Gene Therapy in Glioblastoma: From Vector Engineering to Immune Microenvironment Reprogramming

Author's Information:

Alper DEMIREZEN

Kocaeli Üniversitesi Tıbbi Biyoloji Anabilim Dalı

Sumeyye Seher Karaman

Tübitak Gebze Yerleşkesi Arge ve İnovasyon Merkezi

Vol 03 No 07 (2026):Volume 03 Issue 07 July 2026

Page No.: 340-351

Crossref DOI: https://doi.org/10.55677/IJMSPR/2026-3050-I701 | Published: 07 July, 2026 | | Google Scholar | Google

Abstract:

Glioblastoma (GBM) remains the most aggressive primary malignant brain tumor in adults, characterized by extensive intratumoral heterogeneity, diffuse infiltration, profound immunosuppression, and an invariably poor prognosis despite multimodal treatment strategies. Although maximal safe surgical resection followed by radiotherapy and temozolomide constitutes the current standard of care, therapeutic resistance and tumor recurrence remain nearly universal, underscoring the urgent need for innovative treatment modalities. In this context, gene therapy has emerged as a promising therapeutic paradigm capable of addressing several biological limitations associated with conventional therapies through targeted genetic modulation and immune microenvironment reprogramming. This narrative review comprehensively summarizes the advances achieved between 2016 and 2026 in GBM-directed gene therapy. Particular emphasis is placed on the engineering principles, biological characteristics, and translational applications of viral vectors including retroviral, adenoviral, and adeno-associated viral systems as well as emerging non-viral delivery platforms such as lipid nanoparticles, polymeric nanocarriers, electroporation, engineered exosomes, messenger RNA technologies, and CRISPR/Cas-based genome editing. Furthermore, the review discusses recent developments in oncolytic virotherapy, suicide gene systems, RNA interference strategies, immune-stimulatory gene delivery, and adoptive cellular gene therapies. Beyond vector technology, this review highlights the critical role of immune microenvironment remodeling in improving therapeutic efficacy. Current evidence suggests that successful gene therapy should not solely focus on tumor cell eradication but should also promote durable antitumor immunity through enhanced antigen presentation, increased lymphocyte infiltration, reversal of myeloid-mediated immunosuppression, and rational integration with immune checkpoint blockade, dendritic cell vaccines, CAR-T cell therapy, and other multimodal immunotherapeutic approaches. Finally, ongoing clinical trials, translational challenges, manufacturing considerations, biomarker-guided patient selection, and future perspectives are critically discussed to provide a comprehensive overview of the evolving landscape of gene therapy for glioblastoma.

KeyWords:

Glioblastoma; Gene therapy; Viral vectors; Oncolytic virotherapy; CAR-T cells; CRISPR/Cas9; RNA therapeutics; Immune microenvironment; Translational medicine.

References:

  1. Akgeyik F, Demirezen A, Erbaş O. Glioblastoma multiforme: Subtypes based on IDH, ATRX, H3F3A, MGMT and p53 mutations. D J Med Sci 2024;10(1):34-41. doi: 10.5606/fng.btd.2024.144.
  2. Rasheed Z, Ma YK, Ullah I, Al Shloul T, Tufail AB, Ghadi YY, et al. Automated classification of brain tumors from magnetic resonance ımaging using deep learning. Brain Sci 2023;13:602. doi: 10.3390/ brainsci13040602.
  3. S Tandel G, Biswas M, G Kakde O, Tiwari A, S Suri H, Turk M, et al. A review on a deep learning perspective in brain cancer classification. Cancers (Basel) 2019;11:111. doi: 10.3390/ cancers11010111.
  4. Nadeem MW, Ghamdi MAA, Hussain M, Khan MA, Khan KM, Almotiri SH, et al. Brain tumor analysis empowered with deep learning: A review, taxonomy, and future challenges. Brain Sci 2020;10:118. doi: 10.3390/brainsci10020118.
  5. Li W, Zhang S, Xing D, Qin H. Pulsed microwave ınduced thermoacoustic shockwave for precise glioblastoma therapy with the skin and skull intact. Small 2022;18:e2201342. doi: 10.1002/ smll.202201342.
  6. Louis DN, Perry A, Reifenberger G, von Deimling A, Figarella-Branger D, Cavenee WK, et al. The 2016 World Health Organization classification of tumors of the central nervous system: A summary. Acta Neuropathol 2016;131:803-20. doi: 10.1007/s00401 016-1545-1.
  7. Piroth MD, Galldiks N, Pinkawa M, Holy R, Stoffels G, Ermert J, et al. Relapse patterns after radiochemotherapy of glioblastoma with FET PET guided boost irradiation and simulation to optimize radiation target volume. Radiat Oncol 2016;11:87. doi: 10.1186/s13014-016-0665-z.
  8. Weller M, van den Bent M, Preusser M, Le Rhun E, Tonn JC, Minniti G, et al. EANO guidelines on the diagnosis and treatment of diffuse gliomas of adulthood. Nat Rev Clin Oncol. 2021;18:170-86.
  9. Sahm F, Capper D, Jeibmann A, Hovestadt V, von Deimling A, Weller M, et al. Molecular diagnostic tools for the World Health Organization 2021 classification of gliomas. Neuro Oncol. 2023;25:1731-49.
  10. Stupp R, Taillibert S, Kanner A, Read W, Steinberg DM, Lhermitte B, et al. Effect of tumor-treating fields plus maintenance temozolomide vs maintenance temozolomide alone on survival in patients with glioblastoma: a randomized clinical trial. JAMA. 2017;318:2306-16. 
  11. Reardon DA, Brandes AA, Omuro A, Mulholland P, Lim M, Wick A, et al. Effect of nivolumab vs bevacizumab in patients with recurrent glioblastoma: the CheckMate 143 phase 3 randomized clinical trial. JAMA Oncol. 2020;6:1003-10.
  12. Choi BD, Gerstner ER, Frigault MJ, Leick MB, Mount CW, Balaj L, et al. Intraventricular CARv3-TEAM-E T cells in recurrent glioblastoma. N Engl J Med. 2024;390:1290-8.
  13. Uchida M, Shimatsu Y, Onoe K, Matsuyama N, Niki R, Ikeda JE, et al. Production of transgenic miniature pigs by pronuclear microinjection Transgenic Res, 2001; 10 (6): 577-82. Doi: 10.1023/A:1013059917280.
  14. Hofmann A, Zakhartchenko V, Weppert M, Sebald H, Wenigerkind H, Brem G, et al. Generation of transgenic cattle by lentiviral gene transfer into oocytes. Biol Reprod, 2004; 71 (2): 405-9. Doi: 10.1095/biolreprod.104.028472.
  15. Ibraheem D, Elaissari A, Fessi H. Gene therapy and DNA delivery systems. Int J Pharm, 2014; 459: 70-83. Doi: 10.1016/j.ijpharm.2013.11.041.
  16. Gehl J. Electroporation: Theory and methods, perspectives for drug delivery, gene therapy and research. Acta Physiol Scand, 2003; 177 (4): 437- Doi: 10.1046/j.1365-201X.2003.01093.x.
  17. Naldini L. Gene therapy returns to centre stage.Nature. 2015;526(7573):351–360. doi:10.1038/nature15818.
  18. Milone MC, O'Doherty U. Clinical use of lentiviral vectors.Leukemia. 2018;32:1529–1541. doi:10.1038/s41375-018-0106-0.
  19. Dunbar CE, High KA, Joung JK, Kohn DB, Ozawa K, Sadelain M. Gene therapy comes of age. Science. 2018;359:eaan4672. doi:10.1126/science.aan4672.
  20. Hacein-Bey-Abina S, et al. LMO2-associated clonal T-cell proliferation in two patients after gene therapy for SCID-X1. Science. 2003;302:415–419. doi:10.1126/science.1088547.
  21. Wold WSM, Toth K. Adenovirus vectors for gene therapy, vaccination and cancer gene therapy. Curr Gene Ther. 2013;13:421–433.
  22. Ginn SL, Amaya AK, Alexander IE, Edelstein ML, Abedi MR. Gene therapy clinical trials worldwide to 2017. J Gene Med. 2018;20:e3015.
  23. Crystal RG. Adenovirus: the first effective in vivo gene delivery vector. Hum Gene Ther. 2014;25:3–11.
  24. Lee CS, Bishop ES, Zhang R, et al. Adenovirus-mediated gene delivery. Genes Dis. 2017;4:43–63.
  25. Naso MF, Tomkowicz B, Perry WL III, Strohl WR. Adeno-associated virus (AAV) as a vector for gene therapy. BioDrugs. 2017;31:317–334.
  26. Wang D, Tai PWL, Gao G. Adeno-associated virus vector as a platform for gene therapy delivery. Nat Rev Drug Discov. 2019;18:358–378.
  27. Keeler AM, Flotte TR. Recombinant adeno-associated virus gene therapy. Gene Ther. 2019;26:473–481.
  28. Wang L, et al. Clinical development of AAV gene therapy. Signal Transduct Target Ther. 2022;7:150.
  29. Wang D, Gao G. State-of-the-art AAV gene therapy.Nat Rev Drug Discov. 2024.
  30. Bulcha JT, Wang Y, Ma H, Tai PWL, Gao G. Viral vector platforms within the gene therapy landscape. Signal Transduct Target Ther. 2021;6:53.
  31. Mitragotri S, et al. Drug delivery research for the future.Nat Rev Drug Discov. 2021;20:101–124.
  32. Kulkarni JA, et al. Lipid nanoparticles enabling gene therapies. Nat Nanotechnol. 2021;16:630–643.
  33. Hou X, Zaks T, Langer R, Dong Y. Lipid nanoparticles for mRNA delivery. Nat Rev Mater. 2021;6:1078–1094.
  34. Ramamoorth M, Narvekar A. Non viral vectors in gene therapy—An overview. J Clin Diagn Res. 2015;9:GE01–GE06.
  35. Desjardins A, Gromeier M, Herndon JE 2nd, Beaubier N, Bolognesi DP, Friedman AH, et al. Recurrent glioblastoma treated with recombinant poliovirus. N Engl J Med. 2018;379:150-61.
  36. Todo T, Ito H, Ino Y, Ohtsu H, Ota Y, Shibahara J, et al. Intratumoral oncolytic herpes virus G47Delta for residual or recurrent glioblastoma: a phase 2 trial. Nat Med. 2022;28:1630-9.
  37. Frampton JE. Teserpaturev/G47Delta: first approval. BioDrugs. 2022;36:667-72.
  38. Ling AL, Solomon IH, Montalvo Landivar A, Frosch MP, Chiocca EA, Reardon DA, et al. Clinical trial links oncolytic immunoactivation to survival in glioblastoma. Nature. 2023;623:157-66.
  39. Nassiri F, Patil V, Yefet LS, Singh O, Liu J, Dang RMA, et al. Oncolytic DNX-2401 virotherapy plus pembrolizumab in recurrent glioblastoma: a phase 1/2 trial. Nat Med. 2023;29:1370-8.
  40. Cloughesy TF, Petrecca K, Walbert T, Butowski N, Salacz M, Perry J, et al. Effect of vocimagene amiretrorepvec in combination with flucytosine vs standard of care on survival following tumor resection in patients with recurrent high-grade glioma: a randomized clinical trial. JAMA Oncol. 2020;6:1939-46.
  41. Umemura Y, Orringer D, Junck L, Chenevert T, Mikkelsen T, Lowenstein PR, et al. Combined cytotoxic and immune-stimulatory gene therapy for primary adult high-grade glioma: a phase 1, first-in-human trial. Lancet Oncol. 2023;24:1042-52.
  42. He W, Tan Y, Yang X, Zhao X, Li C, Yu H, et al. Engineering nanomedicine for non-viral RNA-based glioblastoma therapy. Pharmaceutics. 2024;16:482.
  43. Sharma RK, Calderon C, Kaur B, Vrionis F, Batra SK, Raizer JJ, et al. Targeting non-coding RNA for glioblastoma therapy. Front Med Technol. 2021;3:678593.
  44. Li S, Meng C, Ruan H, Liu X, Luo R, Huang T, et al. Brain-targeted biomimetic siRNA nanoparticles for drug resistance reversal in glioblastoma. J Control Release. 2024;370:396-410.
  45. Liang SF, Zhu Y, Sun Y, Huang X, Wang J, Xu H, et al. Delivery of siRNA based on engineered exosomes for glioblastoma therapy by targeting STAT3. Biomater Sci. 2022;10:1581-93.
  46. Begagic E, Avdic D, Ramaswamy V, Mekic-Abazovic A, Abazovic A, Rotim K, et al. CRISPR/Cas9-mediated gene therapy for glioblastoma: a scoping review. Int J Mol Sci. 2024;25:960.
  47. Fang Y, Wang D, Wang J, Li J, Wang Q, Cheng Z. Unlocking glioblastoma vulnerabilities with CRISPR-based functional genomics. Int J Mol Sci. 2024;25:5702.
  48. Liu Y, Wang H, Chen Q, Zhang J, Wang Y, Wang H, et al. Immunotherapy for glioblastoma: current state, challenges and future perspectives. Cell Mol Immunol. 2024;21:1209-35.
  49. Lin H, Patel S, Affleck K, Tsai J, Yang H, Yao Y. Understanding the immunosuppressive microenvironment of glioblastoma. J Hematol Oncol. 2024;17:41.
  50. Nader NE, Venkatesh HS, Johnson MO. Barriers to T cell functionality in the glioblastoma microenvironment. Cancers (Basel). 2024;16:3273.
  51. Sarantopoulos A, Sonabend AM, Lesniak MS, Horbinski C. Therapeutic approaches to modulate the immune microenvironment in gliomas. NPJ Precis Oncol. 2024;8:245.
  52. Chiocca EA, Yu JS, Lukas RV, Solomon IH, Ligon KL, Nakashima H, et al. Regulatable interleukin-12 gene therapy in patients with recurrent high-grade glioma: results of a phase 1 trial. Sci Transl Med. 2019;11:eaaw5680.
  53. Chiocca EA, Gelb AB, Chen CC, Rao G, Reardon DA, Wen PY, et al. Combined immunotherapy with controlled interleukin-12 gene therapy and immune checkpoint blockade in recurrent glioblastoma. Neuro Oncol. 2021;23:951-62.
  54. von Roemeling CA, Wang Y, Qie Y, Chen D, Miska J, Juarez J, et al. Adeno-associated virus delivered CXCL9 sensitizes glioblastoma to anti-PD-1 immune checkpoint blockade. Nat Commun. 2024;15:5661.
  55. Chongsathidkiet P, Jackson C, Koyama S, Loebel F, Cui X, Farber SH, et al. Sequestration of T cells in bone marrow in the setting of glioblastoma and other intracranial tumors. Nat Med. 2018;24:1459-68.
  56. Friedman GK, Johnston JM, Bag AK, Bernstock JD, Li R, Aban I, et al. Oncolytic HSV-1 G207 immunovirotherapy for pediatric high-grade gliomas. N Engl J Med. 2021;384:1613-22.
  57. Hossain JA, Marchini A, Fehse B, Bjerkvig R, Miletic H. Suicide gene therapy for the treatment of high-grade glioma: past lessons, present trends, and future prospects. Neurooncol Adv. 2020;2:vdaa013.
  58. Shah S, Ali A, Hsu F, Vora P, Siu IM, Aghi MK. Gene therapy for glioblastoma multiforme. Viruses. 2025;17:118.
  59. Jiang S, Zheng C, Pan Y, Yuan X, Yang Y, Xia Y. Clinical advances in oncolytic virus therapy for malignant glioma. Cancer Biol Med. 2023;20:857-73.
  60. Banerjee K, Núñez FJ, Haase S, McClellan BL, Faisal SM, Carney SV, Yu J, Alghamri MS, Asad AS, Candia AJN, Varela ML, Candolfi M, Lowenstein PR and Castro MG (2021) Current Approaches for Glioma Gene Therapy and Virotherapy. Front. Mol. Neurosci. 14:621831. doi: 10.3389/fnmol.2021.621831
  61. Li J, Wang W, Wang J, Cao Y, Wang S and Zhao J (2021) Viral Gene Therapy for Glioblastoma Multiforme: A Promising Hope for the Current Dilemma. Front. Oncol. 11:678226. doi: 10.3389/fonc.2021.678226
  62. Rayati, M., Mansouri, V., & Ahmadbeigi, N. (2024). Gene therapy in glioblastoma multiforme: Can it be a role changer? Heliyon, 10(5), e27087. https://doi.org/10.1016/j.heliyon.2024.e27087
  63. Xu, X., Chen, W., Zhu, W. et al. Adeno‐associated virus (AAV)-based gene therapy for glioblastoma. Cancer Cell Int 21, 76 (2021).  https://doi.org/10.1186/s12935-021-01776-4
  64. Caffery, B., Lee, J. S., & Alexander-Bryant, A. A. (2019). Vectors for Glioblastoma Gene Therapy: Viral & Non-Viral Delivery Strategies. Nanomaterials, 9(1), 105. https://doi.org/10.3390/nano9010105
  65. Rouatbi, N., Walters, A. A., Costa, P. M., Qin, Y., Liam-Or, R., Grant, V., Pollard, S. M., Wang, J. T., & Al-Jamal, K. T. (2024). RNA lipid nanoparticles as efficient in vivo CRISPR-Cas9 gene editing tool for therapeutic target validation in glioblastoma cancer stem cells. Journal of Controlled Release, 375, 776–787. https://doi.org/10.1016/j.jconrel.2024.09.019
  66. Yang, Q., Zhou, Y., Chen, J., Huang, N., Wang, Z., & Cheng, Y. (2021). Gene Therapy for Drug-Resistant Glioblastoma via Lipid-Polymer Hybrid Nanoparticles Combined with Focused Ultrasound. International Journal of Nanomedicine, 16, 185–199. https://doi.org/10.2147/IJN.S286221
Download PDF
Article Metrics

Views

5

Page No.

340-351

Published

07 July, 2026

Copyrights & License

License This work is licensed under a Creative Commons Attribution 4.0 International License.

Alper DEMIREZEN, Sumeyye Seher Karaman ©2026

All Content should be original and unpublished.

Most Read Articles

Relationship Issues, Emotion Regulation and Everyday Discrimination Among Transgender Individuals

Vol 02 No 06 (2025) 667 views

Spectrophotometric Methods for Paracetamol Determination in Pharmaceutical Formulations: A Concise Analytical Review

Vol 2 No 7 (2025) 573 views

Trends and Determinants of Mortality Among Children in Nigeria: A Teaching Hospital Perspective

Vol 02 No 08 (2025) 534 views

Assessment of Over-the-Counter Drug Use Among Pregnant Women in Damaturu, Northeast Nigeria

Vol 2 No 7 (2025) 533 views

Perceived Patterns of Vaccine-Preventable Diseases and Barriers to Immunization Uptake in Isoko North and South LGAs Of Delta State

Vol 2 No 7 (2025) 516 views