Christina Dargel1, Michal Bassani-Sternberg2, Julia Hasreiter1, Fabio Zani3, Jan-Hendrik Bockmann4, Frank Thiele4, Felix Bohne1, Karin Wisskirchen1, Susanne Wilde5, Martin F Sprinzl6, Dolores J Schendel7, Angela M Krackhardt8, Wolfgang Uckert9, Dirk Wohlleber10, Matthias Schiemann11, Kerstin Stemmer3, Mathias Heikenwälder1, Dirk H Busch12, Günther Richter13, Matthias Mann2, Ulrike Protzer14. 1. Institute of Virology, Technische Universität München, Helmholtz Zentrum München, München, Germany. 2. Max Planck Institute of Biochemistry, Martinsried, Germany. 3. Institute for Diabetes and Obesity, Helmholtz Zentrum München, Garching, Germany. 4. Institute of Virology, Technische Universität München, Helmholtz Zentrum München, München, Germany; German Center for Infection Research (DZIF), Munich Site, Germany. 5. Institute of Molecular Immunology, Helmholtz Zentrum München, München, Germany. 6. I. Medizinische Klinik und Poliklinik, Universitätsmedizin der Johannes Gutenberg-Universität, Mainz, Germany. 7. Institute of Molecular Immunology, Helmholtz Zentrum München, München, Germany; Clinical Cooperation Groups Antigen Specific Immunotherapy and Immune Monitoring, Technische Universität München, Helmholtz Zentrum München, München, Germany. 8. Clinical Cooperation Groups Antigen Specific Immunotherapy and Immune Monitoring, Technische Universität München, Helmholtz Zentrum München, München, Germany; 3rd Medical Department, University Hospital Rechts der Isar, Technische Universität München, München, Germany. 9. Max-Delbrück-Centrum for Molecular Medicine (MDC) and Institute of Biology, Humboldt University Berlin, Berlin-Buch, Germany. 10. Institute of Molecular Immunology, University Hospital Rechts der Isar, Technische Universität München, München, Germany. 11. Clinical Cooperation Groups Antigen Specific Immunotherapy and Immune Monitoring, Technische Universität München, Helmholtz Zentrum München, München, Germany; Institute for Medical Microbiology, Immunology and Hygiene, Technische Universität München, München, Germany. 12. German Center for Infection Research (DZIF), Munich Site, Germany; Clinical Cooperation Groups Antigen Specific Immunotherapy and Immune Monitoring, Technische Universität München, Helmholtz Zentrum München, München, Germany; Institute for Medical Microbiology, Immunology and Hygiene, Technische Universität München, München, Germany. 13. Department of Pediatrics, University Hospital Rechts der Isar, Technische Universität München, München, Germany. 14. Institute of Virology, Technische Universität München, Helmholtz Zentrum München, München, Germany; German Center for Infection Research (DZIF), Munich Site, Germany; Clinical Cooperation Groups Antigen Specific Immunotherapy and Immune Monitoring, Technische Universität München, Helmholtz Zentrum München, München, Germany. Electronic address: protzer@tum.de.
Abstract
BACKGROUND & AIMS: Cancer therapies are being developed based on our ability to direct T cells against tumor antigens. Glypican-3 (GPC3) is expressed by 75% of all hepatocellular carcinomas (HCC), but not in healthy liver tissue or other organs. We aimed to generate T cells with GPC3-specific receptors that recognize HCC and used them to eliminate GPC3-expressing xenograft tumors grown from human HCC cells in mice. METHODS: We used mass spectrometry to obtain a comprehensive peptidome from GPC3-expressing hepatoma cells after immune-affinity purification of human leukocyte antigen (HLA)-A2 and bioinformatics to identify immunodominant peptides. To circumvent GPC3 tolerance resulting from fetal expression, dendritic cells from HLA-A2-negative donors were cotransfected with GPC3 and HLA-A2 RNA to stimulate and expand antigen-specific T cells. RESULTS: Peptide GPC3367 was identified as a predominant peptide on HLA-A2. We used A2-GPC3367 multimers to detect, select for, and clone GPC3-specific T cells. These clones bound the A2-GPC3367 multimer and secreted interferon-γ when cultured with GPC3367, but not with control peptide-loaded cells. By genomic sequencing of these T-cell clones, we identified a gene encoding a dominant T-cell receptor. The gene was cloned and the sequence was codon optimized and expressed from a retroviral vector. Primary CD8(+) T cells that expressed the transgenic T-cell receptor specifically bound GPC3367 on HLA-A2. These T cells killed GPC3-expressing hepatoma cells in culture and slowed growth of HCC xenograft tumors in mice. CONCLUSIONS: We identified a GPC3367-specific T-cell receptor. Expression of this receptor by T cells allows them to recognize and kill GPC3-positive hepatoma cells. This finding could be used to advance development of adoptive T-cell therapy for HCC.
BACKGROUND & AIMS: Cancer therapies are being developed based on our ability to direct T cells against tumor antigens. Glypican-3 (GPC3) is expressed by 75% of all hepatocellular carcinomas (HCC), but not in healthy liver tissue or other organs. We aimed to generate T cells with GPC3-specific receptors that recognize HCC and used them to eliminate GPC3-expressing xenograft tumors grown from human HCC cells in mice. METHODS: We used mass spectrometry to obtain a comprehensive peptidome from GPC3-expressing hepatoma cells after immune-affinity purification of human leukocyte antigen (HLA)-A2 and bioinformatics to identify immunodominant peptides. To circumvent GPC3 tolerance resulting from fetal expression, dendritic cells from HLA-A2-negative donors were cotransfected with GPC3 and HLA-A2 RNA to stimulate and expand antigen-specific T cells. RESULTS: Peptide GPC3367 was identified as a predominant peptide on HLA-A2. We used A2-GPC3367 multimers to detect, select for, and clone GPC3-specific T cells. These clones bound the A2-GPC3367 multimer and secreted interferon-γ when cultured with GPC3367, but not with control peptide-loaded cells. By genomic sequencing of these T-cell clones, we identified a gene encoding a dominant T-cell receptor. The gene was cloned and the sequence was codon optimized and expressed from a retroviral vector. Primary CD8(+) T cells that expressed the transgenic T-cell receptor specifically bound GPC3367 on HLA-A2. These T cells killed GPC3-expressing hepatoma cells in culture and slowed growth of HCC xenograft tumors in mice. CONCLUSIONS: We identified a GPC3367-specific T-cell receptor. Expression of this receptor by T cells allows them to recognize and kill GPC3-positive hepatoma cells. This finding could be used to advance development of adoptive T-cell therapy for HCC.
Authors: David Schirmer; Thomas G P Grünewald; Richard Klar; Oxana Schmidt; Dirk Wohlleber; Rebeca Alba Rubío; Wolfgang Uckert; Uwe Thiel; Felix Bohne; Dirk H Busch; Angela M Krackhardt; Stefan Burdach; Günther H S Richter Journal: Oncoimmunology Date: 2016-04-25 Impact factor: 8.110
Authors: Timothy T Spear; Glenda G Callender; Jeffrey J Roszkowski; Kelly M Moxley; Patricia E Simms; Kendra C Foley; David C Murray; Gina M Scurti; Mingli Li; Justin T Thomas; Alexander Langerman; Elizabeth Garrett-Mayer; Yi Zhang; Michael I Nishimura Journal: Cancer Immunol Immunother Date: 2016-02-03 Impact factor: 6.968
Authors: Solomon Owusu Sekyere; Bernhard Schlevogt; Friederike Mettke; Mohammad Kabbani; Katja Deterding; Thomas Christian Wirth; Arndt Vogel; Michael Peter Manns; Christine Susanne Falk; Markus Cornberg; Heiner Wedemeyer Journal: Liver Cancer Date: 2018-07-18 Impact factor: 11.740