Functional avidity of anti-B7H3 CAR-T constructs predicts antigen density thresholds for triggering effector function

Marta Barisa; Henrike P. Muller; Elisa Zappa; Rivani Shah; Juliane L. Buhl; Benjamin Draper; Courtney Himsworth; Chantelle Bowers; Sophie Munnings-Tomes; Marilena Nicolaidou; Sonia Morlando; Kathleen Birley; Clara Leboreiro-Babe; Alice Vitali; Laura Privitera; Kyle O’Sullivan; Ailsa Greppi; Magdalena Buschhaus; Mario Barrera Román; Sam de Blank; Femke van den Ham; Brenna R. van ‘t Veld; Gabrielle Ferry; Jonathan Fisher; Debarati Shome; Reza Nadafi; Israrul H. Ansari; Rogier Reijmers; Stefano Giuliani; Paul Sondel; Laura K. Donovan; Louis Chesler; Molenaar Jan Molenaar; Jarno Drost; Anne C. Rios; Kerry Chester; Judith Wienke; John Anderson

doi:10.1038/s41467-025-61427-4

Functional avidity of anti-B7H3 CAR-T constructs predicts antigen density thresholds for triggering effector function

Marta Barisa
, Henrike P. Muller
, Elisa Zappa
, Rivani Shah
, Juliane L. Buhl
, Benjamin Draper
, Courtney Himsworth
, Chantelle Bowers
, Sophie Munnings-Tomes
, Marilena Nicolaidou
, Sonia Morlando
, Kathleen Birley
, Clara Leboreiro-Babe
, Alice Vitali
, Laura Privitera
, Kyle O’Sullivan
, Ailsa Greppi
, Magdalena Buschhaus
, Mario Barrera Román
, Sam de Blank

Femke van den Ham, Brenna R. van ‘t Veld, Gabrielle Ferry, Jonathan Fisher, Debarati Shome, Reza Nadafi, Israrul H. Ansari, Rogier Reijmers, Stefano Giuliani, Paul Sondel, Laura K. Donovan, Louis Chesler, Molenaar Jan Molenaar, Jarno Drost, Anne C. Rios, Kerry Chester, Judith Wienke, John Anderson

Research output: Contribution to journal › Article › peer-review

8 Citations (Scopus)

Abstract

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⁺ 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.

Original language	English
Article number	7196
Journal	Nature communications
Volume	16
Issue number	1
DOIs	https://doi.org/10.1038/s41467-025-61427-4
Publication status	Published - 5 Aug 2025

Keywords

Xenograft Model Antitumor Assays
CD8-Positive T-Lymphocytes/immunology
Animals
Immunotherapy, Adoptive/methods
Tumor Microenvironment/immunology
Humans
Cell Line, Tumor
Neoplasms/immunology
Female
Receptors, Chimeric Antigen/immunology
Mice
B7 Antigens/immunology

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10.1038/s41467-025-61427-4

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Barisa, M., Muller, H. P., Zappa, E., Shah, R., Buhl, J. L., Draper, B., Himsworth, C., Bowers, C., Munnings-Tomes, S., Nicolaidou, M., Morlando, S., Birley, K., Leboreiro-Babe, C., Vitali, A., Privitera, L., O’Sullivan, K., Greppi, A., Buschhaus, M., Román, M. B., ... Anderson, J. (2025). Functional avidity of anti-B7H3 CAR-T constructs predicts antigen density thresholds for triggering effector function. Nature communications, 16(1), Article 7196. https://doi.org/10.1038/s41467-025-61427-4

@article{7572b8da66df44cfb81908b56f48858d,

title = "Functional avidity of anti-B7H3 CAR-T constructs predicts antigen density thresholds for triggering effector function",

abstract = "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+ 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.",

keywords = "Xenograft Model Antitumor Assays, CD8-Positive T-Lymphocytes/immunology, Animals, Immunotherapy, Adoptive/methods, Tumor Microenvironment/immunology, Humans, Cell Line, Tumor, Neoplasms/immunology, Female, Receptors, Chimeric Antigen/immunology, Mice, B7 Antigens/immunology",

author = "Marta Barisa and Muller, \{Henrike P.\} and Elisa Zappa and Rivani Shah and Buhl, \{Juliane L.\} and Benjamin Draper and Courtney Himsworth and Chantelle Bowers and Sophie Munnings-Tomes and Marilena Nicolaidou and Sonia Morlando and Kathleen Birley and Clara Leboreiro-Babe and Alice Vitali and Laura Privitera and Kyle O{\textquoteright}Sullivan and Ailsa Greppi and Magdalena Buschhaus and Rom{\'a}n, \{Mario Barrera\} and \{de Blank\}, Sam and \{van den Ham\}, Femke and \{van {\textquoteleft}t Veld\}, \{Brenna R.\} and Gabrielle Ferry and Jonathan Fisher and Debarati Shome and Reza Nadafi and Ansari, \{Israrul H.\} and Rogier Reijmers and Stefano Giuliani and Paul Sondel and Donovan, \{Laura K.\} and Louis Chesler and \{Jan Molenaar\}, Molenaar and Jarno Drost and Rios, \{Anne C.\} and Kerry Chester and Judith Wienke and John Anderson",

note = "{\textcopyright} 2025. The Author(s).",

year = "2025",

month = aug,

day = "5",

doi = "10.1038/s41467-025-61427-4",

language = "English",

volume = "16",

journal = "Nature communications",

issn = "2041-1723",

publisher = "Nature Research",

number = "1",

}

Barisa, M, Muller, HP, Zappa, E, Shah, R, Buhl, JL, Draper, B, Himsworth, C, Bowers, C, Munnings-Tomes, S, Nicolaidou, M, Morlando, S, Birley, K, Leboreiro-Babe, C, Vitali, A, Privitera, L, O’Sullivan, K, Greppi, A, Buschhaus, M, Román, MB, de Blank, S, van den Ham, F, van ‘t Veld, BR, Ferry, G, Fisher, J, Shome, D, Nadafi, R, Ansari, IH, Reijmers, R, Giuliani, S, Sondel, P, Donovan, LK, Chesler, L, Jan Molenaar, M, Drost, J , Rios, AC, Chester, K, Wienke, J & Anderson, J 2025, 'Functional avidity of anti-B7H3 CAR-T constructs predicts antigen density thresholds for triggering effector function', Nature communications, vol. 16, no. 1, 7196. https://doi.org/10.1038/s41467-025-61427-4

TY - JOUR

T1 - Functional avidity of anti-B7H3 CAR-T constructs predicts antigen density thresholds for triggering effector function

AU - Barisa, Marta

AU - Muller, Henrike P.

AU - Zappa, Elisa

AU - Shah, Rivani

AU - Buhl, Juliane L.

AU - Draper, Benjamin

AU - Himsworth, Courtney

AU - Bowers, Chantelle

AU - Munnings-Tomes, Sophie

AU - Nicolaidou, Marilena

AU - Morlando, Sonia

AU - Birley, Kathleen

AU - Leboreiro-Babe, Clara

AU - Vitali, Alice

AU - Privitera, Laura

AU - O’Sullivan, Kyle

AU - Greppi, Ailsa

AU - Buschhaus, Magdalena

AU - Román, Mario Barrera

AU - de Blank, Sam

AU - van den Ham, Femke

AU - van ‘t Veld, Brenna R.

AU - Ferry, Gabrielle

AU - Fisher, Jonathan

AU - Shome, Debarati

AU - Nadafi, Reza

AU - Ansari, Israrul H.

AU - Reijmers, Rogier

AU - Giuliani, Stefano

AU - Sondel, Paul

AU - Donovan, Laura K.

AU - Chesler, Louis

AU - Jan Molenaar, Molenaar

AU - Drost, Jarno

AU - Rios, Anne C.

AU - Chester, Kerry

AU - Wienke, Judith

AU - Anderson, John

PY - 2025/8/5

Y1 - 2025/8/5

N2 - 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+ 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.

AB - 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+ 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.

KW - Xenograft Model Antitumor Assays

KW - CD8-Positive T-Lymphocytes/immunology

KW - Animals

KW - Immunotherapy, Adoptive/methods

KW - Tumor Microenvironment/immunology

KW - Humans

KW - Cell Line, Tumor

KW - Neoplasms/immunology

KW - Female

KW - Receptors, Chimeric Antigen/immunology

KW - Mice

KW - B7 Antigens/immunology

UR - https://www.scopus.com/pages/publications/105012634359

UR - https://www.mendeley.com/catalogue/4dd84fb3-b518-3f36-9caf-640f7e390a44/

U2 - 10.1038/s41467-025-61427-4

DO - 10.1038/s41467-025-61427-4

M3 - Article

C2 - 40764385

AN - SCOPUS:105012634359

SN - 2041-1723

VL - 16

JO - Nature communications

JF - Nature communications

IS - 1

M1 - 7196

ER -

Functional avidity of anti-B7H3 CAR-T constructs predicts antigen density thresholds for triggering effector function

Abstract

Keywords

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