{"doi":"10.1136/jitc-2022-005881","title":"Preclinical optimization of a GPC2-targeting CAR T-cell therapy for neuroblastoma","abstract":"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.","journal":"Journal for ImmunoTherapy of Cancer","year":2023,"id":324892,"datarank":0.0,"base_score":0.0,"endowment":0.0,"self_citation_contribution":0.0,"citation_network_contribution":0.0,"self_endowment_contribution":0.0,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":32,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9501,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2023-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":991685,"name":"Yingying Cao","orcid":"0000-0003-2812-2299","position":1,"is_corresponding":false},{"id":1036274,"name":"Reona Okada","orcid":"0000-0002-2584-3026","position":2,"is_corresponding":false},{"id":499935,"name":"Jeyshka M. Reyes‐González","orcid":null,"position":3,"is_corresponding":false},{"id":1041726,"name":"Hannah G Stack","orcid":"0009-0005-5629-601X","position":4,"is_corresponding":false},{"id":309021,"name":"Haiying Qin","orcid":"0000-0001-5966-8812","position":5,"is_corresponding":false},{"id":255115,"name":"Nan Li","orcid":"0000-0002-1498-7139","position":6,"is_corresponding":false},{"id":564135,"name":"Charlie Seibert","orcid":null,"position":7,"is_corresponding":false},{"id":226280,"name":"Michael C. Kelly","orcid":"0000-0003-0654-2778","position":8,"is_corresponding":false},{"id":440002,"name":"Fiorella Schischlik","orcid":"0000-0003-4299-7657","position":9,"is_corresponding":false},{"id":255131,"name":"Mitchell Ho","orcid":"0000-0002-9152-5405","position":10,"is_corresponding":false},{"id":399507,"name":"Carol J. Thiele","orcid":"0000-0003-2869-7690","position":11,"is_corresponding":false},{"id":267014,"name":"Rosa Nguyen","orcid":"0000-0001-5143-2055","position":12,"is_corresponding":false},{"id":563031,"name":"Ming Sun","orcid":"0000-0002-6313-4458","position":0,"is_corresponding":true}],"reference_count":43,"raw_metadata":null,"created_at":"2026-07-19T01:08:16.809511Z","pmid":"36631162","pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}