T-cell receptors identified by a personalized antigen-agnostic screening approach target shared neoantigen KRAS Q61H

Adoptive cell therapy (ACT) with TCR-engineered T-cells represents a promising alternative to TIL- or CAR-T therapies for patients with advanced solid cancers. Currently, selection of therapeutic TCRs critically depends on knowing the target antigens, a condition excluding most patients from treatme...

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Main Authors: Volker Lennerz, Christoph Doppler, Martina Fatho, Anja Dröge, Sigrid Schaper, Kristin Gennermann, Nadine Genzel, Stephanie Plassmann, David Weismann, Samuel W. Lukowski, Dominik Bents, Christina Beushausen, Karen Kriese, Hermann Herbst, Volkhard Seitz, Rudolf Hammer, Paul J. Adam, Stephan Eggeling, Catherine Wölfel, Thomas Wölfel, Steffen Hennig
Format: Article
Language:English
Published: Frontiers Media S.A. 2025-03-01
Series:Frontiers in Immunology
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Online Access:https://www.frontiersin.org/articles/10.3389/fimmu.2025.1509855/full
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author Volker Lennerz
Volker Lennerz
Volker Lennerz
Christoph Doppler
Martina Fatho
Anja Dröge
Sigrid Schaper
Kristin Gennermann
Nadine Genzel
Stephanie Plassmann
David Weismann
Samuel W. Lukowski
Dominik Bents
Christina Beushausen
Karen Kriese
Hermann Herbst
Volkhard Seitz
Rudolf Hammer
Rudolf Hammer
Paul J. Adam
Stephan Eggeling
Catherine Wölfel
Thomas Wölfel
Steffen Hennig
Steffen Hennig
author_facet Volker Lennerz
Volker Lennerz
Volker Lennerz
Christoph Doppler
Martina Fatho
Anja Dröge
Sigrid Schaper
Kristin Gennermann
Nadine Genzel
Stephanie Plassmann
David Weismann
Samuel W. Lukowski
Dominik Bents
Christina Beushausen
Karen Kriese
Hermann Herbst
Volkhard Seitz
Rudolf Hammer
Rudolf Hammer
Paul J. Adam
Stephan Eggeling
Catherine Wölfel
Thomas Wölfel
Steffen Hennig
Steffen Hennig
author_sort Volker Lennerz
collection DOAJ
description Adoptive cell therapy (ACT) with TCR-engineered T-cells represents a promising alternative to TIL- or CAR-T therapies for patients with advanced solid cancers. Currently, selection of therapeutic TCRs critically depends on knowing the target antigens, a condition excluding most patients from treatment. Direct antigen-agnostic identification of tumor-specific T-cell clonotypes and TCR-T manufacturing using their TCRs can advance ACT for patients with aggressive solid cancers. We present a method to identify tumor-specific clonotypes from surgical specimens by comparing TCRβ-chain repertoires of TILs and adjacent tissue-resident lymphocytes. In six out of seven NSCLC-patients analyzed, our selection of tumor-specific clonotypes based on TIL-abundance and high tumor-to-nontumor frequency ratios was confirmed by gene expression signatures determined by scRNA-Seq. In three patients, we demonstrated that predicted tumor-specific clonotypes reacted against autologous tumors. For one of these patients, we engineered TCR-T cells with four candidate tumor-specific TCRs that showed reactivity against the patient’s tumor and HLA-matched NSCLC cell lines. The TCR-T cells were then used to screen for candidate neoantigens and aberrantly expressed antigens. Three TCRs recognized recurrent driver-mutation KRAS Q61H-peptide ILDTAGHEEY presented by HLA-A*01:01. The TCRs were also dominant in a tumor relapse, one was found in cell free DNA. The finding of homologous TCRs in independent KRAS Q61H-positive cancers suggests a therapeutic opportunity for HLA-matched patients with KRAS Q61H-expressing tumors.
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spelling doaj-art-8b93b9403ffa44d5ac3fee587f923cec2025-08-20T02:56:02ZengFrontiers Media S.A.Frontiers in Immunology1664-32242025-03-011610.3389/fimmu.2025.15098551509855T-cell receptors identified by a personalized antigen-agnostic screening approach target shared neoantigen KRAS Q61HVolker Lennerz0Volker Lennerz1Volker Lennerz2Christoph Doppler3Martina Fatho4Anja Dröge5Sigrid Schaper6Kristin Gennermann7Nadine Genzel8Stephanie Plassmann9David Weismann10Samuel W. Lukowski11Dominik Bents12Christina Beushausen13Karen Kriese14Hermann Herbst15Volkhard Seitz16Rudolf Hammer17Rudolf Hammer18Paul J. Adam19Stephan Eggeling20Catherine Wölfel21Thomas Wölfel22Steffen Hennig23Steffen Hennig24Internal Medicine III, University Medical Center (UMC) of the Johannes Gutenberg University Mainz, Mainz, GermanyHSDiagnomics GmbH, Berlin, GermanyTheryCell GmbH, Berlin, GermanyInternal Medicine III, University Medical Center (UMC) of the Johannes Gutenberg University Mainz, Mainz, GermanyInternal Medicine III, University Medical Center (UMC) of the Johannes Gutenberg University Mainz, Mainz, GermanyHSDiagnomics GmbH, Berlin, GermanyHSDiagnomics GmbH, Berlin, GermanyHSDiagnomics GmbH, Berlin, GermanyTheryCell GmbH, Berlin, GermanyTheryCell GmbH, Berlin, GermanyBoehringer Ingelheim RCV, GmbH & Co KG., Cancer Immunology & Immune Modulation, Vienna, AustriaBoehringer Ingelheim RCV, GmbH & Co KG., Cancer Immunology & Immune Modulation, Vienna, AustriaTheryCell GmbH, Berlin, GermanyVivantes Clinic Neukölln, Vivantes Thoracic Surgery, Berlin, GermanyVivantes Pathology, Vivantes Clinic Neukölln, Berlin, GermanyVivantes Pathology, Vivantes Clinic Neukölln, Berlin, GermanyHSDiagnomics GmbH, Berlin, GermanyHSDiagnomics GmbH, Berlin, GermanyTheryCell GmbH, Berlin, GermanyBoehringer Ingelheim RCV, GmbH & Co KG., Cancer Immunology & Immune Modulation, Vienna, AustriaVivantes Clinic Neukölln, Vivantes Thoracic Surgery, Berlin, GermanyInternal Medicine III, University Medical Center (UMC) of the Johannes Gutenberg University Mainz, Mainz, GermanyInternal Medicine III, University Medical Center (UMC) of the Johannes Gutenberg University Mainz, Mainz, GermanyHSDiagnomics GmbH, Berlin, GermanyTheryCell GmbH, Berlin, GermanyAdoptive cell therapy (ACT) with TCR-engineered T-cells represents a promising alternative to TIL- or CAR-T therapies for patients with advanced solid cancers. Currently, selection of therapeutic TCRs critically depends on knowing the target antigens, a condition excluding most patients from treatment. Direct antigen-agnostic identification of tumor-specific T-cell clonotypes and TCR-T manufacturing using their TCRs can advance ACT for patients with aggressive solid cancers. We present a method to identify tumor-specific clonotypes from surgical specimens by comparing TCRβ-chain repertoires of TILs and adjacent tissue-resident lymphocytes. In six out of seven NSCLC-patients analyzed, our selection of tumor-specific clonotypes based on TIL-abundance and high tumor-to-nontumor frequency ratios was confirmed by gene expression signatures determined by scRNA-Seq. In three patients, we demonstrated that predicted tumor-specific clonotypes reacted against autologous tumors. For one of these patients, we engineered TCR-T cells with four candidate tumor-specific TCRs that showed reactivity against the patient’s tumor and HLA-matched NSCLC cell lines. The TCR-T cells were then used to screen for candidate neoantigens and aberrantly expressed antigens. Three TCRs recognized recurrent driver-mutation KRAS Q61H-peptide ILDTAGHEEY presented by HLA-A*01:01. The TCRs were also dominant in a tumor relapse, one was found in cell free DNA. The finding of homologous TCRs in independent KRAS Q61H-positive cancers suggests a therapeutic opportunity for HLA-matched patients with KRAS Q61H-expressing tumors.https://www.frontiersin.org/articles/10.3389/fimmu.2025.1509855/fullT-cell receptor (TCR)TCR-T celltumor-specific antigenneoantigenKRAS Q61Honcogenic driver gene
spellingShingle Volker Lennerz
Volker Lennerz
Volker Lennerz
Christoph Doppler
Martina Fatho
Anja Dröge
Sigrid Schaper
Kristin Gennermann
Nadine Genzel
Stephanie Plassmann
David Weismann
Samuel W. Lukowski
Dominik Bents
Christina Beushausen
Karen Kriese
Hermann Herbst
Volkhard Seitz
Rudolf Hammer
Rudolf Hammer
Paul J. Adam
Stephan Eggeling
Catherine Wölfel
Thomas Wölfel
Steffen Hennig
Steffen Hennig
T-cell receptors identified by a personalized antigen-agnostic screening approach target shared neoantigen KRAS Q61H
Frontiers in Immunology
T-cell receptor (TCR)
TCR-T cell
tumor-specific antigen
neoantigen
KRAS Q61H
oncogenic driver gene
title T-cell receptors identified by a personalized antigen-agnostic screening approach target shared neoantigen KRAS Q61H
title_full T-cell receptors identified by a personalized antigen-agnostic screening approach target shared neoantigen KRAS Q61H
title_fullStr T-cell receptors identified by a personalized antigen-agnostic screening approach target shared neoantigen KRAS Q61H
title_full_unstemmed T-cell receptors identified by a personalized antigen-agnostic screening approach target shared neoantigen KRAS Q61H
title_short T-cell receptors identified by a personalized antigen-agnostic screening approach target shared neoantigen KRAS Q61H
title_sort t cell receptors identified by a personalized antigen agnostic screening approach target shared neoantigen kras q61h
topic T-cell receptor (TCR)
TCR-T cell
tumor-specific antigen
neoantigen
KRAS Q61H
oncogenic driver gene
url https://www.frontiersin.org/articles/10.3389/fimmu.2025.1509855/full
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