Preclinical optimization of a GPC2-targeting CAR T-cell therapy for neuroblastoma
Background Although most patients with newly diagnosed high-risk neuroblastoma (NB) achieve remission after initial therapy, more than 50% experience late relapses caused by minimal residual disease (MRD) and succumb to their cancer. Therapeutic strategies to target MRD may benefit these children. W...
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BMJ Publishing Group
2023-01-01
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| Series: | Journal for ImmunoTherapy of Cancer |
| Online Access: | https://jitc.bmj.com/content/11/1/e005881.full |
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| author | Nan Li Haiying Qin Rosa Nguyen Mitchell Ho Ming Sun Eytan Ruppin Michael C Kelly Yingying Cao Reona Okada Jeyshka M Reyes-González Hannah G Stack Charlie Seibert Carol J Thiele |
| author_facet | Nan Li Haiying Qin Rosa Nguyen Mitchell Ho Ming Sun Eytan Ruppin Michael C Kelly Yingying Cao Reona Okada Jeyshka M Reyes-González Hannah G Stack Charlie Seibert Carol J Thiele |
| author_sort | Nan Li |
| collection | DOAJ |
| description | Background Although most patients with newly diagnosed high-risk neuroblastoma (NB) achieve remission after initial therapy, more than 50% experience late relapses caused by minimal residual disease (MRD) and succumb to their cancer. Therapeutic strategies to target MRD may benefit these children. We developed a new chimeric antigen receptor (CAR) targeting glypican-2 (GPC2) and conducted iterative preclinical engineering of the CAR structure to maximize its anti-tumor efficacy before clinical translation.Methods We evaluated different GPC2-CAR constructs by measuring the CAR activity in vitro. NOD-SCID mice engrafted orthotopically with human NB cell lines or patient-derived xenografts and treated with human CAR T cells served as in vivo models. Mechanistic studies were performed using single-cell RNA-sequencing.Results Applying stringent in vitro assays and orthotopic in vivo NB models, we demonstrated that our single-chain variable fragment, CT3, integrated into a CAR vector with a CD28 hinge, CD28 transmembrane, and 4-1BB co-stimulatory domain (CT3.28H.BBζ) elicits the best preclinical anti-NB activity compared with other tested CAR constructs. This enhanced activity was associated with an enrichment of CD8+ effector T cells in the tumor-microenvironment and upregulation of several effector molecules such as GNLY, GZMB, ZNF683, and HMGN2. Finally, we also showed that the CT3.28H.BBζ CAR we developed was more potent than a recently clinically tested GD2-targeted CAR to control NB growth in vivo.Conclusion Given the robust preclinical activity of CT3.28H.BBζ, these results form a promising basis for further clinical testing in children with NB. |
| format | Article |
| id | doaj-art-c2a4dbca004d4ad9aeecc39207e2e0f9 |
| institution | Kabale University |
| issn | 2051-1426 |
| language | English |
| publishDate | 2023-01-01 |
| publisher | BMJ Publishing Group |
| record_format | Article |
| series | Journal for ImmunoTherapy of Cancer |
| spelling | doaj-art-c2a4dbca004d4ad9aeecc39207e2e0f92025-01-29T10:35:09ZengBMJ Publishing GroupJournal for ImmunoTherapy of Cancer2051-14262023-01-0111110.1136/jitc-2022-005881Preclinical optimization of a GPC2-targeting CAR T-cell therapy for neuroblastomaNan Li0Haiying Qin1Rosa Nguyen2Mitchell Ho3Ming Sun4Eytan Ruppin5Michael C Kelly6Yingying Cao7Reona Okada8Jeyshka M Reyes-González9Hannah G Stack10Charlie Seibert11Carol J Thiele12Laboratory of Molecular Biology, National Institutes of Health, Bethesda, Maryland, USAPediatric Oncology Branch, NCI, Bethesda, Maryland, USAPediatric Oncology Branch, NCI, Bethesda, Maryland, USA2NIH, Bethesda, MD, USAPediatric Oncology Branch, NCI, Bethesda, Maryland, USA23 Cancer Data Science Laboratory, National Cancer Institute, Bethesda, Maryland, USACenter for Childhood Cancer and Blood Disorders, Abigail Wexner Research Institute at Nationwide Children`s Hospital, Columbus, Ohio, USACancer Data Science Laboratory, NCI, Bethesda, Maryland, USAPediatric Oncology Branch, NCI, Bethesda, Maryland, USAPediatric Oncology Branch, NCI, Bethesda, Maryland, USAPediatric Oncology Branch, NCI, Bethesda, Maryland, USACenter for Cancer Research Single Cell Analysis Facility, Frederick National Laboratory for Cancer Research, Frederick, Maryland, USAPediatric Oncology Branch, NCI, Bethesda, Maryland, USABackground Although most patients with newly diagnosed high-risk neuroblastoma (NB) achieve remission after initial therapy, more than 50% experience late relapses caused by minimal residual disease (MRD) and succumb to their cancer. Therapeutic strategies to target MRD may benefit these children. We developed a new chimeric antigen receptor (CAR) targeting glypican-2 (GPC2) and conducted iterative preclinical engineering of the CAR structure to maximize its anti-tumor efficacy before clinical translation.Methods We evaluated different GPC2-CAR constructs by measuring the CAR activity in vitro. NOD-SCID mice engrafted orthotopically with human NB cell lines or patient-derived xenografts and treated with human CAR T cells served as in vivo models. Mechanistic studies were performed using single-cell RNA-sequencing.Results Applying stringent in vitro assays and orthotopic in vivo NB models, we demonstrated that our single-chain variable fragment, CT3, integrated into a CAR vector with a CD28 hinge, CD28 transmembrane, and 4-1BB co-stimulatory domain (CT3.28H.BBζ) elicits the best preclinical anti-NB activity compared with other tested CAR constructs. This enhanced activity was associated with an enrichment of CD8+ effector T cells in the tumor-microenvironment and upregulation of several effector molecules such as GNLY, GZMB, ZNF683, and HMGN2. Finally, we also showed that the CT3.28H.BBζ CAR we developed was more potent than a recently clinically tested GD2-targeted CAR to control NB growth in vivo.Conclusion Given the robust preclinical activity of CT3.28H.BBζ, these results form a promising basis for further clinical testing in children with NB.https://jitc.bmj.com/content/11/1/e005881.full |
| spellingShingle | Nan Li Haiying Qin Rosa Nguyen Mitchell Ho Ming Sun Eytan Ruppin Michael C Kelly Yingying Cao Reona Okada Jeyshka M Reyes-González Hannah G Stack Charlie Seibert Carol J Thiele Preclinical optimization of a GPC2-targeting CAR T-cell therapy for neuroblastoma Journal for ImmunoTherapy of Cancer |
| title | Preclinical optimization of a GPC2-targeting CAR T-cell therapy for neuroblastoma |
| title_full | Preclinical optimization of a GPC2-targeting CAR T-cell therapy for neuroblastoma |
| title_fullStr | Preclinical optimization of a GPC2-targeting CAR T-cell therapy for neuroblastoma |
| title_full_unstemmed | Preclinical optimization of a GPC2-targeting CAR T-cell therapy for neuroblastoma |
| title_short | Preclinical optimization of a GPC2-targeting CAR T-cell therapy for neuroblastoma |
| title_sort | preclinical optimization of a gpc2 targeting car t cell therapy for neuroblastoma |
| url | https://jitc.bmj.com/content/11/1/e005881.full |
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