Tumor-derived G-CSF induces an immunosuppressive microenvironment in an osteosarcoma model, reducing response to CAR.GD2 T-cells
Abstract Sarcomas are rare, mesenchymal tumors, representing about 10–15% of all childhood cancers. GD2 is a suitable target for chimeric antigen receptor (CAR) T-cell therapy due to its overexpression in several solid tumors. In this preclinical study, we investigated the potential use of iCasp9.2A...
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2024-12-01
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| Series: | Journal of Hematology & Oncology |
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| Online Access: | https://doi.org/10.1186/s13045-024-01641-7 |
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| author | Michele Pezzella Concetta Quintarelli Maria C. Quadraccia Andrea Sarcinelli Simona Manni Laura Iaffaldano Alessio Ottaviani Roselia Ciccone Antonio Camera Maria L. D’Amore Stefano Di Cecca Matilde Sinibaldi Marika Guercio Mariasole Aurigemma Pamela De Falco Valentina Fustaino Rossella Rota Silvia Pomella Matteo Cassandri Angela Di Giannatale Chiara Agrati Veronica Bordoni Federica Guarracino Michele Massa Giada Del Baldo Marco Becilli Giuseppe M. Milano Francesca Del Bufalo Franco Locatelli Biagio De Angelis |
| author_facet | Michele Pezzella Concetta Quintarelli Maria C. Quadraccia Andrea Sarcinelli Simona Manni Laura Iaffaldano Alessio Ottaviani Roselia Ciccone Antonio Camera Maria L. D’Amore Stefano Di Cecca Matilde Sinibaldi Marika Guercio Mariasole Aurigemma Pamela De Falco Valentina Fustaino Rossella Rota Silvia Pomella Matteo Cassandri Angela Di Giannatale Chiara Agrati Veronica Bordoni Federica Guarracino Michele Massa Giada Del Baldo Marco Becilli Giuseppe M. Milano Francesca Del Bufalo Franco Locatelli Biagio De Angelis |
| author_sort | Michele Pezzella |
| collection | DOAJ |
| description | Abstract Sarcomas are rare, mesenchymal tumors, representing about 10–15% of all childhood cancers. GD2 is a suitable target for chimeric antigen receptor (CAR) T-cell therapy due to its overexpression in several solid tumors. In this preclinical study, we investigated the potential use of iCasp9.2A.GD2.CAR-CD28.4–1BBζ (CAR.GD2) T-cells as a treatment option for patients who have GD2-positive sarcomas and we sought to identify factors shaping hostile tumor microenvironment in this setting. GD2 expression was evaluated by flow-cytometry on primary tumor biopsies of pediatric sarcoma patients. GD2 expression in sarcoma cells was also evaluated in response to an enhancer of zeste homolog 2 (EZH2) inhibitor (Tazemetostat). The antitumor activity of CAR.GD2 T-cells was evaluated both in vitro and in vivo preclinical models of orthotopic and/or metastatic soft-tissue and bone sarcomas. GD2 expression was detected in 55% of the primary tumors. Notably, the Osteosarcoma and Alveolar Rhabdomyosarcomas subtypes exhibited the highest GD2 expression levels, while Ewing sarcoma showed the lowest. CAR.GD2 T-cells show a significant tumor control both in vitro and in vivo models of GD2-expressing tumors. Pretreatment with an EZH2 inhibitor (Tazemetostat) upregulating GD2 expression, sensitizes GD2dim sarcoma cells to CAR.GD2 T-cells cytotoxic activity. Moreover, in mouse models of disseminated Rhabdomyosarcomas and orthotopic Osteosarcoma, CAR.GD2 T-cells showed both a vigorous anti-tumor activity and long-term persistence as compared to un-transduced T-cells. The presence of immunosuppressive murine myeloid-derived suppressor (MDSC) cells significantly reduces long-term anti-tumour activity of infused CAR.GD2 T-cells. Tumor-derived G-CSF was found to be one of the key factors driving expansion of immunosuppressive murine and human MDSC, thus indirectly limiting the efficacy of CAR.GD2 T-cells. Our preclinical data strongly suggest that CAR.GD2 T-cells hold promise as a potential therapeutic option for the treatment of patients with GD2-positive sarcomas. Strategies to tackle hostile immunosuppressive MDSC are desirable to optimize CAR.GD2 T-cell activity. |
| format | Article |
| id | doaj-art-d31ce40cc6e6413095e8c1ada8b07976 |
| institution | DOAJ |
| issn | 1756-8722 |
| language | English |
| publishDate | 2024-12-01 |
| publisher | BMC |
| record_format | Article |
| series | Journal of Hematology & Oncology |
| spelling | doaj-art-d31ce40cc6e6413095e8c1ada8b079762025-08-20T02:39:51ZengBMCJournal of Hematology & Oncology1756-87222024-12-0117112310.1186/s13045-024-01641-7Tumor-derived G-CSF induces an immunosuppressive microenvironment in an osteosarcoma model, reducing response to CAR.GD2 T-cellsMichele Pezzella0Concetta Quintarelli1Maria C. Quadraccia2Andrea Sarcinelli3Simona Manni4Laura Iaffaldano5Alessio Ottaviani6Roselia Ciccone7Antonio Camera8Maria L. D’Amore9Stefano Di Cecca10Matilde Sinibaldi11Marika Guercio12Mariasole Aurigemma13Pamela De Falco14Valentina Fustaino15Rossella Rota16Silvia Pomella17Matteo Cassandri18Angela Di Giannatale19Chiara Agrati20Veronica Bordoni21Federica Guarracino22Michele Massa23Giada Del Baldo24Marco Becilli25Giuseppe M. Milano26Francesca Del Bufalo27Franco Locatelli28Biagio De Angelis29Department of Onco-Haematology and Cell and Gene Therapy, Bambino Gesù Children’s Hospital, IRCCSDepartment of Onco-Haematology and Cell and Gene Therapy, Bambino Gesù Children’s Hospital, IRCCSDepartment of Onco-Haematology and Cell and Gene Therapy, Bambino Gesù Children’s Hospital, IRCCSDepartment of Onco-Haematology and Cell and Gene Therapy, Bambino Gesù Children’s Hospital, IRCCSDepartment of Onco-Haematology and Cell and Gene Therapy, Bambino Gesù Children’s Hospital, IRCCSDepartment of Onco-Haematology and Cell and Gene Therapy, Bambino Gesù Children’s Hospital, IRCCSDepartment of Onco-Haematology and Cell and Gene Therapy, Bambino Gesù Children’s Hospital, IRCCSDepartment of Onco-Haematology and Cell and Gene Therapy, Bambino Gesù Children’s Hospital, IRCCSDepartment of Onco-Haematology and Cell and Gene Therapy, Bambino Gesù Children’s Hospital, IRCCSDepartment of Onco-Haematology and Cell and Gene Therapy, Bambino Gesù Children’s Hospital, IRCCSDepartment of Onco-Haematology and Cell and Gene Therapy, Bambino Gesù Children’s Hospital, IRCCSDepartment of Onco-Haematology and Cell and Gene Therapy, Bambino Gesù Children’s Hospital, IRCCSDepartment of Onco-Haematology and Cell and Gene Therapy, Bambino Gesù Children’s Hospital, IRCCSDepartment of Onco-Haematology and Cell and Gene Therapy, Bambino Gesù Children’s Hospital, IRCCSDepartment of Onco-Haematology and Cell and Gene Therapy, Bambino Gesù Children’s Hospital, IRCCSDepartment of Onco-Haematology and Cell and Gene Therapy, Bambino Gesù Children’s Hospital, IRCCSDepartment of Onco-Haematology and Cell and Gene Therapy, Bambino Gesù Children’s Hospital, IRCCSDepartment of Onco-Haematology and Cell and Gene Therapy, Bambino Gesù Children’s Hospital, IRCCSDepartment of Onco-Haematology and Cell and Gene Therapy, Bambino Gesù Children’s Hospital, IRCCSDepartment of Onco-Haematology and Cell and Gene Therapy, Bambino Gesù Children’s Hospital, IRCCSUnit of Pathogen Specific Immunity, Bambino Gesù Children’s Hospital, IRCCSUnit of Pathogen Specific Immunity, Bambino Gesù Children’s Hospital, IRCCSUnit of Pathogen Specific Immunity, Bambino Gesù Children’s Hospital, IRCCSDepartment of Onco-Haematology and Cell and Gene Therapy, Bambino Gesù Children’s Hospital, IRCCSDepartment of Onco-Haematology and Cell and Gene Therapy, Bambino Gesù Children’s Hospital, IRCCSDepartment of Onco-Haematology and Cell and Gene Therapy, Bambino Gesù Children’s Hospital, IRCCSDepartment of Onco-Haematology and Cell and Gene Therapy, Bambino Gesù Children’s Hospital, IRCCSDepartment of Onco-Haematology and Cell and Gene Therapy, Bambino Gesù Children’s Hospital, IRCCSDepartment of Onco-Haematology and Cell and Gene Therapy, Bambino Gesù Children’s Hospital, IRCCSDepartment of Onco-Haematology and Cell and Gene Therapy, Bambino Gesù Children’s Hospital, IRCCSAbstract Sarcomas are rare, mesenchymal tumors, representing about 10–15% of all childhood cancers. GD2 is a suitable target for chimeric antigen receptor (CAR) T-cell therapy due to its overexpression in several solid tumors. In this preclinical study, we investigated the potential use of iCasp9.2A.GD2.CAR-CD28.4–1BBζ (CAR.GD2) T-cells as a treatment option for patients who have GD2-positive sarcomas and we sought to identify factors shaping hostile tumor microenvironment in this setting. GD2 expression was evaluated by flow-cytometry on primary tumor biopsies of pediatric sarcoma patients. GD2 expression in sarcoma cells was also evaluated in response to an enhancer of zeste homolog 2 (EZH2) inhibitor (Tazemetostat). The antitumor activity of CAR.GD2 T-cells was evaluated both in vitro and in vivo preclinical models of orthotopic and/or metastatic soft-tissue and bone sarcomas. GD2 expression was detected in 55% of the primary tumors. Notably, the Osteosarcoma and Alveolar Rhabdomyosarcomas subtypes exhibited the highest GD2 expression levels, while Ewing sarcoma showed the lowest. CAR.GD2 T-cells show a significant tumor control both in vitro and in vivo models of GD2-expressing tumors. Pretreatment with an EZH2 inhibitor (Tazemetostat) upregulating GD2 expression, sensitizes GD2dim sarcoma cells to CAR.GD2 T-cells cytotoxic activity. Moreover, in mouse models of disseminated Rhabdomyosarcomas and orthotopic Osteosarcoma, CAR.GD2 T-cells showed both a vigorous anti-tumor activity and long-term persistence as compared to un-transduced T-cells. The presence of immunosuppressive murine myeloid-derived suppressor (MDSC) cells significantly reduces long-term anti-tumour activity of infused CAR.GD2 T-cells. Tumor-derived G-CSF was found to be one of the key factors driving expansion of immunosuppressive murine and human MDSC, thus indirectly limiting the efficacy of CAR.GD2 T-cells. Our preclinical data strongly suggest that CAR.GD2 T-cells hold promise as a potential therapeutic option for the treatment of patients with GD2-positive sarcomas. Strategies to tackle hostile immunosuppressive MDSC are desirable to optimize CAR.GD2 T-cell activity.https://doi.org/10.1186/s13045-024-01641-7GD2Chimeric antigen receptor (CAR)SarcomaICasp9.2A.GD2.CAR-CD28.4–1BBζCAR.GD2 T-cellsEZH2 inhibitor |
| spellingShingle | Michele Pezzella Concetta Quintarelli Maria C. Quadraccia Andrea Sarcinelli Simona Manni Laura Iaffaldano Alessio Ottaviani Roselia Ciccone Antonio Camera Maria L. D’Amore Stefano Di Cecca Matilde Sinibaldi Marika Guercio Mariasole Aurigemma Pamela De Falco Valentina Fustaino Rossella Rota Silvia Pomella Matteo Cassandri Angela Di Giannatale Chiara Agrati Veronica Bordoni Federica Guarracino Michele Massa Giada Del Baldo Marco Becilli Giuseppe M. Milano Francesca Del Bufalo Franco Locatelli Biagio De Angelis Tumor-derived G-CSF induces an immunosuppressive microenvironment in an osteosarcoma model, reducing response to CAR.GD2 T-cells Journal of Hematology & Oncology GD2 Chimeric antigen receptor (CAR) Sarcoma ICasp9.2A.GD2.CAR-CD28.4–1BBζ CAR.GD2 T-cells EZH2 inhibitor |
| title | Tumor-derived G-CSF induces an immunosuppressive microenvironment in an osteosarcoma model, reducing response to CAR.GD2 T-cells |
| title_full | Tumor-derived G-CSF induces an immunosuppressive microenvironment in an osteosarcoma model, reducing response to CAR.GD2 T-cells |
| title_fullStr | Tumor-derived G-CSF induces an immunosuppressive microenvironment in an osteosarcoma model, reducing response to CAR.GD2 T-cells |
| title_full_unstemmed | Tumor-derived G-CSF induces an immunosuppressive microenvironment in an osteosarcoma model, reducing response to CAR.GD2 T-cells |
| title_short | Tumor-derived G-CSF induces an immunosuppressive microenvironment in an osteosarcoma model, reducing response to CAR.GD2 T-cells |
| title_sort | tumor derived g csf induces an immunosuppressive microenvironment in an osteosarcoma model reducing response to car gd2 t cells |
| topic | GD2 Chimeric antigen receptor (CAR) Sarcoma ICasp9.2A.GD2.CAR-CD28.4–1BBζ CAR.GD2 T-cells EZH2 inhibitor |
| url | https://doi.org/10.1186/s13045-024-01641-7 |
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