Dual-target CAR-T therapy for ovarian cancer: synergistic targeting of MSLN and B7H3 enhances anti-tumor efficacy and overcomes antigen heterogeneity

Dual-target CAR-T therapy for ovarian cancer: synergistic targeting of MSLN and B7H3 enhances anti-tumor efficacy and overcomes antigen heterogeneity

  • Bray F, Ferlay J, Soerjomataram I, Siegel RL, Torre LA, Jemal A. Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin. 2018;68:394–424.

    PubMed 

    Google Scholar
     

  • Torre LA, Trabert B, DeSantis CE, Miller KD, Samimi G, Runowicz CD, et al. Ovarian cancer statistics, 2018. CA Cancer J Clin. 2018;68:284–96.

    PubMed 
    PubMed Central 

    Google Scholar
     

  • Bouchkouj N, Kasamon YL, de Claro RA, George B, Lin X, Lee S, et al. FDA approval summary: axicabtagene ciloleucel for relapsed or refractory large b-cell lymphoma. Clin Cancer Res. 2019;25:1702–8.

    Article 
    PubMed 

    Google Scholar
     

  • Munshi NC, Anderson LDJ, Shah N, Madduri D, Berdeja J, Lonial S, et al. Idecabtagene vicleucel in relapsed and refractory multiple myeloma. N Engl J Med. 2021;384:705–16.

    Article 
    CAS 
    PubMed 

    Google Scholar
     

  • Wagner J, Wickman E, DeRenzo C, Gottschalk S. CAR t cell therapy for solid tumors: bright future or dark reality?. Mol Ther. 2020;28:2320–39.

    Article 
    CAS 
    PubMed 
    PubMed Central 

    Google Scholar
     

  • Marofi F, Motavalli R, Safonov VA, Thangavelu L, Yumashev AV, Alexander M, et al. CAR t cells in solid tumors: challenges and opportunities. Stem Cell Res Ther. 2021;12:81.

    Article 
    CAS 
    PubMed 
    PubMed Central 

    Google Scholar
     

  • Klampatsa A, Dimou V, Albelda SM. Mesothelin-targeted CAR-t cell therapy for solid tumors. Expert Opin Biol Ther. 2021;21:473–86.

    Article 
    CAS 
    PubMed 

    Google Scholar
     

  • Scales SJ, Gupta N, Pacheco G, Firestein R, French DM, Koeppen H, et al. An antimesothelin-monomethyl auristatin e conjugate with potent antitumor activity in ovarian, pancreatic, and mesothelioma models. Mol Cancer Ther. 2014;13:2630–40.

    Article 
    CAS 
    PubMed 

    Google Scholar
     

  • Winter JM, Tang LH, Klimstra DS, Brennan MF, Brody JR, Rocha FG, et al. A novel survival-based tissue microarray of pancreatic cancer validates MUC1 and mesothelin as biomarkers. PLoS ONE. 2012;7:e40157.

    Article 
    CAS 
    PubMed 
    PubMed Central 

    Google Scholar
     

  • Ho M, Bera TK, Willingham MC, Onda M, Hassan R, FitzGerald D, et al. Mesothelin expression in human lung cancer. Clin Cancer Res. 2007;13:1571–5.

    Article 
    CAS 
    PubMed 

    Google Scholar
     

  • Baba K, Ishigami S, Arigami T, Uenosono Y, Okumura H, Matsumoto M, et al. Mesothelin expression correlates with prolonged patient survival in gastric cancer. J Surg Oncol. 2012;105:195–9.

    Article 
    CAS 
    PubMed 

    Google Scholar
     

  • Obulhasim G, Fujii H, Matsumoto T, Yasen M, Abe M, Matsuoka S, et al. Mesothelin gene expression and promoter methylation/hypomethylation in gynecological tumors. Eur J Gynaecol Oncol. 2010;31:63–71.

    CAS 
    PubMed 

    Google Scholar
     

  • Tozbikian G, Brogi E, Kadota K, Catalano J, Akram M, Patil S, et al. Mesothelin expression in triple negative breast carcinomas correlates significantly with basal-like phenotype, distant metastases and decreased survival. PLoS One. 2014;9:e114900.

    Article 
    PubMed 
    PubMed Central 

    Google Scholar
     

  • Zhao B, Li H, Xia Y, Wang Y, Wang Y, Shi Y, et al. Immune checkpoint of b7-h3 in cancer: from immunology to clinical immunotherapy. J Hematol Oncol. 2022;15:153.

    Article 
    CAS 
    PubMed 
    PubMed Central 

    Google Scholar
     

  • Majzner RG, Theruvath JL, Nellan A, Heitzeneder S, Cui Y, Mount CW, et al. CAR t cells targeting b7-h3, a pan-cancer antigen, demonstrate potent preclinical activity against pediatric solid tumors and brain tumors. Clin Cancer Res. 2019;25:2560–74.

    Article 
    CAS 
    PubMed 
    PubMed Central 

    Google Scholar
     

  • Beatty GL, O’Hara MH, Lacey SF, Torigian DA, Nazimuddin F, Chen F, et al. Activity of mesothelin-specific chimeric antigen receptor t cells against pancreatic carcinoma metastases in a phase 1 trial. Gastroenterology. 2018;155:29–32.

    Article 
    CAS 
    PubMed 

    Google Scholar
     

  • Vitanza NA, Wilson AL, Huang W, Seidel K, Brown C, Gustafson JA, et al. Intraventricular b7-h3 CAR t cells for diffuse intrinsic pontine glioma: preliminary first-in-human bioactivity and safety. Cancer Discov. 2023;13:114–31.

    Article 
    CAS 
    PubMed 

    Google Scholar
     

  • Saif WM, Cassier PA, Curigliano G, Daniele G, Hilton JF, Koganemaru S, et al. A phase 1 dose escalation/expansion study of GSK5764227 (GSK’227), a b7-homolog 3 (b7-h3) protein targeted antibody-drug conjugate (ADC), in patients with advanced solid tumors, including gastrointestinal (GI) cancers. J Clin Oncol. 2025;43:TPS84.

    Article 

    Google Scholar
     

  • Zhang E, Yang P, Gu J, Wu H, Chi X, Liu C, et al. Recombination of a dual-CAR-modified t lymphocyte to accurately eliminate pancreatic malignancy. J Hematol Oncol. 2018;11:102.

    Article 
    PubMed 
    PubMed Central 

    Google Scholar
     

  • Yin Y, Rodriguez JL, Li N, Thokala R, Nasrallah MP, Hu L, et al. Locally secreted BiTEs complement CAR t cells by enhancing killing of antigen heterogeneous solid tumors. Mol Ther. 2022;30:2537–53.

    Article 
    CAS 
    PubMed 
    PubMed Central 

    Google Scholar
     

  • Hegde M, Mukherjee M, Grada Z, Pignata A, Landi D, Navai SA, et al. Tandem CAR t cells targeting HER2 and IL13ralpha2 mitigate tumor antigen escape. J Clin Invest. 2016;126:3036–52.

    Article 
    PubMed 
    PubMed Central 

    Google Scholar
     

  • Choi BD, Yu X, Castano AP, Bouffard AA, Schmidts A, Larson RC, et al. CAR-t cells secreting BiTEs circumvent antigen escape without detectable toxicity. Nat Biotechnol. 2019;37:1049–58.

    Article 
    CAS 
    PubMed 

    Google Scholar
     

  • Lanitis E, Poussin M, Klattenhoff AW, Song D, Sandaltzopoulos R, June CH, et al. Chimeric antigen receptor t cells with dissociated signaling domains exhibit focused antitumor activity with reduced potential for toxicity in vivo. Cancer Immunol Res. 2013;1:43–53.

    Article 
    CAS 
    PubMed 
    PubMed Central 

    Google Scholar
     

  • Ji F, Yu J, Zhu Y, Lin H, Gao K, Rao M, et al. B4m3 CAR-t cell enhance antitumor activity and non-tumor toxicity in ovarian cancer. Int Immunopharmacol. 2025;159:114919.

    Article 
    CAS 
    PubMed 

    Google Scholar
     

  • Ji F, Xu L, Long K, Zhang F, Zhang M, Lu X, et al. Rabies virus glycoprotein 29 (RVG29) promotes CAR-t immunotherapy for glioma. Transl Res. 2023;259:1–12.

    Article 
    CAS 
    PubMed 

    Google Scholar
     

  • Park AK, Fong Y, Kim S, Yang J, Murad JP, Lu J, et al. Effective combination immunotherapy using oncolytic viruses to deliver CAR targets to solid tumors. Sci Transl Med. 2020;12:eaaz1863.

  • Zhai Y, Du Y, Li G, Yu M, Hu H, Pan C, et al. Trogocytosis of CAR molecule regulates CAR-t cell dysfunction and tumor antigen escape. Signal Transduct Target Ther. 2023;8:457.

    Article 
    CAS 
    PubMed 
    PubMed Central 

    Google Scholar
     

  • Rodriguez-Marquez P, Calleja-Cervantes ME, Serrano G, Oliver-Caldes A, Palacios-Berraquero ML, Martin-Mallo A, et al. CAR density influences antitumoral efficacy of BCMA CAR t cells and correlates with clinical outcome. Sci Adv. 2022;8:eabo0514.

    Article 
    CAS 
    PubMed 
    PubMed Central 

    Google Scholar
     

  • Ruella M, Korell F, Porazzi P, Maus MV. Mechanisms of resistance to chimeric antigen receptor-t cells in haematological malignancies. Nat Rev Drug Discov. 2023;22:976–95.

    Article 
    CAS 
    PubMed 
    PubMed Central 

    Google Scholar
     

  • Hu Y, Hou J, Jiang Z, Lin Q. Mechanisms of resistance to CAR-t cell therapy in multiple myeloma: latest updates from the 2024 ASH annual meeting. Exp Hematol Oncol. 2025;14:45.

    Article 
    PubMed 
    PubMed Central 

    Google Scholar
     

  • Marchal I. FOXO1 enhances CAR t cell fitness and function. Nat Biotechnol. 2024;42:699.

    PubMed 

    Google Scholar
     

  • Stewart C, Ralyea C, Lockwood S. Ovarian cancer: an integrated review. Semin Oncol Nurs. 2019;35:151–6.

    Article 
    PubMed 

    Google Scholar
     

  • Bu L, Yan J, Wang Z, Ruan H, Chen Q, Gunadhi V, et al. Advances in drug delivery for post-surgical cancer treatment. Biomaterials. 2019;219:119182.

    Article 
    CAS 
    PubMed 

    Google Scholar
     

  • Sommer C, Boldajipour B, Kuo TC, Bentley T, Sutton J, Chen A, et al. Preclinical evaluation of allogeneic CAR t cells targeting BCMA for the treatment of multiple myeloma. Mol Ther. 2019;27:1126–38.

    Article 
    CAS 
    PubMed 
    PubMed Central 

    Google Scholar
     

  • Flugel CL, Majzner RG, Krenciute G, Dotti G, Riddell SR, Wagner DL, et al. Overcoming on-target, off-tumour toxicity of CAR t cell therapy for solid tumours. Nat Rev Clin Oncol. 2023;20:49–62.

    Article 
    CAS 
    PubMed 

    Google Scholar
     

  • Wang Y, Buck A, Piel B, Zerefa L, Murugan N, Coherd CD, et al. Affinity fine-tuning anti-CAIX CAR-t cells mitigate on-target off-tumor side effects. Mol Cancer. 2024;23:56.

    Article 
    CAS 
    PubMed 
    PubMed Central 

    Google Scholar
     

  • Majzner RG, Rietberg SP, Sotillo E, Dong R, Vachharajani VT, Labanieh L, et al. Tuning the antigen density requirement for CAR t-cell activity. Cancer Discov. 2020;10:702–23.

    Article 
    CAS 
    PubMed 
    PubMed Central 

    Google Scholar
     

  • Fry TJ, Shah NN, Orentas RJ, Stetler-Stevenson M, Yuan CM, Ramakrishna S, et al. CD22-targeted CAR t cells induce remission in b-ALL that is naive or resistant to CD19-targeted CAR immunotherapy. Nat Med. 2018;24:20–8.

    Article 
    CAS 
    PubMed 

    Google Scholar
     

  • Shi X, Yang J, Deng S, Xu H, Wu D, Zeng Q, et al. TGF-beta signaling in the tumor metabolic microenvironment and targeted therapies. J Hematol Oncol. 2022;15:135.

    Article 
    PubMed 
    PubMed Central 

    Google Scholar
     

  • Batlle E, Massague J. Transforming growth factor-beta signaling in immunity and cancer. Immunity. 2019;50:924–40.

    Article 
    CAS 
    PubMed 
    PubMed Central 

    Google Scholar
     

  • Lan Y, Moustafa M, Knoll M, Xu C, Furkel J, Lazorchak A, et al. Simultaneous targeting of TGF-beta/PD-l1 synergizes with radiotherapy by reprogramming the tumor microenvironment to overcome immune evasion. Cancer Cell. 2021;39:1388–403.

    Article 
    CAS 
    PubMed 

    Google Scholar
     

  • Doan AE, Mueller KP, Chen AY, Rouin GT, Chen Y, Daniel B, et al. FOXO1 is a master regulator of memory programming in CAR t cells. Nature. 2024;629:211–8.

    Article 
    CAS 
    PubMed 
    PubMed Central 

    Google Scholar
     

  • Carlson CM, Endrizzi BT, Wu J, Ding X, Weinreich MA, Walsh ER, et al. Kruppel-like factor 2 regulates thymocyte and t-cell migration. Nature. 2006;442:299–302.

    Article 
    CAS 
    PubMed 

    Google Scholar
     

  • Jung YW, Kim HG, Perry CJ, Kaech SM. CCR7 expression alters memory CD8 t-cell homeostasis by regulating occupancy in IL-7- and IL-15-dependent niches. Proc Natl Acad Sci USA. 2016;113:8278–83.

    Article 
    CAS 
    PubMed 
    PubMed Central 

    Google Scholar
     

  • Dominguez CX, Muller S, Keerthivasan S, Koeppen H, Hung J, Gierke S, et al. Single-cell RNA sequencing reveals stromal evolution into LRRC15(+) myofibroblasts as a determinant of patient response to cancer immunotherapy. Cancer Discov. 2020;10:232–53.

    Article 
    CAS 
    PubMed 

    Google Scholar
     

  • Levstek L, Janzic L, Ihan A, Kopitar AN. Biomarkers for prediction of CAR t therapy outcomes: current and future perspectives. Front Immunol. 2024;15:1378944.

    Article 
    CAS 
    PubMed 
    PubMed Central 

    Google Scholar
     

  • Garcia-Murillas I, Schiavon G, Weigelt B, Ng C, Hrebien S, Cutts RJ, et al. Mutation tracking in circulating tumor DNA predicts relapse in early breast cancer. Sci Transl Med. 2015;7:302ra133.

    Article 
    PubMed 

    Google Scholar
     

  • Li S, Lai H, Liu J, Liu Y, Jin L, Li Y, et al. Circulating tumor DNA predicts the response and prognosis in patients with early breast cancer receiving neoadjuvant chemotherapy. JCO Precis Oncol. 2020;4:PO.19.00292.

  • Hope JL, Stairiker CJ, Bae E, Otero DC, Bradley LM. Striking a balance-cellular and molecular drivers of memory t cell development and responses to chronic stimulation. Front Immunol. 2019;10:1595.

    Article 
    CAS 
    PubMed 
    PubMed Central 

    Google Scholar
     

  • Michelini Hess, Doedens R, Goldrath AL, Hedrick AW. SM. Differentiation of CD8 memory t cells depends on foxo1. J Exp Med. 2013;210:1189–200.

    Article 
    CAS 
    PubMed Central 

    Google Scholar
     

  • Zhang H, Yang Z, Yuan W, Liu J, Luo X, Zhang Q, et al. Sustained AhR activity programs memory fate of early effector CD8(+) t cells. Proc Natl Acad Sci USA. 2024;121:e2317658121.

    Article 
    PubMed 
    PubMed Central 

    Google Scholar
     

  • Agliardi G, Dias J, Rampotas A, Garcia J, Roddie C. Accelerating and optimising CAR t-cell manufacture to deliver better patient products. Lancet Haematol. 2025;12:e57–67.

    Article 
    CAS 
    PubMed 

    Google Scholar
     

  • Rupp LJ, Schumann K, Roybal KT, Gate RE, Ye CJ, Lim WA, et al. CRISPR/cas9-mediated PD-1 disruption enhances anti-tumor efficacy of human chimeric antigen receptor t cells. Sci Rep. 2017;7:737.

    Article 
    PubMed 
    PubMed Central 

    Google Scholar