given":"Anastasia A.","family":"Leonteva","sequence":"additional","affiliation":[{"name":"Institute of Chemical Biology and Fundamental Medicine, Siberian Branch, Russian Academy of Sciences, Lavrentyev Avenue 8, Novosibirsk 630090, Russia"},{"name":"Research Center for Genetics and Life Sciences, Sirius University of Science and Technology, Krasnodar Region, Sirius (Federal Territory of Sirius), Sochi 354340, Russia"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"methodology","vocabulary":"credit"}]},{"given":"Yulya I.","family":"Savinovskaya","sequence":"additional","affiliation":[{"name":"Institute of Chemical Biology and Fundamental Medicine, Siberian Branch, Russian Academy of Sciences, Lavrentyev Avenue 8, Novosibirsk 630090, Russia"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"methodology","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2420-0483","authenticated-orcid":false,"given":"Galina V.","family":"Kochneva","sequence":"additional","affiliation":[{"name":"State Research Center of Virology and Biotechnology \u201cVector\u201d, Rospotrebnadzor, Koltsovo 630559, Russia"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"resources","vocabulary":"credit"},{"role":"data-curation","vocabulary":"credit"}]},{"given":"Vladimir A.","family":"Richter","sequence":"additional","affiliation":[{"name":"Institute of Chemical Biology and Fundamental Medicine, Siberian Branch, Russian Academy of Sciences, Lavrentyev Avenue 8, Novosibirsk 630090, Russia"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"project-administration","vocabulary":"credit"},{"role":"funding-acquisition","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3145-1878","authenticated-orcid":false,"given":"Elena V.","family":"Kuligina","sequence":"additional","affiliation":[{"name":"Institute of Chemical Biology and Fundamental Medicine, Siberian Branch, Russian Academy of Sciences, Lavrentyev Avenue 8, Novosibirsk 630090, Russia"},{"name":"Oncostar LLC, Inzhenernaya Street 23, Novosibirsk 630090, Russia"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"writing-review-editing","vocabulary":"credit"},{"role":"supervision","vocabulary":"credit"},{"role":"funding-acquisition","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8062-5689","authenticated-orcid":false,"given":"Dmitriy V.","family":"Semenov","sequence":"additional","affiliation":[{"name":"Institute of Chemical Biology and Fundamental Medicine, Siberian Branch, Russian Academy of Sciences, Lavrentyev Avenue 8, Novosibirsk 630090, Russia"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"conceptualization","vocabulary":"credit"},{"role":"methodology","vocabulary":"credit"},{"role":"software","vocabulary":"credit"},{"role":"formal-analysis","vocabulary":"credit"},{"role":"investigation","vocabulary":"credit"},{"role":"writing-original-draft","vocabulary":"credit"},{"role":"supervision","vocabulary":"credit"},{"role":"project-administration","vocabulary":"credit"}]}],"member":"1968","published-online":{"date-parts":[[2026,9,11]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"2563","DOI":"10.1200\/JCO.2005.04.5963","article-title":"Radiotherapy and Temozolomide for Newly Diagnosed Glioblastoma: Recursive Partitioning Analysis of the EORTC 26981\/22981-NCIC CE3 Phase III Randomized Trial","volume":"24","author":"Mirimanoff","year":"2006","journal-title":"J. Clin. Oncol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"67","DOI":"10.1007\/s11910-021-01153-8","article-title":"The Evolving Classification of Diffuse Gliomas: World Health Organization Updates for 2021","volume":"21","author":"Perez","year":"2021","journal-title":"Curr. Neurol. Neurosci. Rep."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"19","DOI":"10.1093\/ijpp\/riae054","article-title":"Immune Checkpoint Inhibitors in Cancer Patients from the Perspective of Pharmaceutical Care: A Scoping Review","volume":"33","author":"Lira","year":"2025","journal-title":"Int. J. Pharm. Pract."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"546","DOI":"10.1080\/21645515.2020.1782692","article-title":"PD-1\/PD-L1 Immune Checkpoint Inhibitors in Glioblastoma: Clinical Studies, Challenges and Potential","volume":"17","author":"Yang","year":"2021","journal-title":"Hum. Vaccines Immunother."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"vdab066","DOI":"10.1093\/noajnl\/vdab066","article-title":"Anti-PD-1 Checkpoint Blockade Monotherapy in the Orthotopic GL261 Glioma Model: The Devil Is in the Detail","volume":"3","author":"Tritz","year":"2021","journal-title":"Neuro-Oncol. Adv."},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Ageenko, A.B., Vasileva, N.S., Chesnokova, A.S., Semenov, D.V., Byvakina, A.A., Dymova, M.A., Sen\u2019kova, A.V., Nushtaeva, A.A., Leonteva, A.A., and Savinovskaya, Y.I. (2026). Oncolytic Virus VV-GMCSF-Lact and Human GM-CSF Against GL261 Glioma in Immunocompetent Mice. Pharmaceuticals, 19.","DOI":"10.3390\/ph19030434"},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Semenov, D., Vasileva, N., Menyailo, M., Mishinov, S., Savinovskaya, Y., Ageenko, A., Chesnokova, A., Dymova, M., Stepanov, G., and Kochneva, G. (2025). Single-Cell Transcriptomic Changes in Patient-Derived Glioma and U87 Glioblastoma Cell Cultures Infected with the Oncolytic Virus VV-GMCSF-Lact. Int. J. Mol. Sci., 26.","DOI":"10.3390\/ijms26146983"},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Ageenko, A., Vasileva, N., Richter, V., and Kuligina, E. (2024). Combination of Oncolytic Virotherapy with Different Antitumor Approaches against Glioblastoma. Int. J. Mol. Sci., 25.","DOI":"10.3390\/ijms25042042"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"15","DOI":"10.1093\/bioinformatics\/bts635","article-title":"STAR: Ultrafast Universal RNA-Seq Aligner","volume":"29","author":"Dobin","year":"2013","journal-title":"Bioinformatics"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"550","DOI":"10.1186\/s13059-014-0550-8","article-title":"Moderated Estimation of Fold Change and Dispersion for RNA-Seq Data with DESeq2","volume":"15","author":"Love","year":"2014","journal-title":"Genome Biol."},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Chen, E.Y., Tan, C.M., Kou, Y., Duan, Q., Wang, Z., Meirelles, G.V., Clark, N.R., and Ma\u2019ayan, A. (2013). Enrichr: Interactive and Collaborative HTML5 Gene List Enrichment Analysis Tool. BMC Bioinform., 14.","DOI":"10.1186\/1471-2105-14-128"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1083","DOI":"10.1093\/bioinformatics\/btt090","article-title":"DeconRNASeq: A Statistical Framework for Deconvolution of Heterogeneous Tissue Samples Based on mRNA-Seq Data","volume":"29","author":"Gong","year":"2013","journal-title":"Bioinformatics"},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Semenov, D.V., Vasileva, N.S., Dymova, M.A., Mishinov, S.V., Savinovskaya, Y.I., Ageenko, A.B., Dome, A.S., Zinchenko, N.D., Stepanov, G.A., and Kochneva, G.V. (2023). Transcriptome Changes in Glioma Cells upon Infection with the Oncolytic Virus VV-GMCSF-Lact. Cells, 12.","DOI":"10.3390\/cells12222616"},{"key":"ref_14","first-page":"112768","article-title":"Interferon-Gamma Signaling Pathway: Modulation of Key Genes in the Progression of Glioblastoma","volume":"16","year":"2025","journal-title":"World J. Biol. Chem."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1189","DOI":"10.1016\/j.celrep.2017.04.031","article-title":"Interferon Receptor Signaling Pathways Regulating PD-L1 and PD-L2 Expression","volume":"19","author":"Shin","year":"2017","journal-title":"Cell Rep."},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Takacs, G.P., Kreiger, C.J., Luo, D., Tian, G., Garcia, J.S., Deleyrolle, L.P., Mitchell, D.A., and Harrison, J.K. (2023). Glioma-Derived CCL2 and CCL7 Mediate Migration of Immune Suppressive CCR2+\/CX3CR1+ M-MDSCs into the Tumor Microenvironment in a Redundant Manner. Front. Immunol., 13.","DOI":"10.3389\/fimmu.2022.993444"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"11025","DOI":"10.1038\/s41598-025-96302-1","article-title":"Multi-Omic Analyses Reveal PTPN6\u2019s Impact on Tumor Immunity across Various Cancers","volume":"15","author":"Zhong","year":"2025","journal-title":"Sci. Rep."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"90","DOI":"10.20892\/j.issn.2095-3941.2016.0086","article-title":"Roles of Rap1 Signaling in Tumor Cell Migration and Invasion","volume":"14","author":"Zhang","year":"2017","journal-title":"Cancer Biol. Med."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"64","DOI":"10.1007\/s11033-023-09062-4","article-title":"Upregulation of the Canonical Signaling Pathway of Interferon-Gamma Is Associated with Glioblastoma Progression","volume":"51","year":"2024","journal-title":"Mol. Biol. Rep."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"352","DOI":"10.20517\/jtgg.2025.90","article-title":"Persistent IFN-\u03b3 Signaling in Acquired Resistance to PD-(L)1 Blockade in NSCLC","volume":"9","author":"Karri","year":"2025","journal-title":"J. Transl. Genet. Genom."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"156","DOI":"10.1038\/s41392-023-01407-6","article-title":"Oncolytic Virotherapy: Basic Principles, Recent Advances and Future Directions","volume":"8","author":"Lin","year":"2023","journal-title":"Signal Transduct. Target. Ther."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"e1249561","DOI":"10.1080\/2162402X.2016.1249561","article-title":"Compensatory Upregulation of PD-1, LAG-3, and CTLA-4 Limits the Efficacy of Single-Agent Checkpoint Blockade in Metastatic Ovarian Cancer","volume":"6","author":"Huang","year":"2017","journal-title":"OncoImmunology"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"101711","DOI":"10.1016\/j.gendis.2025.101711","article-title":"The Significance of Epithelial\u2013Mesenchymal Transition (EMT) in the Initiation, Plasticity, and Treatment of Glioblastoma","volume":"13","author":"Xia","year":"2026","journal-title":"Genes Dis."},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Colardo, M., Segatto, M., and Di Bartolomeo, S. (2021). Targeting RTK-PI3K-mTOR Axis in Gliomas: An Update. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms22094899"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1009","DOI":"10.1158\/1078-0432.CCR-22-1031","article-title":"Cancer-Associated Fibroblasts in Inflammation and Antitumor Immunity","volume":"29","author":"Kennel","year":"2023","journal-title":"Clin. Cancer Res."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"2695","DOI":"10.1007\/s00018-018-2826-8","article-title":"The Impact of Phosphatases on Proliferative and Survival Signaling in Cancer","volume":"75","author":"Narla","year":"2018","journal-title":"Cell. Mol. Life Sci."},{"key":"ref_27","first-page":"2333","article-title":"A Combination of Protein Phosphatase 2A Inhibition and Checkpoint Immunotherapy: A Perfect Storm","volume":"18","author":"Clark","year":"2024","journal-title":"Mol. Oncol."},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Teft, W.A., Chau, T.A., and Madrenas, J. (2009). Structure-Function Analysis of the CTLA-4 Interaction with PP2A. BMC Immunol., 10.","DOI":"10.1186\/1471-2172-10-23"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"346","DOI":"10.1080\/01652176.1986.9694067","article-title":"Laboratory Animal Protection: The European Convention and the Dutch Act","volume":"8","author":"Rozemond","year":"1986","journal-title":"Vet. Q."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/27\/18\/8078\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,11]],"date-time":"2026-09-11T08:20:48Z","timestamp":1789114848000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/27\/18\/8078"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,9,11]]},"references-count":29,"journal-issue":{"issue":"18","published-online":{"date-parts":[[2026,9]]}},"alternative-id":["ijms27188078"],"URL":"https:\/\/doi.org\/10.3390\/ijms27188078","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,9,11]]}}}