Systemically administered low-affinity HER2 CAR T cells mediate antitumor efficacy without toxicity

Background The paucity of tumor-specific targets for chimeric antigen receptor (CAR) T-cell therapy of solid tumors necessitates careful preclinical evaluation of the therapeutic window for candidate antigens. Human epidermal growth factor receptor 2 (HER2) is an attractive candidate for CAR T-cell...

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Main Authors: Stanley R Riddell, Ekram A Gad, Sylvain Simon, Tamer Basel Shabaneh, Andrew R Stevens, Sylvia M Stull, Kristen R Shimp, Brandon W Seaton, Carla A Jaeger-Ruckstuhl, Amanda L Koehne, Jason P Price, James M Olson, Benjamin G Hoffstrom, David Jellyman
Format: Article
Language:English
Published: BMJ Publishing Group 2024-02-01
Series:Journal for ImmunoTherapy of Cancer
Online Access:https://jitc.bmj.com/content/12/2/e008566.full
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author Stanley R Riddell
Ekram A Gad
Sylvain Simon
Tamer Basel Shabaneh
Andrew R Stevens
Sylvia M Stull
Kristen R Shimp
Brandon W Seaton
Carla A Jaeger-Ruckstuhl
Amanda L Koehne
Jason P Price
James M Olson
Benjamin G Hoffstrom
David Jellyman
author_facet Stanley R Riddell
Ekram A Gad
Sylvain Simon
Tamer Basel Shabaneh
Andrew R Stevens
Sylvia M Stull
Kristen R Shimp
Brandon W Seaton
Carla A Jaeger-Ruckstuhl
Amanda L Koehne
Jason P Price
James M Olson
Benjamin G Hoffstrom
David Jellyman
author_sort Stanley R Riddell
collection DOAJ
description Background The paucity of tumor-specific targets for chimeric antigen receptor (CAR) T-cell therapy of solid tumors necessitates careful preclinical evaluation of the therapeutic window for candidate antigens. Human epidermal growth factor receptor 2 (HER2) is an attractive candidate for CAR T-cell therapy in humans but has the potential for eliciting on-target off-tumor toxicity. We developed an immunocompetent tumor model of CAR T-cell therapy targeting murine HER2 (mHER2) and examined the effect of CAR affinity, T-cell dose, and lymphodepletion on safety and efficacy.Methods Antibodies specific for mHER2 were generated, screened for affinity and specificity, tested for immunohistochemical staining of HER2 on normal tissues, and used for HER2-targeted CAR design. CAR candidates were evaluated for T-cell surface expression and the ability to induce T-cell proliferation, cytokine production, and cytotoxicity when transduced T cells were co-cultured with mHER2+ tumor cells in vitro. Safety and efficacy of various HER2 CARs was evaluated in two tumor models and normal non-tumor-bearing mice.Results Mice express HER2 in the same epithelial tissues as humans, rendering these tissues vulnerable to recognition by systemically administered HER2 CAR T cells. CAR T cells designed with single-chain variable fragment (scFvs) that have high-affinity for HER2 infiltrated and caused toxicity to normal HER2-positive tissues but exhibited poor infiltration into tumors and antitumor activity. In contrast, CAR T cells designed with an scFv with low-affinity for HER2 infiltrated HER2-positive tumors and controlled tumor growth without toxicity. Toxicity mediated by high-affinity CAR T cells was independent of tumor burden and correlated with proliferation of CAR T cells post infusion.Conclusions Our findings illustrate the disadvantage of high-affinity CARs for targets such as HER2 that are expressed on normal tissues. The use of low-affinity HER2 CARs can safely regress tumors identifying a potential path for therapy of solid tumors that exhibit high levels of HER2.
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spelling doaj.art-eef8e83670544a3da80201117fcb79112024-03-01T07:15:08ZengBMJ Publishing GroupJournal for ImmunoTherapy of Cancer2051-14262024-02-0112210.1136/jitc-2023-008566Systemically administered low-affinity HER2 CAR T cells mediate antitumor efficacy without toxicityStanley R Riddell0Ekram A Gad1Sylvain Simon2Tamer Basel Shabaneh3Andrew R Stevens4Sylvia M Stull5Kristen R Shimp6Brandon W Seaton7Carla A Jaeger-Ruckstuhl8Amanda L Koehne9Jason P Price10James M Olson11Benjamin G Hoffstrom12David Jellyman13Immunotherapy Integrated Research Center, Fred Hutchinson Cancer Center, Seattle, Washington, USAComparative Medicine, Fred Hutchinson Cancer Center, Seattle, Washington, USAImmunotherapy Integrated Research Center, Fred Hutchinson Cancer Center, Seattle, Washington, USAImmunotherapy Integrated Research Center, Fred Hutchinson Cancer Center, Seattle, Washington, USAImmunotherapy Integrated Research Center, Fred Hutchinson Cancer Center, Seattle, Washington, USAImmunotherapy Integrated Research Center, Fred Hutchinson Cancer Center, Seattle, Washington, USATranslational Science and Therapeutics Division, Fred Hutchinson Cancer Center, Seattle, Washington, USATranslational Science and Therapeutics Division, Fred Hutchinson Cancer Center, Seattle, Washington, USAImmunotherapy Integrated Research Center, Fred Hutchinson Cancer Center, Seattle, Washington, USAExperimental Histopathology, Fred Hutchinson Cancer Center, Seattle, Washington, USAMolecular Design and Therapeutics, Fred Hutchinson Cancer Center, Seattle, Washington, USAMolecular Design and Therapeutics, Fred Hutchinson Cancer Center, Seattle, Washington, USAAntibody Technology, Fred Hutchinson Cancer Center, Seattle, Washington, USAAntibody Technology, Fred Hutchinson Cancer Center, Seattle, Washington, USABackground The paucity of tumor-specific targets for chimeric antigen receptor (CAR) T-cell therapy of solid tumors necessitates careful preclinical evaluation of the therapeutic window for candidate antigens. Human epidermal growth factor receptor 2 (HER2) is an attractive candidate for CAR T-cell therapy in humans but has the potential for eliciting on-target off-tumor toxicity. We developed an immunocompetent tumor model of CAR T-cell therapy targeting murine HER2 (mHER2) and examined the effect of CAR affinity, T-cell dose, and lymphodepletion on safety and efficacy.Methods Antibodies specific for mHER2 were generated, screened for affinity and specificity, tested for immunohistochemical staining of HER2 on normal tissues, and used for HER2-targeted CAR design. CAR candidates were evaluated for T-cell surface expression and the ability to induce T-cell proliferation, cytokine production, and cytotoxicity when transduced T cells were co-cultured with mHER2+ tumor cells in vitro. Safety and efficacy of various HER2 CARs was evaluated in two tumor models and normal non-tumor-bearing mice.Results Mice express HER2 in the same epithelial tissues as humans, rendering these tissues vulnerable to recognition by systemically administered HER2 CAR T cells. CAR T cells designed with single-chain variable fragment (scFvs) that have high-affinity for HER2 infiltrated and caused toxicity to normal HER2-positive tissues but exhibited poor infiltration into tumors and antitumor activity. In contrast, CAR T cells designed with an scFv with low-affinity for HER2 infiltrated HER2-positive tumors and controlled tumor growth without toxicity. Toxicity mediated by high-affinity CAR T cells was independent of tumor burden and correlated with proliferation of CAR T cells post infusion.Conclusions Our findings illustrate the disadvantage of high-affinity CARs for targets such as HER2 that are expressed on normal tissues. The use of low-affinity HER2 CARs can safely regress tumors identifying a potential path for therapy of solid tumors that exhibit high levels of HER2.https://jitc.bmj.com/content/12/2/e008566.full
spellingShingle Stanley R Riddell
Ekram A Gad
Sylvain Simon
Tamer Basel Shabaneh
Andrew R Stevens
Sylvia M Stull
Kristen R Shimp
Brandon W Seaton
Carla A Jaeger-Ruckstuhl
Amanda L Koehne
Jason P Price
James M Olson
Benjamin G Hoffstrom
David Jellyman
Systemically administered low-affinity HER2 CAR T cells mediate antitumor efficacy without toxicity
Journal for ImmunoTherapy of Cancer
title Systemically administered low-affinity HER2 CAR T cells mediate antitumor efficacy without toxicity
title_full Systemically administered low-affinity HER2 CAR T cells mediate antitumor efficacy without toxicity
title_fullStr Systemically administered low-affinity HER2 CAR T cells mediate antitumor efficacy without toxicity
title_full_unstemmed Systemically administered low-affinity HER2 CAR T cells mediate antitumor efficacy without toxicity
title_short Systemically administered low-affinity HER2 CAR T cells mediate antitumor efficacy without toxicity
title_sort systemically administered low affinity her2 car t cells mediate antitumor efficacy without toxicity
url https://jitc.bmj.com/content/12/2/e008566.full
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