Optimizing CAR-T Therapy through CRISPR technology

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Abstract: Cancer continues to be the leading cause of death worldwide, driving an urgent need for effective and precise therapeutic strategies. The combination of CRISPR-Cas9 gene-editing technology with chimeric antigen receptor T-cell (CAR-T) therapy has represented a breakthrough in cancer treatment. This paper offers an in-depth overview of the application of CRISPR technology in enhancing CAR-T therapy. Through targeting and disrupting genes such as TRAC and B2M in T cells, universal CAR-T cells are developed, mitigating immune rejection issues in allogeneic treatments. Epigenetic editing is utilized to modulate proto-oncogene enhancers, inhibiting tumor growth. Moreover, controllable Cas9 systems, including the Rimiducid safety switch, are employed to finely tune CAR-T activity, reducing side effects like cytokine release syndrome. Studies demonstrate that CRISPR technology has significantly improved the precision, safety, and adaptability of CAR-T therapy. However, challenges remain, such as off-target effects, insufficient delivery efficiency, and limited efficacy in treating solid tumors. Looking ahead, it will be crucial to develop high-fidelity CRISPR tools like base editors, optimize targeted delivery methods, and expand clinical trials for solid tumors. As technology advances, CRISPR-enhanced CAR-T therapy is expected to become a key component of personalized cancer treatment, providing patients with better chances of recovery.
Keywords: CRISPR/Cas9; CAR-T; tumor therapy.
APA Citation: Yuchen Liu (2025). Optimizing CAR-T Therapy through CRISPR technology. Transactions on Materials, Biotechnology and Life Sciences, 8(1), 71-76. https://doi.org/10.62051/ma2f5v65

References

  1. Nishimasu H, Ran FA, Hsu PD, et al. Crystal structure of Cas9 in complex with guide RNA and target DNA. Cell, 2014, 156(5): 935-949.
  2. Ding Y, Li H, Chen LL, Xie K. Recent advances in genome editing using CRISPR/Cas9. Frontiers in Plant Science, 2016, 7: 703.
  3. Cradick TJ, Fine EJ, Antico CJ, Bao G. CRISPR/Cas9 systems targeting β-globin and CCR5 genes have substantial off-target activity. Nucleic Acids Research, 2013, 41(20): 9584-9592.
  4. Fu Y, Foden JA, Khayter C, et al. High-frequency off-target mutagenesis induced by CRISPR-Cas nucleases in human cells. Nature Biotechnology, 2013, 31(9): 822-826.
  5. Hsu PD, Scott DA, Weinstein JA, et al. DNA targeting specificity of RNA-guided Cas9 nucleases. Nature Biotechnology, 2013, 31(9): 827-832.
  6. Mali P, Aach J, Stranges PB, et al. CAS9 transcriptional activators for target specificity screening and paired nickases for cooperative genome engineering. Nature Biotechnology, 2013, 31(9): 833-838.
  7. Pattanayak V, Lin S, Guilinger JP, et al. High-throughput profiling of off-target DNA cleavage reveals RNA-programmed Cas9 nuclease specificity. Nature Biotechnology, 2013, 31(9): 839-843.
  8. Tsai S, Zheng Z, Nguyen N, et al. GUIDE-seq enables genome-wide profiling of off-target cleavage by CRISPR-Cas nucleases. Nature Biotechnology, 2015, 33: 187-197.
  9. Kuscu C, Arslan S, Singh R, Thorpe J, Adli M. Genome-wide analysis reveals characteristics of off-target sites bound by the Cas9 endonuclease. Nature Biotechnology, 2014, 32(7): 677-683.
  10. Maude SL, Laetsch TW, Buechner J, et al. Tisagenlecleucel in children and young adults with B-cell lymphoblastic leukemia. The New England Journal of Medicine, 2018, 378(5): 439-448.
  11. Neelapu SS, Locke FL, Bartlett NL, et al. Axicabtagene ciloleucel CAR T-cell therapy in refractory large B-cell lymphoma. The New England Journal of Medicine, 2017, 377(26): 2531-2544.
  12. Locke FL, Ghobadi A, Jacobson CA, et al. Long-term safety and activity of axicabtagene ciloleucel in refractory large B-cell lymphoma (ZUMA-1): a single-arm, multicentre, phase 1-2 trial. The Lancet Oncology, 2019, 20(1): 31-42.
  13. Sadelain M, Brentjens R, Riviere I. The basic principles of chimeric antigen receptor design. Cancer Discovery, 2013, 3(4): 388-398.
  14. June CH, Sadelain M. Chimeric antigen receptor therapy. The New England Journal of Medicine, 2018, 379(1): 64-73.
  15. Pegram HJ, Lee JC, Hayman EG, et al. Tumor-targeted T cells modified to secrete IL-12 eradicate systemic tumors without need for prior conditioning. Blood, 2012, 119(18): 4133-4141.
  16. Qasim W, Zhan H, Samarasinghe S, et al. Molecular remission of infant B-ALL after infusion of universal TALEN gene-edited CAR T cells. Science Translational Medicine, 2017, 9(374): eaaj2013.
  17. Newick K, O'Brien S, Moon E, Albelda SM. CAR T cell therapy for solid tumors. Annual Review of Medicine, 2017, 68: 139-152.
  18. Adusumilli PS, Cherkassky L, Villena-Vargas J, et al. regional delivery of mesothelin-targeted CAR T cell therapy generates potent and long-lasting CD4-dependent tumor immunity. Science Translational Medicine, 2014, 6(261): 261ra151.
  19. Rafiq S, Yeku OO, Jackson HJ, et al. Targeted delivery of a PD-1-blocking scFv by CAR-T cells enhances anti-tumor efficacy in vivo. Nature Biotechnology, 2018, 36(9): 847-856.
  20. Stadtmauer EA, Fraietta JA, Davis MM, et al. CRISPR-engineered T cells in patients with refractory cancer. Science, 2020, 367(6481): eaba7365.
  21. Ren J, Liu X, Fang C, et al. Multiplex genome editing to generate universal CAR T cells resistant to PD1 inhibition. Clinical Cancer Research, 2017, 23(9): 2255-2266.
  22. Liu X, Zhang Y, Cheng C, et al. CRISPR-Cas9-mediated multiplex gene editing in CAR-T cells. Cell Research, 2017, 27(1): 154-157.
  23. Li K, Liu Y, Cao H, et al. Interrogation of enhancer function by enhancer-targeting CRISPR epigenetic editing. Nature Communications, 2020, 11(1): 485.
  24. Zheng Y, Nandakumar KS, Cheng K. Optimization of CAR-T cell-based therapies using small-molecule-based safety switches. Journal of Medicinal Chemistry, 2021, 64(14): 9577-9591.