{"doi":"10.1136/jitc-2024-010239","title":"TIM-3 blockade enhances ex vivo stimulated allogeneic NK cell therapy for relapsed murine neuroblastoma after hematopoietic cell transplant","abstract":"BACKGROUND: High-risk neuroblastoma (HR-NBL) is an aggressive tumor of the sympathetic nervous system with high risk of relapse and poor overall survival. Allogeneic hematopoietic cell transplant (allo-HCT) has been used previously in patients with HR-NBL; however, graft-versus-host disease (GVHD) and disease progression have limited clinical application. Ex vivo stimulated allogeneic natural killer (NK) cells represent an approach to enhance the graft-versus-tumor (GVT) effect without exacerbation of GVHD but have not shown efficacy in NBL. METHODS: aggressive variant of the Neuro-2a murine neuroblastoma cell line (15-4P) at a 1:1 ratio for 10-12 days. Allogeneic NK cells were then analyzed for activation, proliferation, cytokine production, and cytotoxicity against two murine NBL cell lines, Neuro2a and NXS2, in the absence or presence of anti-T-cell immunoglobulin and mucin-domain containing-3 (TIM-3). Lethally irradiated B6AJF1 mice received allo-HCT from B6 donors followed by NBL challenge after 7 days to mimic tumor relapse. Select groups received anti-TIM-3 starting on day 9 for every 4 days with/without infusions of 15-4P B6 NK cells on days 14, 21, and 28. In select experiments, T cell and NK cells were selectively depleted to establish contribution to the GVT effect. All groups were analyzed for tumor growth, GVHD and survival. RESULTS: Co-culturing NK cells with 15-4P results in 78-fold expansion with increased expression of Kiel-67 (Ki-67) and Natural Killer Group 2, Member D (NKG2D), NKp46, TNF-Related Apoptosis-Inducing Ligand (TRAIL) and TIM-3. 15-4P stimulated allogeneic NK cells showed enhanced cytotoxicity against NBL compared with IL-15 NK cells alone but was limited in part due to high expression of TIM-3 ligands on Neuro-2a compared with NXS2. The addition of TIM-3 blockade further enhanced NK cytotoxicity versus Neuro-2a, with enhanced 15-4P NK cell degranulation, Eomesodermin, TRAIL and Fas Ligand expression observed. In vivo, the combination of 15-4P stimulated allogeneic NK cells and TIM-3 blockade after allo-HCT resulted in prolonged survival against NBL with decreased tumor burden compared with NK cells or anti-TIM-3 alone. Depletion of NK cells, but not T cells, abrogated the GVT effect. CONCLUSION: Allo-HCT can be a platform for treating NBL using combination ex vivo stimulated allogeneic NK cell therapy with TIM-3 blockade to enhance the GVT effect without inducing GVHD.","journal":"Journal for ImmunoTherapy of Cancer","year":2025,"id":549382,"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":1,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9599,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2025-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":1296506,"name":"Evan L. Dray","orcid":"0000-0002-9293-7583","position":1,"is_corresponding":false},{"id":998762,"name":"Nicholas R Mohrdieck","orcid":null,"position":2,"is_corresponding":false},{"id":998763,"name":"Chloe King","orcid":null,"position":3,"is_corresponding":false},{"id":998764,"name":"Anastasia A Griggs","orcid":null,"position":4,"is_corresponding":false},{"id":1169583,"name":"Jillian M Kline","orcid":null,"position":5,"is_corresponding":false},{"id":419903,"name":"Monica M. Cho","orcid":"0000-0002-6708-9578","position":6,"is_corresponding":false},{"id":606012,"name":"Sean Rinella","orcid":null,"position":7,"is_corresponding":false},{"id":998765,"name":"Katharine E. Tippins","orcid":null,"position":8,"is_corresponding":false},{"id":738569,"name":"Paul D. Bates","orcid":"0009-0001-2418-0312","position":9,"is_corresponding":false},{"id":258238,"name":"Lei Shi","orcid":"0000-0002-5312-8521","position":10,"is_corresponding":false},{"id":1169582,"name":"Longzhen Song","orcid":null,"position":11,"is_corresponding":false},{"id":338812,"name":"Nicholas J Hess","orcid":"0000-0001-5720-9305","position":12,"is_corresponding":false},{"id":998185,"name":"Tyce Kearl","orcid":"0000-0002-9255-892X","position":13,"is_corresponding":false},{"id":658163,"name":"Bryon D. Johnson","orcid":"0000-0001-7416-3396","position":14,"is_corresponding":false},{"id":277448,"name":"Christian M. Capitini","orcid":"0000-0002-2276-6731","position":15,"is_corresponding":false},{"id":419904,"name":"Aicha Quamine","orcid":"0000-0002-9049-7078","position":0,"is_corresponding":true}],"reference_count":46,"raw_metadata":null,"created_at":"2026-07-19T02:54:07.823422Z","pmid":"41386972","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":[]}