Human HLA-A*02:01/CHM1+ allo-restricted T cell receptor transgenic CD8+ T cells specifically inhibit Ewing sarcoma growth in vitro and in vivo.
The endochondral bone protein Chondromodulin-I (CHM1) provides oncogene addiction in Ewing sarcoma (ES). We pre-clinically tested the targetability of CHM1 by TCR transgenic, allo-restricted, peptide specific T cells to treat ES. We previously generated allo-restricted wildtype CD8+ T cells directed against the ES specific antigen CHM1319 causing specific responses against ES. However, utilization of these cells in current therapy protocols is hampered due to high complexity in production, relatively low cell numbers, and rapid T cell exhaustion.In order to provide off-the-shelf products in the future, we successfully generated HLA-A*02:01-restricted T cell receptor (TCR) transgenic T cells directed against CHM1319 by retroviral transduction.After short-term expansion a 100% purified CHM1319-TCR-transgenic T cell population expressed a CD62L+/CD45RO and CD62L+/CD45RA+ phenotype. These cells displayed specific in vitro IFNg and granzyme B release in co-culture with HLA-A*02:01+ ES cell lines expressing CHM1. When co-injected with ES cells in Rag2-/-É£c-/- mice, CHM1-specific TCR-transgenic T cells significantly inhibited the formation of lung and liver metastases in contrast to control mice. Lungs and livers of representative mice displayed CD8+ T cell infiltration in the presence (control group treated with unspecific T cells) and in the absence (study group) of metastatic disease, respectively. Furthermore, mice receiving unspecific T cells showed signs of graft-versus-host-disease in contrast to all mice, receiving CHM1319-TCR-transgenic T cells.CHM1319 specific TCR-transgenic T cells were successfully generated causing anti-ES responses in vitro and in vivo. In the future, CHM1319-TCR-transgenic T cells may control minimal residual disease rendering donor lymphocyte infusions more efficacious and less toxic.
Laboratory for Functional Genomics and Transplantation Biology, Department of Pediatrics and Children's Cancer Research Center, Klinikum rechts der Isar, Technische Universitaet Muenchen, Munich, Germany.|Present address: Laboratory for Immunotherapy, Dr. von Hauner Children's Hospital, LMU München, Munich, Germany.
Franziska
F
Blaeschke
Laboratory for Functional Genomics and Transplantation Biology, Department of Pediatrics and Children's Cancer Research Center, Klinikum rechts der Isar, Technische Universitaet Muenchen, Munich, Germany.
Uwe
U
Thiel
Laboratory for Functional Genomics and Transplantation Biology, Department of Pediatrics and Children's Cancer Research Center, Klinikum rechts der Isar, Technische Universitaet Muenchen, Munich, Germany.
Andreas
A
Kirschner
Laboratory for Functional Genomics and Transplantation Biology, Department of Pediatrics and Children's Cancer Research Center, Klinikum rechts der Isar, Technische Universitaet Muenchen, Munich, Germany.
Melanie
M
Thiede
Laboratory for Pediatric Sarcoma Biology, Institute of Pathology of the LMU Munich, Munich, Germany.
Rebeca Alba
RA
Rubio
Laboratory for Functional Genomics and Transplantation Biology, Department of Pediatrics and Children's Cancer Research Center, Klinikum rechts der Isar, Technische Universitaet Muenchen, Munich, Germany.
David
D
Schirmer
Laboratory for Pediatric Sarcoma Biology, Institute of Pathology of the LMU Munich, Munich, Germany.|German Cancer Consortium (DKTK), Heidelberg, Germany.|German Cancer Research Center, Heidelberg, Germany.
Thomas
T
Kirchner
Laboratory for Functional Genomics and Transplantation Biology, Department of Pediatrics and Children's Cancer Research Center, Klinikum rechts der Isar, Technische Universitaet Muenchen, Munich, Germany.
Günther H S
Richter
Medizinische Klinik III, Klinikum rechts der Isar, Technische Universität München, Munich, Germany.
Sabine
S
Mall
Medizinische Klinik III, Klinikum rechts der Isar, Technische Universität München, Munich, Germany.
Richard
R
Klar
Department of Medicine, University of Washington, Seattle, WA, USA.
Stanley
S
Riddell
Institute for Medical Microbiology, Immunology and Hygiene, Technische Universitaet Muenchen, München, Germany.|Focus Group "Clinical Cell Processing and Purification", Institute for Advanced Study, Technische Universität München, Munich, Germany.
Dirk H
DH
Busch
Medizinische Klinik III, Klinikum rechts der Isar, Technische Universität München, Munich, Germany.
Angela
A
Krackhardt
Laboratory for Pediatric Sarcoma Biology, Institute of Pathology of the LMU Munich, Munich, Germany.|German Cancer Consortium (DKTK), Heidelberg, Germany.|German Cancer Research Center, Heidelberg, Germany.
Thomas G P
TG
Grunewald
Laboratory for Functional Genomics and Transplantation Biology, Department of Pediatrics and Children's Cancer Research Center, Klinikum rechts der Isar, Technische Universitaet Muenchen, Munich, Germany.|Munich Comprehensive Cancer Center (CCC), Klinikum rechts der Isar, Technische Universität München, Germany.
Stefan
S
Burdach
D000818:Animals; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology / Q000378:metabolism / Q000637:transplantation*; D045744:Cell Line, Tumor; D018920:Coculture Techniques; D004268:DNA-Binding Proteins / Q000235:genetics; D006086:Graft vs Host Disease / Q000276:immunology; D053804:Granzymes / Q000378:metabolism; D015234:HLA-A Antigens / Q000235:genetics / Q000276:immunology; D018380:Hematopoietic Stem Cell Transplantation / Q000009:adverse effects / Q000379:methods*; D006801:Humans; D016219:Immunotherapy, Adoptive / Q000009:adverse effects / Q000379:methods*; D036341:Intercellular Signaling Peptides and Proteins / Q000378:metabolism*; D007371:Interferon-gamma / Q000378:metabolism; D008099:Liver / Q000473:pathology; D008168:Lung / Q000473:pathology; D008565:Membrane Proteins / Q000378:metabolism*; D051379:Mice; D008807:Mice, Inbred BALB C; D000074723:Oncogene Addiction; D011948:Receptors, Antigen, T-Cell / Q000235:genetics; D012190:Retroviridae / Q000235:genetics; D012512:Sarcoma, Ewing / Q000276:immunology / Q000473:pathology / Q000628:therapy*; D014161:Transduction, Genetic; D014184:Transplantation, Homologous / Q000009:adverse effects / Q000379:methods
D004268:DNA-Binding Proteins; D015234:HLA-A Antigens; D036341:Intercellular Signaling Peptides and Proteins; D008565:Membrane Proteins; C499308:Rag2 protein, mouse; D011948:Receptors, Antigen, T-Cell; C068675:CNMD protein, human; D007371:Interferon-gamma; C509213:GZMB protein, human; D053804:Granzymes
Ewing sarcoma; T cell receptor transgenic T cells; adoptive transfer; allogeneic stem cell transplantation; immunotherapy