{"doi":"10.1136/jitc-2024-010618","title":"Differential impact of TIM-3 ligands on NK cell function","abstract":"Background The transmembrane protein T-cell immunoglobulin and mucin-domain containing molecule 3 (TIM-3) is an immune checkpoint receptor that is expressed by a variety of leukocyte subsets, particularly in the tumor microenvironment. An effective TIM-3-targeting therapy should account for multiple biological factors, including the disease setting, the specific cell types involved and their varying sensitivities to the four putative TIM-3 ligands (galectin-9, phosphatidylserine, high mobility group protein B1 and carcinoembryonic antigen cell adhesion molecule 1), each of which engages a unique binding site on the receptor’s variable immunoglobulin domain. The primary objectives of this study were to assess the prevalence and function of TIM-3 + natural killer (NK) cells in patients with head and neck squamous cell carcinoma (HNSCC), determine whether the four TIM-3 ligands differentially affect TIM-3 + NK cell functions, identify the most immunosuppressive ligand, and evaluate whether targeting ligand-mediated TIM-3 signaling enhances NK cell effector functions. Methods Single-cell RNA sequencing and flow cytometry were used to study the prevalence, phenotypes and function of TIM-3 + NK cells in HNSCC patient tumors and blood. In vitro killing, proliferation and cytokine production assays were implemented to evaluate whether the four TIM-3 ligands differentially modulate TIM-3 + NK cell functions, and whether disruption of TIM-3/ligand interaction can enhance NK cell-mediated antitumor effector mechanisms. Finally, The Cancer Genome Atlas survival analysis and digital spatial profiling were employed to study the potential impact of etiology-associated differences on patients with HNSCC outcomes. Results We demonstrate that TIM-3 is highly prevalent on circulating and tumor-infiltrating NK cells. It co-expresses with CD44 and marks NK cells with heightened effector potential. Among the four putative TIM-3 ligands, galectin-9 most consistently suppresses NK cell-mediated cytotoxicity and proliferation through TIM-3 and CD44 signaling, respectively, but promotes IFN-γ release in a TIM-3-dependent manner. Among patients with HNSCC, an elevated intratumoral TIM-3 + NK cell gene signature associates with worse outcomes, specifically in those with human papillomavirus (HPV) + disease, potentially attributable to higher galectin-9 levels in HPV + versus HPV − patients. Conclusions Our findings underscore the complex functional impact of TIM-3 ligand signaling, which is consistent with recent clinical trials suggesting that targeting TIM-3 alone is suboptimal as an immunotherapeutic approach for treating malignancies.","journal":"Journal for ImmunoTherapy of Cancer","year":2025,"id":510374,"datarank":0.4636563680037475,"base_score":3.091042453358316,"endowment":3.091042453358316,"self_citation_contribution":0.4636563680037475,"citation_network_contribution":0.0,"self_endowment_contribution":0.4636563680037475,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":21,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9603,"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":994120,"name":"Housaiyin Li","orcid":null,"position":1,"is_corresponding":false},{"id":449665,"name":"Aditi Kulkarni","orcid":"0000-0002-4886-6095","position":2,"is_corresponding":false},{"id":1268381,"name":"Jennifer L. Anderson","orcid":"0000-0002-1471-5803","position":3,"is_corresponding":false},{"id":1367454,"name":"Pragati Upadhyay","orcid":null,"position":4,"is_corresponding":false},{"id":1367455,"name":"Onyedikachi Victor Onyekachi","orcid":null,"position":5,"is_corresponding":false},{"id":965846,"name":"Lídia Maria Rebolho Batista Arantes","orcid":"0000-0001-8230-1218","position":6,"is_corresponding":false},{"id":638505,"name":"Hridesh Banerjee","orcid":"0000-0002-4536-4102","position":7,"is_corresponding":false},{"id":349057,"name":"Larry Kane","orcid":"0000-0001-5198-516X","position":8,"is_corresponding":false},{"id":1366683,"name":"Xin Zhang","orcid":"0000-0001-9392-5456","position":9,"is_corresponding":false},{"id":109276,"name":"Tullia C. Bruno","orcid":"0000-0002-6433-0207","position":10,"is_corresponding":false},{"id":35765,"name":"Riyue Bao","orcid":"0000-0002-6105-1704","position":11,"is_corresponding":false},{"id":109277,"name":"Robert L. Ferris","orcid":"0000-0001-6605-2071","position":12,"is_corresponding":false},{"id":619120,"name":"Lazar Vujanović","orcid":"0000-0003-1729-8619","position":13,"is_corresponding":false},{"id":1366682,"name":"Juncheng Wang","orcid":"0000-0001-9909-274X","position":0,"is_corresponding":true}],"reference_count":48,"raw_metadata":{"citation_network_status":"fetched"},"created_at":"2026-07-19T02:47:36.538276Z","pmid":"39773563","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":[]}