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 evaluat...

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Main Authors: 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, Jan Molenaar, Jarno Drost, Anne C. Rios, Kerry Chester, Judith Wienke, John Anderson
Format: Article
Language:English
Published: Nature Portfolio 2025-08-01
Series:Nature Communications
Online Access:https://doi.org/10.1038/s41467-025-61427-4
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author 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
Jan Molenaar
Jarno Drost
Anne C. Rios
Kerry Chester
Judith Wienke
John Anderson
author_facet 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
Jan Molenaar
Jarno Drost
Anne C. Rios
Kerry Chester
Judith Wienke
John Anderson
author_sort Marta Barisa
collection DOAJ
description 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.
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spelling doaj-art-b7d2cd39dd0542bbae8cd1123c094baf2025-08-20T04:02:55ZengNature PortfolioNature Communications2041-17232025-08-0116112010.1038/s41467-025-61427-4Functional avidity of anti-B7H3 CAR-T constructs predicts antigen density thresholds for triggering effector functionMarta Barisa0Henrike P. Muller1Elisa Zappa2Rivani Shah3Juliane L. Buhl4Benjamin Draper5Courtney Himsworth6Chantelle Bowers7Sophie Munnings-Tomes8Marilena Nicolaidou9Sonia Morlando10Kathleen Birley11Clara Leboreiro-Babe12Alice Vitali13Laura Privitera14Kyle O’Sullivan15Ailsa Greppi16Magdalena Buschhaus17Mario Barrera Román18Sam de Blank19Femke van den Ham20Brenna R. van ‘t Veld21Gabrielle Ferry22Jonathan Fisher23Debarati Shome24Reza Nadafi25Israrul H. Ansari26Rogier Reijmers27Stefano Giuliani28Paul Sondel29Laura K. Donovan30Louis Chesler31Jan Molenaar32Jarno Drost33Anne C. Rios34Kerry Chester35Judith Wienke36John Anderson37Great Ormond Street Institute of Child Health, University College LondonGreat Ormond Street Institute of Child Health, University College LondonPrincess Máxima Center for Pediatric OncologyGreat Ormond Street Institute of Child Health, University College LondonPrincess Máxima Center for Pediatric OncologyGreat Ormond Street Institute of Child Health, University College LondonGreat Ormond Street Institute of Child Health, University College LondonGreat Ormond Street Institute of Child Health, University College LondonGreat Ormond Street Institute of Child Health, University College LondonGreat Ormond Street Institute of Child Health, University College LondonGreat Ormond Street Institute of Child Health, University College LondonGreat Ormond Street Institute of Child Health, University College LondonGreat Ormond Street Institute of Child Health, University College LondonGreat Ormond Street Institute of Child Health, University College LondonGreat Ormond Street Institute of Child Health, University College LondonGreat Ormond Street Institute of Child Health, University College LondonGreat Ormond Street Institute of Child Health, University College LondonLumicksPrincess Máxima Center for Pediatric OncologyPrincess Máxima Center for Pediatric OncologyPrincess Máxima Center for Pediatric OncologyPrincess Máxima Center for Pediatric OncologyGreat Ormond Street Institute of Child Health, University College LondonGreat Ormond Street Institute of Child Health, University College LondonLumicksLumicksDepartment of Pediatrics, University of Wisconsin School of Medicine and Public Health, University of Wisconsin-MadisonLumicksGreat Ormond Street Institute of Child Health, University College LondonDepartment of Pediatrics, University of Wisconsin School of Medicine and Public Health, University of Wisconsin-MadisonGreat Ormond Street Institute of Child Health, University College LondonCancer Institute, University College LondonPrincess Máxima Center for Pediatric OncologyPrincess Máxima Center for Pediatric OncologyPrincess Máxima Center for Pediatric OncologyThe Institute of Cancer ResearchPrincess Máxima Center for Pediatric OncologyGreat Ormond Street Institute of Child Health, University College LondonAbstract 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.https://doi.org/10.1038/s41467-025-61427-4
spellingShingle 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
Jan Molenaar
Jarno Drost
Anne C. Rios
Kerry Chester
Judith Wienke
John Anderson
Functional avidity of anti-B7H3 CAR-T constructs predicts antigen density thresholds for triggering effector function
Nature Communications
title Functional avidity of anti-B7H3 CAR-T constructs predicts antigen density thresholds for triggering effector function
title_full Functional avidity of anti-B7H3 CAR-T constructs predicts antigen density thresholds for triggering effector function
title_fullStr Functional avidity of anti-B7H3 CAR-T constructs predicts antigen density thresholds for triggering effector function
title_full_unstemmed Functional avidity of anti-B7H3 CAR-T constructs predicts antigen density thresholds for triggering effector function
title_short Functional avidity of anti-B7H3 CAR-T constructs predicts antigen density thresholds for triggering effector function
title_sort functional avidity of anti b7h3 car t constructs predicts antigen density thresholds for triggering effector function
url https://doi.org/10.1038/s41467-025-61427-4
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