Functional avidity of anti-B7H3 CAR-T constructs predicts antigen density thresholds for triggering effector function
Chimeric Antigen receptor T cell (CAR-T) treatments for solid cancers have been compromised by limited expansion and survival in the tumor microenvironment following interaction with antigen-expressing target cells. Using B7H3 as a model antigen with broad clinical applicability, we evaluate the relationship between the antibody/antigen affinity of three clinical candidate binders and the three following characteristics: cellular avidity, duration of sustained cytotoxicity in tumoroid re-stimulation assays, and in vivo anti-tumoral responses. Next, BEHAV3D video microscopy is used to assess CAR-T cell interaction with tumor cells at single cell resolution. These data are consistent with a threshold avidity of CAR-T / tumor cell interaction and target cell B7H3 expression level, where enhanced functionality is characterized by longer cumulative CD8<sup>+</sup> CAR-T / tumor target interaction times, CAR-T cell expansion and sustained tumor control. Lower checkpoint receptor expression does not correlate with enhanced anti-tumor function. These results provide further insights into design of anti-B7H3 CAR-T cells for antigen-dim cell targeting, and avoidance of antigen-dim tumor relapse.
Authors
- John A. Anderson (ORCID: https://orcid.org/0000-0001-7509-3203)
- Marta Barisa (ORCID: https://orcid.org/0000-0001-7846-6810)
- Clara Leboreiro-Babe (ORCID: https://orcid.org/0000-0003-4630-095X)
- Rogier M. Reijmers (ORCID: https://orcid.org/0000-0001-7471-7715)
- Chantelle Bowers (ORCID: https://orcid.org/0009-0006-7765-5891)
- Paul M. Sondel (ORCID: https://orcid.org/0000-0002-0981-8875)
- Juliane L. Buhl (ORCID: https://orcid.org/0000-0001-6505-3769)
- Kathleen Birley
- Sonia Morlando (ORCID: https://orcid.org/0000-0002-9010-1336)
- Magdalena Buschhaus (ORCID: https://orcid.org/0000-0002-5705-8997)
- Debarati Shome (ORCID: https://orcid.org/0000-0001-5992-883X)
- Henrike Muller (ORCID: https://orcid.org/0000-0002-4682-4553)
- Laura Privitera (ORCID: https://orcid.org/0000-0002-6384-6367)
- Judith Wienke (ORCID: https://orcid.org/0000-0002-3835-6374)
- Sophie Munnings-Tomes
- Jarno Drost (ORCID: https://orcid.org/0000-0002-2941-6179)
- Israr-ul H. Ansari (ORCID: https://orcid.org/0000-0003-0326-0329)
- Reza Nadafi (ORCID: https://orcid.org/0000-0002-6769-4145)
- Jonathan Fisher (ORCID: https://orcid.org/0000-0003-3302-2241)
- Sam de Blank (ORCID: https://orcid.org/0000-0003-2191-5675)
- Laura Donovan (ORCID: https://orcid.org/0000-0001-5557-5539)
- Alice Vitali (ORCID: https://orcid.org/0009-0004-4375-0044)
- Marilena Nicolaidou (ORCID: https://orcid.org/0009-0001-5306-8960)
- Mario Barrera Román (ORCID: https://orcid.org/0000-0002-5919-2962)
- Louis Chesler (ORCID: https://orcid.org/0000-0001-7842-2068)
- Kerry Chester (ORCID: https://orcid.org/0000-0002-3093-3130)
- Jan J. Molenaar (ORCID: https://orcid.org/0000-0002-2007-0431)
- Rivani Shah
- Stefano Giuliani (ORCID: https://orcid.org/0000-0002-4555-3093)
- Gabrielle M. Ferry (ORCID: https://orcid.org/0000-0002-0203-9326)
- Femke van den Ham
- Anne C. Rios (ORCID: https://orcid.org/0000-0002-9082-8068)
- Benjamin Draper (ORCID: https://orcid.org/0000-0001-7735-3988)
- Elisa Zappa
- Brenna R. van ‘t Veld
- Kyle O’Sullivan
- Ailsa Greppi
- Courtney Himsworth (ORCID: https://orcid.org/0000-0002-4233-7512)
Institutions
- University of Wisconsin–Madison (US)
- Institute of Cancer Research (GB)
- Great Ormond Street Hospital (GB)
- Oncode Institute (NL)
- Princess Máxima Center (NL)
- University College London (GB)
Publication Details
- Journal
- Nature Communications
- Published
- 2025-08-05
- DOI
- https://doi.org/10.1038/s41467-025-61427-4
- Citations
- 8
- Primary Topic
- CAR-T cell therapy research
- Type
- article
- Field-Weighted Citation Impact
- 8.36
Funders
- National Institute for Health and Care Research
- Horizon 2020 Framework Programme