CLDN6-CAR-T Cell Therapy in Ovarian Cancer: Overcoming the Challenges of Tumour Heterogeneity
DOI:
https://doi.org/10.54097/q8ry4c85Keywords:
CLDN6-CAR-T, Immunotherapy, Ovarian Cancer, HeterogeneityAbstract
Chimeric antigen receptor T cells (CAR-T cells) are synthetically modified versions of T lymphocytes with an additional CAR that allows specific, customised antigen binding and subsequent cell elimination by apoptosis. For ovarian cancer, Claudin-6 (CLDN6) is an excellent target antigen for CAR-T therapy, due to its high expression within ovarian cancer tissue and low expression in healthy adult cells. Scientists have therefore developed CLDN6-specific CAR-T cell therapy (CLDN6-CAR-T). CLDN6-CAR-T offers benefits compared to traditional surgery and chemotherapy, but faces immature clinical outcomes and heterogeneity-related limitations, including antigen escape, TME immunosuppression, and cancer-associated fibroblasts. In the face of these challenges, possible solutions currently being examined and developed include multi-target therapy, CXCR2-targeting, bionic physical barriers, and the use of heparanase.
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[1] A. C. Veneziani et al., “Heterogeneity and treatment landscape of ovarian carcinoma,” Nature Reviews Clinical Oncology, vol. 20, no. 12, pp. 820–842, Dec. 2023, doi: https://doi.org/10.1038/s41571-023-00819-1.
[2] Ovarian Cancer Action, “Types of ovarian cancer | Ovarian cancer information,” Ovarian Cancer Action, Jun. 03, 2024. https://ovarian.org.uk/ovarian-cancer/types-of-ovarian-cancer/
[3] R. C. Sterner and R. M. Sterner, “CAR-T Cell therapy: Current Limitations and Potential Strategies,” Blood Cancer Journal, vol. 11, no. 4, Apr. 2021, doi: https://doi.org/10.1038/s41408-021-00459-7.
[4] R. C. Larson and M. V. Maus, “Recent Advances and Discoveries in the Mechanisms and Functions of CAR T Cells,” Nature Reviews Cancer, vol. 21, no. 3, pp. 145–161, Jan. 2021, doi: https:// doi. org/ 10.1038/s41568-020-00323-z.
[5] J. Li et al., “NK-92MI Cells Engineered with Anti-claudin-6 Chimeric Antigen Receptors in Immunotherapy for Ovarian Cancer,” International Journal of Biological Sciences, vol. 20, no. 5, pp. 1578–1601, Feb. 2024, doi: https://doi.org/10.7150/ijbs.88539.
[6] National Cancer Institute, “CAR T Cells: Engineering Immune Cells to Treat Cancer,” Cancer.gov, Feb. 26, 2025. https://www.cancer.gov/about-cancer/treatment/research/car-t-cells#closing-in-on-car-t-cell-therapy-for-solid-tumors
[7] S. P. Blagden, “Harnessing Pandemonium: The Clinical Implications of Tumor Heterogeneity in Ovarian Cancer,” Frontiers in Oncology, vol. 5, Jun. 2015, doi: https://doi.org/10.3389/fonc.2015.00149.
[8] National Cancer Institute, “Surgery,” Cancer.gov, Apr. 29, 2015. https://www.cancer.gov/about-cancer/ treatment/ types/surgery
[9] A. Zafar, S. Khatoon, M. J. Khan, J. Abu, and A. Naeem, “Advancements and limitations in traditional anti-cancer therapies: a comprehensive review of surgery, chemotherapy, radiation therapy, and hormonal therapy,” Discover Oncology, vol. 16, no. 1, Apr. 2025, doi: https://doi.org/10.1007/s12672-025-02198-8.
[10] M. Chidambaram, R. Manavalan, and K. Kathiresan, “Nanotherapeutics to Overcome Conventional Cancer Chemotherapy Limitations,” Journal of Pharmacy & Pharmaceutical Sciences, vol. 14, no. 1, pp. 67–77, Feb. 2011, doi: https://doi.org/10.18433/J30C7D.
[11] H. E. Hughes-Parry, R. S. Cross, and M. R. Jenkins, “The Evolving Protein Engineering in the Design of Chimeric Antigen Receptor T Cells,” International Journal of Molecular Sciences, vol. 21, no. 1, p. 204, Dec. 2019, doi: https://doi.org/10.3390/ijms21010204.
[12] A. Alnefaie et al., “Chimeric Antigen Receptor T-Cells: An Overview of Concepts, Applications, Limitations, and Proposed Solutions,” Frontiers in Bioengineering and Biotechnology, vol. 10, p. 797440, Jun. 2022, doi: https://doi.org/10.3389/fbioe.2022.797440.
[13] H. Balke-Want et al., “Non-viral chimeric antigen receptor (CAR) T cells going viral,” Immuno-Oncology and Technology, vol. 18, p. 100375, Jun. 2023, doi: https://doi.org/10.1016/j.iotech.2023.100375.
[14] T. A. Bui, H. Mei, R. Sang, D. G. Ortega, and W. Deng, “Advancements and challenges in developing in vivo CAR T cell therapies for cancer treatment,” EBioMedicine, vol. 106, p. 105266, Aug. 2024, doi: https://doi.org/10.1016/j.ebiom.2024.105266.
[15] U. Ahmad et al., “Chimeric antigen receptor T cell structure, its manufacturing, and related toxicities; A comprehensive review,” Advances in Cancer Biology - Metastasis, vol. 4, p. 100035, Jul. 2022, doi: https://doi.org/10.1016/j.adcanc.2022.100035.
[16] Z. Zhao, Y. Chen, N. M. Francisco, Y. Zhang, and M. Wu, “The application of CAR-T cell therapy in hematological malignancies: advantages and challenges,” Acta Pharmaceutica Sinica B, vol. 8, no. 4, pp. 539–551, Jul. 2018, doi: https://doi.org/10.1016/j.apsb.2018.03.001.
[17] Myeloma UK, “Ask the Nurse: CAR-T cell treatments,” Myeloma UK, Feb. 2022. https://www. myeloma. org.uk/library/ask-the-nurse-car-t-cells/
[18] R. C. De Marco, H. J. Monzo, and P. M. Ojala, “CAR T Cell Therapy: A Versatile Living Drug,” International Journal of Molecular Sciences, vol. 24, no. 7, Mar. 2023, doi: https://doi.org/10.3390/ ijms 24076300.
[19] D. Günzel and A. S. L. Yu, “Claudins and the modulation of tight junction permeability,” Physiological reviews, vol. 93, no. 2, pp. 525–569, 2013, doi: https://doi.org/10.1152/physrev.00019.2012.
[20] G. Krause, L. Winkler, S. L. Mueller, R. F. Haseloff, J. Piontek, and I. E. Blasig, “Structure and function of claudins,” Biochimica et biophysica acta, vol. 1778, no. 3, pp. 631–645, 2007, doi: https://doi.org/ 10. 1016/ j.bbamem.2007.10.018.
[21] A. Berselli, F. Benfenati, L. Maragliano, and G. Alberini, “Multiscale modelling of claudin-based assemblies: A magnifying glass for novel structures of biological interfaces,” Computational and Structural Biotechnology Journal, vol. 20, pp. 5984–6010, Oct. 2022, doi: https://doi.org/ 10.1016/j. csbj. 2022.10.038.
[22] M. S. J. McDermott et al., “Preclinical Efficacy of the Antibody–Drug Conjugate CLDN6–23-ADC for the Treatment of CLDN6-Positive Solid Tumors,” Clinical Cancer Research, vol. 29, no. 11, pp. 2131–2143, 2023, doi: https://doi.org/10.1158/1078-0432.ccr-22-2981.
[23] H. Qu, J. Qiu, and C. Quan, “CLDN6: From Traditional Barrier Function to Emerging Roles in Cancers,” International Journal of Molecular Sciences, vol. 22, no. 24, Dec. 2021, doi: https://doi.org/10. 3390/ijms 222413416.
[24] L. Wang et al., “Clinicopathologic significance of claudin-6, occludin, and matrix metalloproteinases-2 expression in ovarian carcinoma,” Diagnostic Pathology, vol. 8, no. 1, Nov. 2013, doi: https://doi.org/ 10.1186/ 1746-1596-8-190.
[25] H. Du, X. Yang, J. Fan, and X. Du, “Claudin 6: Therapeutic prospects for tumours, and mechanisms of expression and regulation (Review),” Molecular Medicine Reports, vol. 24, no. 3, Jul. 2021, doi: https://doi.org/10.3892/mmr.2021.12316.
[26] L. Seidmann et al., “The Chimeric Antigen Receptor T Cell Target Claudin 6 Is a Marker for Early Organ-Specific Epithelial Progenitors and Is Expressed in Some Pediatric Solid Tumor Entities,” Cancers, vol. 17, no. 6, p. 920, Mar. 2025, doi: https://doi.org/10.3390/cancers17060920.
[27] A. Mackensen et al., “CLDN6-specific CAR-T cells plus amplifying RNA vaccine in relapsed or refractory solid tumors: the phase 1 BNT211-01 trial,” Nature Medicine, vol. 29, no. 11, pp. 2844–2853, Oct. 2023, doi: https://doi.org/10.1038/s41591-023-02612-0.
[28] F. Rieux-Laucat et al., “Inherited and Somatic CD3ζ Mutations in a Patient with T-Cell Deficiency,” The New England Journal of Medicine, vol. 354, no. 18, pp. 1913–1921, May 2006, doi: https://doi.org/ 10. 1056/nejmoa053750.
[29] R. Singh, Y.-H. Kim, S.-J. Lee, H.-S. Eom, and B. K. Choi, “4-1BB immunotherapy: advances and hurdles,” Experimental & Molecular Medicine, vol. 56, no. 1, pp. 32–39, Jan. 2024, doi: https://doi.org/ 10.1038/s12276-023-01136-4.
[30] C. Cutri-French, D. Nasioudis, E. George, and J. L. Tanyi, “CAR-T Cell Therapy in Ovarian Cancer: Where Are We Now?,” Diagnostics, vol. 14, no. 8, p. 819, Apr. 2024, doi: https://doi.org/10.3390/ diagnostics14080819.
[31] B. Screnci et al., “Antibody specificity against highly conserved membrane protein Claudin 6 driven by single atomic contact point,” iScience, vol. 25, no. 12, pp. 105665–105665, Nov. 2022, doi: https://doi. org/ 10.1016/j.isci.2022.105665.
[32] D. D. Bowtell et al., “Rethinking ovarian cancer II: reducing mortality from high-grade serous ovarian cancer,” Nature Reviews Cancer, vol. 15, no. 11, pp. 668–679, Oct. 2015, doi: https://doi.org/10.1038/ nrc4019.
[33] C. E. Meacham and S. J. Morrison, “Tumour heterogeneity and cancer cell plasticity,” Nature, vol. 501, no. 7467, pp. 328–337, Sep. 2013, doi: https://doi.org/10.1038/nature12624.
[34] M. Gerlinger et al., “Intratumor Heterogeneity and Branched Evolution Revealed by Multiregion Sequencing,” New England Journal of Medicine, vol. 366, no. 10, pp. 883–892, Mar. 2012, doi: https://doi.org/10.1056/nejmoa1113205.
[35] M. Greaves and C. C. Maley, “Clonal evolution in cancer,” Nature, vol. 481, no. 7381, pp. 306–313, Jan. 2012, doi: https://doi.org/10.1038/nature10762.
[36] D. Gumber and L. D. Wang, “Improving CAR-T immunotherapy: Overcoming the challenges of T cell exhaustion,” eBioMedicine, vol. 77, p. 103941, Mar. 2022, doi: https://doi.org/10. 1016/j.ebiom. 2022. 103941.
[37] M. Ijaz et al., “Overcoming barriers in glioblastoma: The potential of CAR T cell immunotherapy,” Theranostics, vol. 15, no. 14, pp. 7090–7126, Jun. 2025, doi: https://doi.org/10.7150/thno.114257.
[38] K. P. Lim et al., “CD4+CD25hiCD127low Regulatory T Cells Are Increased in Oral Squamous Cell Carcinoma Patients,” PLoS ONE, vol. 9, no. 8, p. e103975, Aug. 2014, doi: https://doi.org/10.1371/ journal. pone.0103975.
[39] Y. Qi, L. Zhang, Y. Liu, Y. Li, Y. Liu, and Z. Zhang, “Targeted modulation of myeloid-derived suppressor cells in the tumor microenvironment: Implications for cancer therapy,” Biomedicine & Pharmacotherapy, vol. 180, p. 117590, 2024, doi: https://doi.org/10.1016/j.biopha.2024.117590.
[40] M. Motallebnezhad, F. Jadidi-Niaragh, E. S. Qamsari, S. Bagheri, T. Gharibi, and M. Yousefi, “The immunobiology of myeloid-derived suppressor cells in cancer,” Tumor biology, vol. 37, no. 2, pp. 1387–1406, Nov. 2015, doi: https://doi.org/10.1007/s13277-015-4477-9.
[41] K. Newick, S. O’Brien, E. Moon, and S. M. Albelda, “CAR T Cell Therapy for Solid Tumors,” Annual Review of Medicine, vol. 68, no. 1, pp. 139–152, Jan. 2017, doi: https://doi.org/10.1146/annurev-med-062315-120245.
[42] M. Schwab, Encyclopedia of Cancer, 4th ed. Springer Berlin, Heidelberg, 2017. doi: https://doi.org/ 10.1007/ 978-3-662-46875-3.
[43] J. M. Pitt, A. Marabelle, A. Eggermont, J.-C. . Soria, G. Kroemer, and L. Zitvogel, “Targeting the tumor microenvironment: removing obstruction to anticancer immune responses and immunotherapy,” Annals of Oncology, vol. 27, no. 8, pp. 1482–1492, 2016, doi: https://doi.org/10.1093/annonc/mdw168.
[44] R. A. van Amerongen et al., “PRAME and CTCFL-reactive TCRs for the treatment of ovarian cancer,” Frontiers in immunology, vol. 14, Mar. 2023, doi: https://doi.org/10.3389/fimmu.2023.1121973.
[45] T. K. Bera and I. Pastan, “Mesothelin Is Not Required for Normal Mouse Development or Reproduction,” Molecular and Cellular Biology, vol. 20, no. 8, pp. 2902–2906, Apr. 2000, doi: https://doi.org/ 10.1128/ mcb.20.8.2902-2906.2000.
[46] M. Schwab, Encyclopedia of Cancer, 3rd ed. Springer Berlin, Heidelberg, 2011. doi: https://doi. org/10. 1007/978-3-642-16483-5.
[47] A. Morello, M. Sadelain, and P. S. Adusumilli, “Mesothelin-Targeted CARs: Driving T Cells to Solid Tumors,” Cancer Discovery, vol. 6, no. 2, pp. 133–146, Oct. 2015, doi: https://doi.org/10.1158/2159-8290. cd-15-0583.
[48] K. Reinhard et al., “An RNA vaccine drives expansion and efficacy of claudin-CAR-T cells against solid tumors,” Science, vol. 367, no. 6476, pp. 446–453, Jan. 2020, doi: https://doi.org/10.1126/ science.aay 5967.
[49] C. R. Stadler et al., “Preclinical efficacy and pharmacokinetics of an RNA-encoded T cell–engaging bispecific antibody targeting human claudin 6,” Science Translational Medicine, vol. 16, no. 748, May 2024, doi: https://doi.org/10.1126/scitranslmed.adl2720.
[50] J.-I. Youn and D. I. Gabrilovich, “The biology of myeloid-derived suppressor cells: The blessing and the curse of morphological and functional heterogeneity,” European Journal of Immunology, vol. 40, no. 11, pp. 2969–2975, 2010, doi: https://doi.org/10.1002/eji.201040895.
[51] O. S. Blomberg, L. Spagnuolo, and K. E. de Visser, “Immune regulation of metastasis: mechanistic insights and therapeutic opportunities,” Disease Models & Mechanisms, vol. 11, no. 10, 2018, doi: https://doi.org/10.1242/dmm.036236.
[52] K. Okła et al., “Clinical Relevance and Immunosuppressive Pattern of Circulating and Infiltrating Subsets of Myeloid-Derived Suppressor Cells (MDSCs) in Epithelial Ovarian Cancer,” Frontiers in Immunology, vol. 10, Apr. 2019, doi: https://doi.org/10.3389/fimmu.2019.00691.
[53] D. I. Gabrilovich, “Myeloid-Derived Suppressor Cells,” Cancer Immunology Research, vol. 5, no. 1, pp. 3–8, 2017, doi: https://doi.org/10.1158/2326-6066.cir-16-0297.
[54] S. Ruixin et al., “CXCR4-modified CAR-T cells suppresses MDSCs recruitment via STAT3/NF-κB/SDF-1α axis to enhance efficacy against pancreatic cancer,” Molecular Therapy, vol. 31, no. 11, pp. 3193–3209, Nov. 2023, doi: https://doi.org/10.1016/j.ymthe.2023.09.010.
[55] D. Raman, P. J. Baugher, Y. M. Thu, and A. Richmond, “Role of chemokines in tumor growth,” Cancer Letters, vol. 256, no. 2, pp. 137–165, 2007, doi: https://doi.org/10.1016/j.canlet.2007.05.013.
[56] K. Bullock and A. Richmond, “Suppressing MDSC Recruitment to the Tumor Microenvironment by Antagonizing CXCR2 to Enhance the Efficacy of Immunotherapy,” Cancers, vol. 13, no. 24, p. 6293, Dec. 2021, doi: https://doi.org/10.3390/cancers13246293.
[57] L. Jin et al., “CXCR1- or CXCR2-modified CAR T cells co-opt IL-8 for maximal antitumor efficacy in solid tumors,” Nature Communications, vol. 10, no. 1, Sep. 2019, doi: https://doi.org/10.1038/s41467-019-11869-4.
[58] L. M. Whilding et al., “CAR T-Cells Targeting the Integrin αvβ6 and Co-Expressing the Chemokine Receptor CXCR2 Demonstrate Enhanced Homing and Efficacy against Several Solid Malignancies,” Cancers, vol. 11, no. 5, p. 674, May 2019, doi: https://doi.org/10.3390/cancers11050674.
[59] G. Liu et al., “CXCR2-modified CAR-T cells have enhanced trafficking ability that improves treatment of hepatocellular carcinoma,” European Journal of Immunology, vol. 50, no. 5, pp. 712–724, May 2020, doi: https://doi.org/10.1002/eji.201948457.
[60] X. Zhang, R. Guo, H. Kambara, F. Ma, and H. Luo, “The role of CXCR2 in acute inflammatory responses and its antagonists as anti-inflammatory therapeutics,” Current Opinion in Hematology, vol. 26, no. 1, pp. 28–33, 2019, doi: https://doi.org/10.1097/moh.0000000000000476.
[61] L. J. Goldstein et al., “A window-of-opportunity trial of the CXCR1/2 inhibitor reparixin in operable HER-2-negative breast cancer,” Breast cancer research: BCR, vol. 22, no. 1, p. 4, Jan. 2020, doi: https://doi.org/10.1186/s13058-019-1243-8.
[62] S. Greene et al., “Inhibition of MDSC Trafficking with SX-682, a CXCR1/2 Inhibitor, Enhances NK-Cell Immunotherapy in Head and Neck Cancer Models,” Clinical Cancer Research: An Official Journal of the American Association for Cancer Research, vol. 26, no. 6, pp. 1420–1431, Mar. 2020, doi: https://doi.org/10.1158/1078-0432.CCR-19-2625.
[63] K. H. Tang et al., “Combined Inhibition of SHP2 and CXCR1/2 Promotes Antitumor T-cell Response in NSCLC,” Cancer discovery, vol. 12, no. 1, pp. 47–61, Aug. 2021, doi: https://doi.org/10.1158/2159-8290. cd-21-0369.
[64] J. Leslie et al., “CXCR2 inhibition enables NASH-HCC immunotherapy,” Gut, vol. 71, no. 10, pp. 2093–2106, Apr. 2022, doi: https://doi.org/10.1136/gutjnl-2021-326259.
[65] G. Lazennec, K. Rajarathnam, and A. Richmond, “CXCR2 chemokine receptor - a master regulator in cancer and physiology,” Trends in Molecular Medicine, vol. 30, no. 1, pp. 37–55, 2024, doi: https://doi. org/ 10.1016/j.molmed.2023.09.003.
[66] J. W. Kwak et al., “CXCR1/2 antagonism inhibits neutrophil function and not recruitment in cancer,” OncoImmunology, vol. 13, no. 1, Jul. 2024, doi: https://doi.org/10.1080/2162402x.2024.2384674.
[67] Y. Zhang et al., “Controlling T cell-tumor cell interaction with a biomimetic physical barrier for cancer immunotherapy.,” PubMed, vol. 122, no. 28, Jul. 2025, doi: https://doi.org/10.1073/pnas.2500589122.
[68] Caruana et al., “Heparanase promotes tumor infiltration and antitumor activity of CAR-redirected T lymphocytes,” Nature Medicine, vol. 21, no. 5, pp. 524–529, May 2015, doi: https://doi.org/10. 1038/nm. 3833.
[69] L.-R. Sofia et al., “An integral membrane constitutively active heparanase enhances the tumor infiltration capability of NK cells,” Oncoimmunology, vol. 14, no. 1, Dec. 2024, doi: https://doi.org/ 10.1080/ 2162402X.2024.2437917.
[70] H. A. Martinez et al., “Regulatory T cells use heparanase to access IL-2 bound to extracellular matrix in inflamed tissue,” Nature communications, vol. 15, no. 1, Feb. 2024, doi: https://doi.org/10.1038/s41467-024-45012-9.
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