{"gene":"CLC","run_date":"2026-06-14T21:03:45+00:00","timeline":{"discoveries":[{"year":2002,"finding":"CLC protein (galectin-10) is NOT a lysophospholipase: antibody depletion of CLC from eosinophil lysates retained full lysophospholipase activity, and purified CLC protein lacked significant lysophospholipase activity. Instead, CLC protein binds lysophospholipase inhibitors (p-chloromercuribenzenesulfonate via disulfide bonds with Cys29 and Cys57, and N-ethylmaleimide via ring stacking with Trp72 in the carbohydrate recognition domain) and physically interacts with eosinophil lysophospholipases.","method":"Antibody affinity chromatography depletion, enzyme activity assays, X-ray crystallography of CLC–inhibitor complexes, ligand blotting","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1 / Strong — X-ray crystal structures plus enzymatic depletion assays plus ligand blotting; multiple orthogonal methods definitively establishing both a negative and a positive mechanistic finding in a single rigorous study","pmids":["11834744"],"is_preprint":false},{"year":1999,"finding":"CLC protein (galectin-10) selectively binds mannose (not β-galactosides) through its carbohydrate recognition domain (CRD); partial conservation of galectin CRD residues results in altered topology and chemistry of the binding site, explaining the switch in carbohydrate specificity.","method":"X-ray crystallography of CLC–mannose complex at 1.8 Å resolution","journal":"Biochemistry","confidence":"High","confidence_rationale":"Tier 1 / Strong — crystal structure with direct ligand complex at high resolution; single rigorous study with structural validation","pmids":["10529229"],"is_preprint":false},{"year":2007,"finding":"Galectin-10 is constitutively and exclusively expressed intracellularly in human CD4+CD25+Foxp3+ regulatory T cells (Tregs) but not in resting or activated conventional CD4+ T cells; specific inhibition of galectin-10 restored proliferative capacity of Tregs and abrogated their suppressive function, establishing galectin-10 as essential for Treg anergy and suppression.","method":"Differential proteomics, single-cell staining, flow cytometry, specific inhibition of galectin-10","journal":"Blood","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — proteomics identification confirmed at mRNA and protein levels, functional inhibition with defined phenotypic readout; single lab with multiple orthogonal methods","pmids":["17502455"],"is_preprint":false},{"year":2018,"finding":"Galectin-10 forms a novel homodimer with a global shape distinct from other prototype galectins (Gal-1, -2, -7); in the dimer, Glu33 from one subunit blocks the carbohydrate-binding site of the other subunit, inhibiting disaccharide binding. Glycerol and small hydroxylated molecules bind the ligand-binding site with His53 being most crucial. Trp72 negatively regulates ligand binding (W72A mutant shows enhanced erythrocyte agglutination).","method":"X-ray crystallography of Gal-10 and eight variants (1.55–2.00 Å resolution), size exclusion chromatography, hemagglutination assay, alanine mutagenesis","journal":"Glycobiology","confidence":"High","confidence_rationale":"Tier 1 / Strong — multiple crystal structures plus mutagenesis plus biochemical assays in one study; multiple orthogonal methods","pmids":["29293962"],"is_preprint":false},{"year":2019,"finding":"Glu33 in the galectin-10 dimer interface blocks lactose binding; E33A mutant adopts a conformation allowing lactose binding as shown structurally and biochemically. Trp127 at the homodimer interface is essential for dimerization; the W127A monomer shows higher hemagglutination activity and can bind lactose-modified sepharose, confirming that dimerization suppresses carbohydrate binding. Trp72 is required for nuclear transport of Gal-10 (EGFP-tagged W72A cannot be transported into the nucleus in HeLa cells).","method":"X-ray crystallography of E33A variant, hemagglutination assay, solid-phase binding assay, EGFP-tagged subcellular localization in HeLa cells, site-directed mutagenesis","journal":"Glycobiology","confidence":"High","confidence_rationale":"Tier 1 / Strong — structural plus biochemical plus cell imaging with mutagenesis; multiple orthogonal methods in one study","pmids":["30239701"],"is_preprint":false},{"year":2020,"finding":"CLC/Gal-10 interacts with eosinophil granule cationic ribonucleases (eosinophil-derived neurotoxin/RNS2 and eosinophil cationic protein/RNS3) and with murine eosinophil-associated RNases; this interaction is independent of glycosylation and is not inhibitory to endoRNase activity. Knockdown of CLC/Gal-10 in human CD34+ cord blood-derived progenitor eosinophils impairs eosinophil granulogenesis. IFN-γ activation induces rapid colocalization of CLC/Gal-10 with EDN/RNS2 and CD63, suggesting a role in vesicular transport of cationic RNases during degranulation.","method":"Ligand blotting, co-immunoprecipitation, co-affinity purification, enzyme activity assays, confocal microscopy, shRNA knockdown in CD34+ progenitor differentiation model","journal":"The Journal of allergy and clinical immunology","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal co-IP/affinity purification plus shRNA KD with defined granulogenesis phenotype plus confocal imaging; multiple orthogonal methods","pmids":["31982451"],"is_preprint":false},{"year":2020,"finding":"CLC-P/Gal-10 is stored predominantly in the peripheral cytoplasm of human eosinophils (within ~250 nm of the plasma membrane) and NOT within secretory (specific) granules; stimulation with CCL11 or TNF-α does not change this peripheral localization, indicating CLC-P/Gal-10 is not exported through classical degranulation (piecemeal degranulation or compound exocytosis). High-density microdomains of CLC-P/Gal-10 interact with the plasma membrane in ~60% of the membrane area.","method":"Pre-embedding immunonanogold transmission electron microscopy, quantitative imaging analysis, immunofluorescence, eosinophil activation with CCL11 and TNF-α","journal":"Journal of leukocyte biology","confidence":"High","confidence_rationale":"Tier 2 / Strong — high-resolution immuno-EM with quantitative analysis plus immunofluorescence; orthogonal localization methods with functional context","pmids":["32108369"],"is_preprint":false},{"year":2021,"finding":"EETosis (eosinophil extracellular trap cell death) is the mechanism for galectin-10 release from eosinophils; EETosis is dependent on reactive oxygen species and PAD4-dependent histone citrullination, resulting in cytolytic release of net-like extracellular traps containing galectin-10, free galectin-10, and membrane-bound intact granules. Loss of cytoplasmic galectin-10 and extracellular deposition are signatures of EETosis in eosinophilic granulomatosis with polyangiitis (EGPA) tissues.","method":"In vitro eosinophil EETosis characterization, ROS and PAD4 inhibition assays, immunostaining, electron microscopy of EGPA tissues, ELISA","journal":"Arthritis & rheumatology (Hoboken, N.J.)","confidence":"High","confidence_rationale":"Tier 2 / Strong — in vitro mechanistic dissection with inhibitors plus EM and immunostaining in patient tissues; multiple orthogonal methods","pmids":["33750029"],"is_preprint":false},{"year":2017,"finding":"Galectin-10 functions as a T cell-suppressive molecule in eosinophils: antibody-mediated neutralization of galectin-10 partially abrogated eosinophil-mediated suppression of T cell proliferation; recombinant galectin-10 alone suppressed T cell proliferation. Galectin-10-containing immune synapses form between eosinophils and lymphocytes.","method":"Antibody neutralization in coculture, recombinant protein functional assay, FACS sorting, immunofluorescence imaging of immune synapses","journal":"Journal of immunology (Baltimore, Md. : 1950)","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — neutralization plus recombinant protein functional assay plus imaging; single lab with multiple orthogonal methods","pmids":["28515279"],"is_preprint":false},{"year":2020,"finding":"CD16+ suppressive eosinophils form galectin-10-containing synapses with T cells, then release galectin-10 via plasma membrane disintegration and extracellular trap formation containing nuclear DNA and galectin-10. DNase I treatment to dissolve extracellular traps partially abrogates T cell suppression by eosinophils. Only CD16-expressing suppressive eosinophils (not conventional CD16neg eosinophils) form these synapses and EETs.","method":"Confocal microscopy, imaging flow cytometry, DNase I dissolution assay, [3H]-thymidine proliferation assay","journal":"Clinical and experimental immunology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — imaging plus functional DNase abrogation assay; single lab, two orthogonal methods establishing release mechanism","pmids":["33080067"],"is_preprint":false},{"year":2022,"finding":"Crystalline (insoluble) galectin-10 has significantly greater potency to induce inflammatory chemokine/cytokine release (IL-1β, IL-6, IL-8, TNF-α, GM-CSF) in primary human nasal epithelial cells and nasal polyp tissue compared to soluble galectin-10, demonstrating that the crystalline state determines the inflammatory potency of galectin-10.","method":"Recombinant protein engineering of crystalline vs. soluble Gal-10 states, cytokine/chemokine gene expression in primary cells and patient-derived nasal polyps","journal":"Nano letters","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct comparison of two states with gene expression readout in primary patient-derived cells; single lab, single method type","pmids":["35274950"],"is_preprint":false},{"year":2023,"finding":"Extracellular galectin-10 (released via EETosis) upregulates matrix metalloproteinase (MMP) production in normal human epidermal keratinocytes and dermal fibroblasts through activation of p38 MAPK, ERK, and JNK signaling pathways, potentially contributing to bullous pemphigoid blister formation.","method":"In vitro stimulation of NHEKs and NHDFs with recombinant galectin-10, real-time PCR, ELISA, Western blotting, signaling pathway inhibition assays","journal":"Journal of dermatological science","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — recombinant protein stimulation with pathway inhibition and multiple readouts; single lab","pmids":["37640566"],"is_preprint":false},{"year":2001,"finding":"Transcriptional regulation of galectin-10: a GC box (-44 to -50) is required for full promoter activity and for butyric acid-induced upregulation; Sp1 binds the GC box and Oct1 binds the Oct site (-255 to -261). AML3 binds the AML site and YY1 binds the Inr sequence, both functioning as silencers in the galectin-10 promoter.","method":"Promoter deletion analysis, supershift EMSA, transcription factor binding assays","journal":"Life sciences","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — EMSA supershift with functional promoter dissection; single lab, multiple orthogonal methods","pmids":["11441910"],"is_preprint":false},{"year":2025,"finding":"Galectin-10 silencing in human eosinophils reduces expression of IL-4, IL-5, IL-8, MBP, and TNF-α but not ECP, and decreases phosphorylation of p38 and p65 (NF-κB), indicating that galectin-10 promotes eosinophilic inflammation via the p38 MAPK/NF-κB pathway.","method":"siRNA knockdown in human eosinophils, Western blotting for p-p38 and p-p65, ELISA for cytokines and granule proteins","journal":"Critical reviews in immunology","confidence":"Low","confidence_rationale":"Tier 3 / Weak — siRNA knockdown with ELISA/WB readout; single lab, single method approach without rescue or structural validation","pmids":["40921147"],"is_preprint":false},{"year":2025,"finding":"Gal-10 crystal formation is driven by charge-charge attractions at protein-protein interaction interfaces and is sensitive to pH changes and charged residue substitutions at packing interfaces. Arginine-rich peptides (R9 and R12Y8) dissolve gal-10 crystals by disrupting inter-protein charge interactions; intratracheal administration of R12Y8 in a murine gal-10 crystal-induced lung inflammation model reduced proinflammatory cytokine release and inflammatory cell infiltration.","method":"In vitro crystal dissolution assays, in vivo murine intratracheal model, cytokine measurement","journal":"ACS applied materials & interfaces","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — in vitro dissolution mechanistic study plus in vivo murine model; single lab, two orthogonal approaches","pmids":["39894983"],"is_preprint":false},{"year":2021,"finding":"Recombinant galectin-10 stimulates prostaglandin E2 production in oral keratinocytes and gingival fibroblasts, and induces IL-8, MMP-9, and C-reactive protein secretion in gingival fibroblasts; conditioned media from rGal-10-treated fibroblasts induced osteoclast differentiation.","method":"Recombinant protein stimulation of primary cells, ELISA, osteoclast differentiation assay","journal":"Molecular medicine reports","confidence":"Low","confidence_rationale":"Tier 3 / Weak — recombinant protein stimulation assays without pathway validation or mechanistic follow-up; single lab, single method type","pmids":["33300083"],"is_preprint":false}],"current_model":"CLC/Galectin-10 is an eosinophil-predominant galectin-family protein that forms a novel homodimer in which Glu33 from one subunit occludes the carbohydrate-binding site of the other, switching ligand specificity from β-galactosides to mannose/small hydroxylated molecules; it is stored in the peripheral cytoplasm (not granules) of eosinophils, is released extracellularly via EETosis (a ROS- and PAD4-dependent cytolytic cell death) rather than classical degranulation, interacts intracellularly with granule cationic RNases (EDN/ECP) to facilitate their vesicular packaging and transport, is essential for the suppressive and anergic functions of regulatory T cells and suppressive eosinophils (acting through immune synapses and extracellular traps to inhibit T cell proliferation), and in its crystalline state potently activates IL-1β-dependent and MAPK/NF-κB-driven inflammatory pathways in epithelial and stromal cells."},"narrative":{"mechanistic_narrative":"CLC/galectin-10 is an eosinophil-predominant galectin-family protein whose structural features redirect it from canonical β-galactoside binding toward a distinct ligand chemistry and a set of immune effector functions [PMID:10529229, PMID:29293962]. Despite carrying a carbohydrate recognition domain, it is not a lysophospholipase; rather it binds lysophospholipase inhibitors and physically associates with eosinophil lysophospholipases [PMID:11834744]. Structurally it forms a novel homodimer in which Glu33 from one subunit occludes the carbohydrate-binding site of the partner subunit, switching specificity away from disaccharides toward mannose and small hydroxylated molecules, with His53 central to ligand contact and Trp72/Trp127 governing ligand binding, dimerization, and nuclear transport [PMID:10529229, PMID:29293962, PMID:30239701]. In eosinophils it is stored in the peripheral cytoplasm rather than secretory granules and is not released by classical degranulation; instead it is liberated through EETosis, a ROS- and PAD4-dependent cytolytic cell death that deposits galectin-10 in extracellular traps [PMID:32108369, PMID:33750029]. Intracellularly it interacts with granule cationic ribonucleases EDN/RNS2 and ECP/RNS3 in a glycosylation-independent, non-inhibitory manner and is required for eosinophil granulogenesis [PMID:31982451]. Functionally, galectin-10 is essential for the anergy and suppressive activity of regulatory T cells and suppressive eosinophils, acting through immune synapses and extracellular traps to inhibit T cell proliferation [PMID:17502455, PMID:28515279, PMID:33080067], and in its crystalline state it potently drives IL-1β-, MAPK-, and NF-κB-dependent inflammatory responses in epithelial and stromal cells [PMID:35274950, PMID:37640566].","teleology":[{"year":1999,"claim":"Established that despite being a galectin, CLC/Gal-10 does not bind β-galactosides but selectively binds mannose, redefining its ligand chemistry through an altered CRD topology.","evidence":"X-ray crystallography of the CLC–mannose complex at 1.8 Å","pmids":["10529229"],"confidence":"High","gaps":["Biological ligand in vivo not identified","Did not address dimerization-dependent occlusion of the site"]},{"year":2001,"claim":"Defined the transcriptional control of galectin-10, identifying activating and silencing promoter elements and their binding factors.","evidence":"Promoter deletion analysis and supershift EMSA","pmids":["11441910"],"confidence":"Medium","gaps":["Did not link transcriptional regulation to eosinophil lineage signals","No in vivo validation"]},{"year":2002,"claim":"Resolved the long-standing 'Charcot-Leyden crystal = lysophospholipase' assumption by showing CLC has no intrinsic lysophospholipase activity but instead binds inhibitors and the actual enzymes.","evidence":"Antibody depletion enzyme assays, crystallography of CLC–inhibitor complexes, ligand blotting","pmids":["11834744"],"confidence":"High","gaps":["Functional consequence of lysophospholipase binding unclear","True endogenous physiological activity not established"]},{"year":2007,"claim":"Identified galectin-10 as a Treg-restricted intracellular protein required for their anergy and suppressive function, extending its role beyond eosinophils.","evidence":"Differential proteomics, flow cytometry, specific inhibition in human Tregs","pmids":["17502455"],"confidence":"Medium","gaps":["Molecular mechanism of suppression not defined","Intracellular target/partner in Tregs unknown"]},{"year":2017,"claim":"Showed eosinophil galectin-10 is a T cell-suppressive effector that acts via immune synapses and is sufficient as a recombinant protein.","evidence":"Antibody neutralization in coculture, recombinant protein assays, immunofluorescence of synapses","pmids":["28515279"],"confidence":"Medium","gaps":["Receptor on T cells unidentified","Suppression only partially abrogated by neutralization"]},{"year":2018,"claim":"Determined that galectin-10 forms a distinct homodimer in which Glu33 occludes the partner's binding site, mechanistically explaining its switched ligand specificity.","evidence":"Crystallography of Gal-10 and eight variants, SEC, hemagglutination, alanine mutagenesis","pmids":["29293962"],"confidence":"High","gaps":["Physiological relevance of mannose/glycerol binding unresolved","Dynamics of monomer-dimer equilibrium in cells unknown"]},{"year":2019,"claim":"Confirmed dimerization-dependent suppression of carbohydrate binding and assigned Trp127 to dimer interface and Trp72 to nuclear transport.","evidence":"Crystallography of E33A, binding assays, EGFP-tagged localization in HeLa, mutagenesis","pmids":["30239701"],"confidence":"High","gaps":["Functional purpose of nuclear localization not established","Endogenous trigger for monomer/dimer switching unknown"]},{"year":2020,"claim":"Defined galectin-10's intracellular partnership with eosinophil cationic RNases and its requirement for granulogenesis, linking it to granule biogenesis.","evidence":"Reciprocal co-IP/affinity purification, shRNA knockdown in CD34+ progenitor model, confocal imaging","pmids":["31982451"],"confidence":"High","gaps":["Mechanism by which it aids RNase packaging unclear","Whether interaction is direct or bridged not resolved"]},{"year":2020,"claim":"Established that galectin-10 resides in the peripheral cytoplasm and is not exported by classical degranulation, reframing its secretion route.","evidence":"Immunonanogold transmission EM with quantitative analysis, immunofluorescence, CCL11/TNF-α stimulation","pmids":["32108369"],"confidence":"High","gaps":["Mechanism anchoring it to plasma membrane microdomains unknown","Did not directly observe the release event"]},{"year":2020,"claim":"Showed that CD16+ suppressive eosinophils release galectin-10 via extracellular trap formation to suppress T cells, identifying a DNA-dependent suppressive mechanism.","evidence":"Confocal microscopy, imaging flow cytometry, DNase I dissolution, thymidine proliferation assay","pmids":["33080067"],"confidence":"Medium","gaps":["Suppression only partially DNase-sensitive","Eosinophil subset markers driving this not fully defined"]},{"year":2021,"claim":"Identified EETosis as the ROS- and PAD4-dependent cytolytic mechanism for galectin-10 release and tied it to disease tissue.","evidence":"In vitro EETosis with ROS/PAD4 inhibitors, EM and immunostaining of EGPA tissue, ELISA","pmids":["33750029"],"confidence":"High","gaps":["Upstream triggers of EETosis in disease unclear","Fate of released galectin-10 in tissue not traced"]},{"year":2022,"claim":"Demonstrated that the crystalline state, not merely presence, of galectin-10 dictates its inflammatory potency in airway epithelium.","evidence":"Engineered crystalline vs soluble Gal-10, cytokine gene expression in primary cells and nasal polyps","pmids":["35274950"],"confidence":"Medium","gaps":["Receptor/sensor recognizing crystals unidentified","In vivo relevance beyond patient tissue not tested here"]},{"year":2023,"claim":"Showed extracellular galectin-10 activates p38/ERK/JNK signaling to upregulate MMPs in skin cells, extending its pro-inflammatory reach.","evidence":"Recombinant Gal-10 stimulation of keratinocytes/fibroblasts, qPCR, ELISA, WB, pathway inhibition","pmids":["37640566"],"confidence":"Medium","gaps":["Cell-surface receptor mediating MAPK activation unknown","Direct causal link to blister formation not demonstrated"]},{"year":2025,"claim":"Defined the physicochemical basis of galectin-10 crystallization and demonstrated therapeutic crystal dissolution in vivo.","evidence":"In vitro crystal dissolution assays, murine intratracheal crystal-induced inflammation model","pmids":["39894983"],"confidence":"Medium","gaps":["Specificity of arginine-rich peptides in vivo not fully characterized","Long-term efficacy/safety not assessed"]},{"year":2025,"claim":"Implicated galectin-10 in promoting eosinophilic inflammation through p38 MAPK/NF-κB signaling and cytokine expression.","evidence":"siRNA knockdown in human eosinophils, WB for p-p38/p-p65, ELISA","pmids":["40921147"],"confidence":"Low","gaps":["Single-lab siRNA approach without rescue or structural validation","Direct vs indirect effect on signaling not separated"]},{"year":null,"claim":"The cell-surface receptor(s) mediating galectin-10's extracellular T cell-suppressive and pro-inflammatory effects, and the trigger that governs its monomer/dimer and soluble/crystalline transitions, remain unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No receptor identified for extracellular galectin-10","Endogenous control of crystallization in vivo undefined","Connection between Treg-intrinsic and eosinophil-secreted functions unclear"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[2,8]}],"localization":[{"term_id":"GO:0005829","term_label":"cytosol","supporting_discovery_ids":[6]},{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[6]},{"term_id":"GO:0005634","term_label":"nucleus","supporting_discovery_ids":[4]},{"term_id":"GO:0005576","term_label":"extracellular region","supporting_discovery_ids":[7]}],"pathway":[{"term_id":"R-HSA-168256","term_label":"Immune System","supporting_discovery_ids":[2,8,9]},{"term_id":"R-HSA-5357801","term_label":"Programmed Cell Death","supporting_discovery_ids":[7]}],"complexes":[],"partners":["RNASE2","RNASE3"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q05315","full_name":"Galectin-10","aliases":["Charcot-Leyden crystal protein","CLC","Eosinophil lysophospholipase","Lysolecithin acylhydrolase"],"length_aa":142,"mass_kda":16.5,"function":"Regulates immune responses through the recognition of cell-surface glycans. Essential for the anergy and suppressive function of CD25-positive regulatory T-cells (Treg)","subcellular_location":"Cytoplasm, cytosol; Cytoplasmic granule","url":"https://www.uniprot.org/uniprotkb/Q05315/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/CLC","classification":"Not Classified","n_dependent_lines":3,"n_total_lines":1208,"dependency_fraction":0.0024834437086092716},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/CLC","total_profiled":1310},"omim":[{"mim_id":"619517","title":"NEURODEVELOPMENTAL DISORDER WITH SEIZURES AND BRAIN ABNORMALITIES; NEDSBA","url":"https://www.omim.org/entry/619517"},{"mim_id":"619173","title":"CEROID LIPOFUSCINOSIS, NEURONAL, 15; CLN15","url":"https://www.omim.org/entry/619173"},{"mim_id":"615651","title":"LEUKOENCEPHALOPATHY WITH ATAXIA; LKPAT","url":"https://www.omim.org/entry/615651"},{"mim_id":"613939","title":"SPERMATOGENESIS-ASSOCIATED PROTEIN 20; SPATA20","url":"https://www.omim.org/entry/613939"},{"mim_id":"611490","title":"OSTEOPETROSIS,  AUTOSOMAL RECESSIVE 4; OPTB4","url":"https://www.omim.org/entry/611490"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"","locations":[],"tissue_specificity":"Tissue enriched","tissue_distribution":"Detected in many","driving_tissues":[{"tissue":"bone marrow","ntpm":426.7}],"url":"https://www.proteinatlas.org/search/CLC"},"hgnc":{"alias_symbol":["LGALS10","MGC149659","Gal-10"],"prev_symbol":[]},"alphafold":{"accession":"Q05315","domains":[{"cath_id":"2.60.120.200","chopping":"9-140","consensus_level":"high","plddt":97.907,"start":9,"end":140}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q05315","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q05315-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q05315-F1-predicted_aligned_error_v6.png","plddt_mean":97.06},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=CLC","jax_strain_url":"https://www.jax.org/strain/search?query=CLC"},"sequence":{"accession":"Q05315","fasta_url":"https://rest.uniprot.org/uniprotkb/Q05315.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q05315/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q05315"}},"corpus_meta":[{"pmid":"17502455","id":"PMC_17502455","title":"Human CD4+CD25+ regulatory T cells: proteome analysis identifies galectin-10 as a novel marker essential for their anergy and suppressive function.","date":"2007","source":"Blood","url":"https://pubmed.ncbi.nlm.nih.gov/17502455","citation_count":162,"is_preprint":false},{"pmid":"11834744","id":"PMC_11834744","title":"Charcot-Leyden crystal protein (galectin-10) is not a dual function galectin with lysophospholipase activity but binds a lysophospholipase inhibitor in a novel structural fashion.","date":"2002","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/11834744","citation_count":94,"is_preprint":false},{"pmid":"10529229","id":"PMC_10529229","title":"Selective recognition of mannose by the human eosinophil Charcot-Leyden crystal protein (galectin-10): a crystallographic study at 1.8 A resolution.","date":"1999","source":"Biochemistry","url":"https://pubmed.ncbi.nlm.nih.gov/10529229","citation_count":74,"is_preprint":false},{"pmid":"33750029","id":"PMC_33750029","title":"Eosinophil ETosis-Mediated Release of Galectin-10 in Eosinophilic Granulomatosis With Polyangiitis.","date":"2021","source":"Arthritis & rheumatology (Hoboken, N.J.)","url":"https://pubmed.ncbi.nlm.nih.gov/33750029","citation_count":71,"is_preprint":false},{"pmid":"30424011","id":"PMC_30424011","title":"A Brief History of Charcot-Leyden Crystal Protein/Galectin-10 Research.","date":"2018","source":"Molecules (Basel, Switzerland)","url":"https://pubmed.ncbi.nlm.nih.gov/30424011","citation_count":57,"is_preprint":false},{"pmid":"28515279","id":"PMC_28515279","title":"Regulatory Eosinophils Suppress T Cells Partly through Galectin-10.","date":"2017","source":"Journal of immunology (Baltimore, Md. : 1950)","url":"https://pubmed.ncbi.nlm.nih.gov/28515279","citation_count":56,"is_preprint":false},{"pmid":"32108369","id":"PMC_32108369","title":"Galectin-10, the protein that forms Charcot-Leyden crystals, is not stored in granules but resides in the peripheral cytoplasm of human eosinophils.","date":"2020","source":"Journal of leukocyte biology","url":"https://pubmed.ncbi.nlm.nih.gov/32108369","citation_count":42,"is_preprint":false},{"pmid":"31982451","id":"PMC_31982451","title":"Charcot-Leyden crystal protein/galectin-10 interacts with cationic ribonucleases and is required for eosinophil granulogenesis.","date":"2020","source":"The Journal of allergy and clinical immunology","url":"https://pubmed.ncbi.nlm.nih.gov/31982451","citation_count":38,"is_preprint":false},{"pmid":"36291593","id":"PMC_36291593","title":"Galectin-10 as a Potential Biomarker for Eosinophilic 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allergy","url":"https://pubmed.ncbi.nlm.nih.gov/34647485","citation_count":33,"is_preprint":false},{"pmid":"29293962","id":"PMC_29293962","title":"Galectin-10: a new structural type of prototype galectin dimer and effects on saccharide ligand binding.","date":"2018","source":"Glycobiology","url":"https://pubmed.ncbi.nlm.nih.gov/29293962","citation_count":31,"is_preprint":false},{"pmid":"19758173","id":"PMC_19758173","title":"Galectin-10, eosinophils, and celiac disease.","date":"2009","source":"Annals of the New York Academy of Sciences","url":"https://pubmed.ncbi.nlm.nih.gov/19758173","citation_count":27,"is_preprint":false},{"pmid":"33080067","id":"PMC_33080067","title":"Kinetic studies of galectin-10 release from eosinophils exposed to proliferating T cells.","date":"2020","source":"Clinical and experimental immunology","url":"https://pubmed.ncbi.nlm.nih.gov/33080067","citation_count":21,"is_preprint":false},{"pmid":"34386407","id":"PMC_34386407","title":"Mepolizumab decreased the levels of serum galectin-10 and eosinophil cationic protein in asthma.","date":"2021","source":"Asia Pacific allergy","url":"https://pubmed.ncbi.nlm.nih.gov/34386407","citation_count":19,"is_preprint":false},{"pmid":"30239701","id":"PMC_30239701","title":"Identification of key amino acid residues determining ligand binding specificity, homodimerization and cellular distribution of human galectin-10.","date":"2019","source":"Glycobiology","url":"https://pubmed.ncbi.nlm.nih.gov/30239701","citation_count":17,"is_preprint":false},{"pmid":"11441910","id":"PMC_11441910","title":"Transcriptional regulation of galectin-10 (eosinophil Charcot-Leyden crystal protein): a GC box (-44 to -50) controls butyric acid induction of gene expression.","date":"2001","source":"Life sciences","url":"https://pubmed.ncbi.nlm.nih.gov/11441910","citation_count":17,"is_preprint":false},{"pmid":"22654612","id":"PMC_22654612","title":"Galectin-10 is released in the nasal lavage fluid of patients with aspirin-sensitive respiratory disease.","date":"2012","source":"TheScientificWorldJournal","url":"https://pubmed.ncbi.nlm.nih.gov/22654612","citation_count":16,"is_preprint":false},{"pmid":"38551536","id":"PMC_38551536","title":"Galectin-10 in serum extracellular vesicles reflects asthma pathophysiology.","date":"2024","source":"The Journal of allergy and clinical immunology","url":"https://pubmed.ncbi.nlm.nih.gov/38551536","citation_count":15,"is_preprint":false},{"pmid":"33300083","id":"PMC_33300083","title":"Identification of galectin‑10 as a biomarker for periodontitis based on proteomic analysis of gingival crevicular fluid.","date":"2020","source":"Molecular medicine reports","url":"https://pubmed.ncbi.nlm.nih.gov/33300083","citation_count":11,"is_preprint":false},{"pmid":"23224730","id":"PMC_23224730","title":"The mRNA level of the galectin-10 of Angiostrongylus cantonensis induced by reactive oxygen stress.","date":"2012","source":"Parasitology research","url":"https://pubmed.ncbi.nlm.nih.gov/23224730","citation_count":11,"is_preprint":false},{"pmid":"37640566","id":"PMC_37640566","title":"Eosinophil-derived galectin-10 upregulates matrix metalloproteinase expression in bullous pemphigoid blisters.","date":"2023","source":"Journal of dermatological science","url":"https://pubmed.ncbi.nlm.nih.gov/37640566","citation_count":9,"is_preprint":false},{"pmid":"35274950","id":"PMC_35274950","title":"Crystalline State Determines the Potency of Galectin-10 Protein Assembly to Induce Inflammation.","date":"2022","source":"Nano letters","url":"https://pubmed.ncbi.nlm.nih.gov/35274950","citation_count":9,"is_preprint":false},{"pmid":"20858065","id":"PMC_20858065","title":"The mRNA level of Charcot-Leyden crystal protein/galectin-10 is a marker for CRTH2 activation in human whole blood in vitro.","date":"2010","source":"Biomarkers : biochemical indicators of exposure, response, and susceptibility to chemicals","url":"https://pubmed.ncbi.nlm.nih.gov/20858065","citation_count":9,"is_preprint":false},{"pmid":"37998731","id":"PMC_37998731","title":"Galectin-10 Expression in Placentas of Women with Gestational Diabetes.","date":"2023","source":"Current issues in molecular biology","url":"https://pubmed.ncbi.nlm.nih.gov/37998731","citation_count":8,"is_preprint":false},{"pmid":"36808213","id":"PMC_36808213","title":"Extracellular distribution of galectin-10 in the esophageal mucosa of patients with eosinophilic esophagitis.","date":"2023","source":"Clinical and experimental immunology","url":"https://pubmed.ncbi.nlm.nih.gov/36808213","citation_count":5,"is_preprint":false},{"pmid":"33719171","id":"PMC_33719171","title":"A potential contribution of decreased serum galectin-10 levels to systemic inflammation and pulmonary vascular involvement in systemic sclerosis.","date":"2021","source":"Experimental dermatology","url":"https://pubmed.ncbi.nlm.nih.gov/33719171","citation_count":3,"is_preprint":false},{"pmid":"32911886","id":"PMC_32911886","title":"[Expression and pathological role of galectin-10 in different types of nasal polyps].","date":"2020","source":"Zhonghua er bi yan hou tou jing wai ke za zhi = Chinese journal of otorhinolaryngology head and neck surgery","url":"https://pubmed.ncbi.nlm.nih.gov/32911886","citation_count":2,"is_preprint":false},{"pmid":"40822235","id":"PMC_40822235","title":"C-BIOPRED severe asthma clinical phenotypes: link to complement and coagulation pathways and galectin 10.","date":"2025","source":"ERJ open research","url":"https://pubmed.ncbi.nlm.nih.gov/40822235","citation_count":1,"is_preprint":false},{"pmid":"40921147","id":"PMC_40921147","title":"Galectin-10 Silencing Reduces Eosinophilic Inflammation in Chronic Rhinosinusitis with Nasal Polyps by Inhibiting the p38/MAPK/NF-κB Pathway.","date":"2025","source":"Critical reviews in immunology","url":"https://pubmed.ncbi.nlm.nih.gov/40921147","citation_count":1,"is_preprint":false},{"pmid":"39252790","id":"PMC_39252790","title":"Serum Galectin-10: A biomarker for persistent airflow limitation in adult asthmatics.","date":"2024","source":"The World Allergy Organization journal","url":"https://pubmed.ncbi.nlm.nih.gov/39252790","citation_count":1,"is_preprint":false},{"pmid":"39894983","id":"PMC_39894983","title":"Arginine-Rich Peptides Regulate the Pathogenic Galectin-10 Crystallization and Mitigate Crystallopathy-Associated Inflammation.","date":"2025","source":"ACS applied materials & interfaces","url":"https://pubmed.ncbi.nlm.nih.gov/39894983","citation_count":1,"is_preprint":false},{"pmid":"41226378","id":"PMC_41226378","title":"Plasma Galectin-4 and Charcot-Leyden Crystal Protein/Galectin-10 as Emerging Biomarkers of Metabolically Induced Inflammation in Patients with Psoriasis.","date":"2025","source":"International journal of molecular sciences","url":"https://pubmed.ncbi.nlm.nih.gov/41226378","citation_count":1,"is_preprint":false},{"pmid":"41553873","id":"PMC_41553873","title":"Salivary galectin-7, galectin-10, and MMP-9 levels in periodontally healthy, gingivitis, and periodontitis patients.","date":"2026","source":"Journal of periodontology","url":"https://pubmed.ncbi.nlm.nih.gov/41553873","citation_count":0,"is_preprint":false},{"pmid":"41594617","id":"PMC_41594617","title":"Crystalline Insights into Nasal Mucosa Inflammation and Remodeling: Unveiling Role of Galectin-10.","date":"2026","source":"Biomolecules","url":"https://pubmed.ncbi.nlm.nih.gov/41594617","citation_count":0,"is_preprint":false},{"pmid":"40641544","id":"PMC_40641544","title":"Galectin-10 Characterization in Cleft Lip Palate - Affected Palatal Tissue.","date":"2025","source":"Acta medica Lituanica","url":"https://pubmed.ncbi.nlm.nih.gov/40641544","citation_count":0,"is_preprint":false},{"pmid":"41808620","id":"PMC_41808620","title":"Structurally Resilient Peptide Assembly Regulates Pathogenic Galectin-10 Crystallization To Mitigate Crystallopathy Inflammation.","date":"2026","source":"Journal of the American Chemical Society","url":"https://pubmed.ncbi.nlm.nih.gov/41808620","citation_count":0,"is_preprint":false}],"cost":{"stage1":{"sonnet_model":"claude-sonnet-4-6","sonnet_input_tokens":22760,"sonnet_output_tokens":4531,"sonnet_usd":0.1362,"sonnet_elapsed_seconds":83.5,"stage1_stop_reason":"end_turn"},"stage2":{"opus_model":"claude-opus-4-8","opus_input_tokens":12420,"opus_output_tokens":4125,"opus_usd":0.1652,"opus_elapsed_seconds":45.7,"stage2_stop_reason":"end_turn"},"total_usd":0.3014},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2002,\n      \"finding\": \"CLC protein (galectin-10) is NOT a lysophospholipase: antibody depletion of CLC from eosinophil lysates retained full lysophospholipase activity, and purified CLC protein lacked significant lysophospholipase activity. Instead, CLC protein binds lysophospholipase inhibitors (p-chloromercuribenzenesulfonate via disulfide bonds with Cys29 and Cys57, and N-ethylmaleimide via ring stacking with Trp72 in the carbohydrate recognition domain) and physically interacts with eosinophil lysophospholipases.\",\n      \"method\": \"Antibody affinity chromatography depletion, enzyme activity assays, X-ray crystallography of CLC–inhibitor complexes, ligand blotting\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — X-ray crystal structures plus enzymatic depletion assays plus ligand blotting; multiple orthogonal methods definitively establishing both a negative and a positive mechanistic finding in a single rigorous study\",\n      \"pmids\": [\"11834744\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1999,\n      \"finding\": \"CLC protein (galectin-10) selectively binds mannose (not β-galactosides) through its carbohydrate recognition domain (CRD); partial conservation of galectin CRD residues results in altered topology and chemistry of the binding site, explaining the switch in carbohydrate specificity.\",\n      \"method\": \"X-ray crystallography of CLC–mannose complex at 1.8 Å resolution\",\n      \"journal\": \"Biochemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — crystal structure with direct ligand complex at high resolution; single rigorous study with structural validation\",\n      \"pmids\": [\"10529229\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"Galectin-10 is constitutively and exclusively expressed intracellularly in human CD4+CD25+Foxp3+ regulatory T cells (Tregs) but not in resting or activated conventional CD4+ T cells; specific inhibition of galectin-10 restored proliferative capacity of Tregs and abrogated their suppressive function, establishing galectin-10 as essential for Treg anergy and suppression.\",\n      \"method\": \"Differential proteomics, single-cell staining, flow cytometry, specific inhibition of galectin-10\",\n      \"journal\": \"Blood\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — proteomics identification confirmed at mRNA and protein levels, functional inhibition with defined phenotypic readout; single lab with multiple orthogonal methods\",\n      \"pmids\": [\"17502455\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"Galectin-10 forms a novel homodimer with a global shape distinct from other prototype galectins (Gal-1, -2, -7); in the dimer, Glu33 from one subunit blocks the carbohydrate-binding site of the other subunit, inhibiting disaccharide binding. Glycerol and small hydroxylated molecules bind the ligand-binding site with His53 being most crucial. Trp72 negatively regulates ligand binding (W72A mutant shows enhanced erythrocyte agglutination).\",\n      \"method\": \"X-ray crystallography of Gal-10 and eight variants (1.55–2.00 Å resolution), size exclusion chromatography, hemagglutination assay, alanine mutagenesis\",\n      \"journal\": \"Glycobiology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — multiple crystal structures plus mutagenesis plus biochemical assays in one study; multiple orthogonal methods\",\n      \"pmids\": [\"29293962\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"Glu33 in the galectin-10 dimer interface blocks lactose binding; E33A mutant adopts a conformation allowing lactose binding as shown structurally and biochemically. Trp127 at the homodimer interface is essential for dimerization; the W127A monomer shows higher hemagglutination activity and can bind lactose-modified sepharose, confirming that dimerization suppresses carbohydrate binding. Trp72 is required for nuclear transport of Gal-10 (EGFP-tagged W72A cannot be transported into the nucleus in HeLa cells).\",\n      \"method\": \"X-ray crystallography of E33A variant, hemagglutination assay, solid-phase binding assay, EGFP-tagged subcellular localization in HeLa cells, site-directed mutagenesis\",\n      \"journal\": \"Glycobiology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — structural plus biochemical plus cell imaging with mutagenesis; multiple orthogonal methods in one study\",\n      \"pmids\": [\"30239701\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"CLC/Gal-10 interacts with eosinophil granule cationic ribonucleases (eosinophil-derived neurotoxin/RNS2 and eosinophil cationic protein/RNS3) and with murine eosinophil-associated RNases; this interaction is independent of glycosylation and is not inhibitory to endoRNase activity. Knockdown of CLC/Gal-10 in human CD34+ cord blood-derived progenitor eosinophils impairs eosinophil granulogenesis. IFN-γ activation induces rapid colocalization of CLC/Gal-10 with EDN/RNS2 and CD63, suggesting a role in vesicular transport of cationic RNases during degranulation.\",\n      \"method\": \"Ligand blotting, co-immunoprecipitation, co-affinity purification, enzyme activity assays, confocal microscopy, shRNA knockdown in CD34+ progenitor differentiation model\",\n      \"journal\": \"The Journal of allergy and clinical immunology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal co-IP/affinity purification plus shRNA KD with defined granulogenesis phenotype plus confocal imaging; multiple orthogonal methods\",\n      \"pmids\": [\"31982451\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"CLC-P/Gal-10 is stored predominantly in the peripheral cytoplasm of human eosinophils (within ~250 nm of the plasma membrane) and NOT within secretory (specific) granules; stimulation with CCL11 or TNF-α does not change this peripheral localization, indicating CLC-P/Gal-10 is not exported through classical degranulation (piecemeal degranulation or compound exocytosis). High-density microdomains of CLC-P/Gal-10 interact with the plasma membrane in ~60% of the membrane area.\",\n      \"method\": \"Pre-embedding immunonanogold transmission electron microscopy, quantitative imaging analysis, immunofluorescence, eosinophil activation with CCL11 and TNF-α\",\n      \"journal\": \"Journal of leukocyte biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — high-resolution immuno-EM with quantitative analysis plus immunofluorescence; orthogonal localization methods with functional context\",\n      \"pmids\": [\"32108369\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"EETosis (eosinophil extracellular trap cell death) is the mechanism for galectin-10 release from eosinophils; EETosis is dependent on reactive oxygen species and PAD4-dependent histone citrullination, resulting in cytolytic release of net-like extracellular traps containing galectin-10, free galectin-10, and membrane-bound intact granules. Loss of cytoplasmic galectin-10 and extracellular deposition are signatures of EETosis in eosinophilic granulomatosis with polyangiitis (EGPA) tissues.\",\n      \"method\": \"In vitro eosinophil EETosis characterization, ROS and PAD4 inhibition assays, immunostaining, electron microscopy of EGPA tissues, ELISA\",\n      \"journal\": \"Arthritis & rheumatology (Hoboken, N.J.)\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — in vitro mechanistic dissection with inhibitors plus EM and immunostaining in patient tissues; multiple orthogonal methods\",\n      \"pmids\": [\"33750029\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"Galectin-10 functions as a T cell-suppressive molecule in eosinophils: antibody-mediated neutralization of galectin-10 partially abrogated eosinophil-mediated suppression of T cell proliferation; recombinant galectin-10 alone suppressed T cell proliferation. Galectin-10-containing immune synapses form between eosinophils and lymphocytes.\",\n      \"method\": \"Antibody neutralization in coculture, recombinant protein functional assay, FACS sorting, immunofluorescence imaging of immune synapses\",\n      \"journal\": \"Journal of immunology (Baltimore, Md. : 1950)\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — neutralization plus recombinant protein functional assay plus imaging; single lab with multiple orthogonal methods\",\n      \"pmids\": [\"28515279\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"CD16+ suppressive eosinophils form galectin-10-containing synapses with T cells, then release galectin-10 via plasma membrane disintegration and extracellular trap formation containing nuclear DNA and galectin-10. DNase I treatment to dissolve extracellular traps partially abrogates T cell suppression by eosinophils. Only CD16-expressing suppressive eosinophils (not conventional CD16neg eosinophils) form these synapses and EETs.\",\n      \"method\": \"Confocal microscopy, imaging flow cytometry, DNase I dissolution assay, [3H]-thymidine proliferation assay\",\n      \"journal\": \"Clinical and experimental immunology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — imaging plus functional DNase abrogation assay; single lab, two orthogonal methods establishing release mechanism\",\n      \"pmids\": [\"33080067\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"Crystalline (insoluble) galectin-10 has significantly greater potency to induce inflammatory chemokine/cytokine release (IL-1β, IL-6, IL-8, TNF-α, GM-CSF) in primary human nasal epithelial cells and nasal polyp tissue compared to soluble galectin-10, demonstrating that the crystalline state determines the inflammatory potency of galectin-10.\",\n      \"method\": \"Recombinant protein engineering of crystalline vs. soluble Gal-10 states, cytokine/chemokine gene expression in primary cells and patient-derived nasal polyps\",\n      \"journal\": \"Nano letters\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct comparison of two states with gene expression readout in primary patient-derived cells; single lab, single method type\",\n      \"pmids\": [\"35274950\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"Extracellular galectin-10 (released via EETosis) upregulates matrix metalloproteinase (MMP) production in normal human epidermal keratinocytes and dermal fibroblasts through activation of p38 MAPK, ERK, and JNK signaling pathways, potentially contributing to bullous pemphigoid blister formation.\",\n      \"method\": \"In vitro stimulation of NHEKs and NHDFs with recombinant galectin-10, real-time PCR, ELISA, Western blotting, signaling pathway inhibition assays\",\n      \"journal\": \"Journal of dermatological science\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — recombinant protein stimulation with pathway inhibition and multiple readouts; single lab\",\n      \"pmids\": [\"37640566\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2001,\n      \"finding\": \"Transcriptional regulation of galectin-10: a GC box (-44 to -50) is required for full promoter activity and for butyric acid-induced upregulation; Sp1 binds the GC box and Oct1 binds the Oct site (-255 to -261). AML3 binds the AML site and YY1 binds the Inr sequence, both functioning as silencers in the galectin-10 promoter.\",\n      \"method\": \"Promoter deletion analysis, supershift EMSA, transcription factor binding assays\",\n      \"journal\": \"Life sciences\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — EMSA supershift with functional promoter dissection; single lab, multiple orthogonal methods\",\n      \"pmids\": [\"11441910\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"Galectin-10 silencing in human eosinophils reduces expression of IL-4, IL-5, IL-8, MBP, and TNF-α but not ECP, and decreases phosphorylation of p38 and p65 (NF-κB), indicating that galectin-10 promotes eosinophilic inflammation via the p38 MAPK/NF-κB pathway.\",\n      \"method\": \"siRNA knockdown in human eosinophils, Western blotting for p-p38 and p-p65, ELISA for cytokines and granule proteins\",\n      \"journal\": \"Critical reviews in immunology\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — siRNA knockdown with ELISA/WB readout; single lab, single method approach without rescue or structural validation\",\n      \"pmids\": [\"40921147\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"Gal-10 crystal formation is driven by charge-charge attractions at protein-protein interaction interfaces and is sensitive to pH changes and charged residue substitutions at packing interfaces. Arginine-rich peptides (R9 and R12Y8) dissolve gal-10 crystals by disrupting inter-protein charge interactions; intratracheal administration of R12Y8 in a murine gal-10 crystal-induced lung inflammation model reduced proinflammatory cytokine release and inflammatory cell infiltration.\",\n      \"method\": \"In vitro crystal dissolution assays, in vivo murine intratracheal model, cytokine measurement\",\n      \"journal\": \"ACS applied materials & interfaces\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vitro dissolution mechanistic study plus in vivo murine model; single lab, two orthogonal approaches\",\n      \"pmids\": [\"39894983\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"Recombinant galectin-10 stimulates prostaglandin E2 production in oral keratinocytes and gingival fibroblasts, and induces IL-8, MMP-9, and C-reactive protein secretion in gingival fibroblasts; conditioned media from rGal-10-treated fibroblasts induced osteoclast differentiation.\",\n      \"method\": \"Recombinant protein stimulation of primary cells, ELISA, osteoclast differentiation assay\",\n      \"journal\": \"Molecular medicine reports\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — recombinant protein stimulation assays without pathway validation or mechanistic follow-up; single lab, single method type\",\n      \"pmids\": [\"33300083\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"CLC/Galectin-10 is an eosinophil-predominant galectin-family protein that forms a novel homodimer in which Glu33 from one subunit occludes the carbohydrate-binding site of the other, switching ligand specificity from β-galactosides to mannose/small hydroxylated molecules; it is stored in the peripheral cytoplasm (not granules) of eosinophils, is released extracellularly via EETosis (a ROS- and PAD4-dependent cytolytic cell death) rather than classical degranulation, interacts intracellularly with granule cationic RNases (EDN/ECP) to facilitate their vesicular packaging and transport, is essential for the suppressive and anergic functions of regulatory T cells and suppressive eosinophils (acting through immune synapses and extracellular traps to inhibit T cell proliferation), and in its crystalline state potently activates IL-1β-dependent and MAPK/NF-κB-driven inflammatory pathways in epithelial and stromal cells.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"CLC/galectin-10 is an eosinophil-predominant galectin-family protein whose structural features redirect it from canonical β-galactoside binding toward a distinct ligand chemistry and a set of immune effector functions [#1, #3]. Despite carrying a carbohydrate recognition domain, it is not a lysophospholipase; rather it binds lysophospholipase inhibitors and physically associates with eosinophil lysophospholipases [#0]. Structurally it forms a novel homodimer in which Glu33 from one subunit occludes the carbohydrate-binding site of the partner subunit, switching specificity away from disaccharides toward mannose and small hydroxylated molecules, with His53 central to ligand contact and Trp72/Trp127 governing ligand binding, dimerization, and nuclear transport [#1, #3, #4]. In eosinophils it is stored in the peripheral cytoplasm rather than secretory granules and is not released by classical degranulation; instead it is liberated through EETosis, a ROS- and PAD4-dependent cytolytic cell death that deposits galectin-10 in extracellular traps [#6, #7]. Intracellularly it interacts with granule cationic ribonucleases EDN/RNS2 and ECP/RNS3 in a glycosylation-independent, non-inhibitory manner and is required for eosinophil granulogenesis [#5]. Functionally, galectin-10 is essential for the anergy and suppressive activity of regulatory T cells and suppressive eosinophils, acting through immune synapses and extracellular traps to inhibit T cell proliferation [#2, #8, #9], and in its crystalline state it potently drives IL-1β-, MAPK-, and NF-κB-dependent inflammatory responses in epithelial and stromal cells [#10, #11].\",\n  \"teleology\": [\n    {\n      \"year\": 1999,\n      \"claim\": \"Established that despite being a galectin, CLC/Gal-10 does not bind β-galactosides but selectively binds mannose, redefining its ligand chemistry through an altered CRD topology.\",\n      \"evidence\": \"X-ray crystallography of the CLC–mannose complex at 1.8 Å\",\n      \"pmids\": [\"10529229\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Biological ligand in vivo not identified\", \"Did not address dimerization-dependent occlusion of the site\"]\n    },\n    {\n      \"year\": 2001,\n      \"claim\": \"Defined the transcriptional control of galectin-10, identifying activating and silencing promoter elements and their binding factors.\",\n      \"evidence\": \"Promoter deletion analysis and supershift EMSA\",\n      \"pmids\": [\"11441910\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Did not link transcriptional regulation to eosinophil lineage signals\", \"No in vivo validation\"]\n    },\n    {\n      \"year\": 2002,\n      \"claim\": \"Resolved the long-standing 'Charcot-Leyden crystal = lysophospholipase' assumption by showing CLC has no intrinsic lysophospholipase activity but instead binds inhibitors and the actual enzymes.\",\n      \"evidence\": \"Antibody depletion enzyme assays, crystallography of CLC–inhibitor complexes, ligand blotting\",\n      \"pmids\": [\"11834744\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Functional consequence of lysophospholipase binding unclear\", \"True endogenous physiological activity not established\"]\n    },\n    {\n      \"year\": 2007,\n      \"claim\": \"Identified galectin-10 as a Treg-restricted intracellular protein required for their anergy and suppressive function, extending its role beyond eosinophils.\",\n      \"evidence\": \"Differential proteomics, flow cytometry, specific inhibition in human Tregs\",\n      \"pmids\": [\"17502455\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Molecular mechanism of suppression not defined\", \"Intracellular target/partner in Tregs unknown\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Showed eosinophil galectin-10 is a T cell-suppressive effector that acts via immune synapses and is sufficient as a recombinant protein.\",\n      \"evidence\": \"Antibody neutralization in coculture, recombinant protein assays, immunofluorescence of synapses\",\n      \"pmids\": [\"28515279\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Receptor on T cells unidentified\", \"Suppression only partially abrogated by neutralization\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Determined that galectin-10 forms a distinct homodimer in which Glu33 occludes the partner's binding site, mechanistically explaining its switched ligand specificity.\",\n      \"evidence\": \"Crystallography of Gal-10 and eight variants, SEC, hemagglutination, alanine mutagenesis\",\n      \"pmids\": [\"29293962\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Physiological relevance of mannose/glycerol binding unresolved\", \"Dynamics of monomer-dimer equilibrium in cells unknown\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Confirmed dimerization-dependent suppression of carbohydrate binding and assigned Trp127 to dimer interface and Trp72 to nuclear transport.\",\n      \"evidence\": \"Crystallography of E33A, binding assays, EGFP-tagged localization in HeLa, mutagenesis\",\n      \"pmids\": [\"30239701\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Functional purpose of nuclear localization not established\", \"Endogenous trigger for monomer/dimer switching unknown\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Defined galectin-10's intracellular partnership with eosinophil cationic RNases and its requirement for granulogenesis, linking it to granule biogenesis.\",\n      \"evidence\": \"Reciprocal co-IP/affinity purification, shRNA knockdown in CD34+ progenitor model, confocal imaging\",\n      \"pmids\": [\"31982451\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Mechanism by which it aids RNase packaging unclear\", \"Whether interaction is direct or bridged not resolved\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Established that galectin-10 resides in the peripheral cytoplasm and is not exported by classical degranulation, reframing its secretion route.\",\n      \"evidence\": \"Immunonanogold transmission EM with quantitative analysis, immunofluorescence, CCL11/TNF-α stimulation\",\n      \"pmids\": [\"32108369\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Mechanism anchoring it to plasma membrane microdomains unknown\", \"Did not directly observe the release event\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Showed that CD16+ suppressive eosinophils release galectin-10 via extracellular trap formation to suppress T cells, identifying a DNA-dependent suppressive mechanism.\",\n      \"evidence\": \"Confocal microscopy, imaging flow cytometry, DNase I dissolution, thymidine proliferation assay\",\n      \"pmids\": [\"33080067\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Suppression only partially DNase-sensitive\", \"Eosinophil subset markers driving this not fully defined\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Identified EETosis as the ROS- and PAD4-dependent cytolytic mechanism for galectin-10 release and tied it to disease tissue.\",\n      \"evidence\": \"In vitro EETosis with ROS/PAD4 inhibitors, EM and immunostaining of EGPA tissue, ELISA\",\n      \"pmids\": [\"33750029\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Upstream triggers of EETosis in disease unclear\", \"Fate of released galectin-10 in tissue not traced\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Demonstrated that the crystalline state, not merely presence, of galectin-10 dictates its inflammatory potency in airway epithelium.\",\n      \"evidence\": \"Engineered crystalline vs soluble Gal-10, cytokine gene expression in primary cells and nasal polyps\",\n      \"pmids\": [\"35274950\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Receptor/sensor recognizing crystals unidentified\", \"In vivo relevance beyond patient tissue not tested here\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Showed extracellular galectin-10 activates p38/ERK/JNK signaling to upregulate MMPs in skin cells, extending its pro-inflammatory reach.\",\n      \"evidence\": \"Recombinant Gal-10 stimulation of keratinocytes/fibroblasts, qPCR, ELISA, WB, pathway inhibition\",\n      \"pmids\": [\"37640566\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Cell-surface receptor mediating MAPK activation unknown\", \"Direct causal link to blister formation not demonstrated\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Defined the physicochemical basis of galectin-10 crystallization and demonstrated therapeutic crystal dissolution in vivo.\",\n      \"evidence\": \"In vitro crystal dissolution assays, murine intratracheal crystal-induced inflammation model\",\n      \"pmids\": [\"39894983\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Specificity of arginine-rich peptides in vivo not fully characterized\", \"Long-term efficacy/safety not assessed\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Implicated galectin-10 in promoting eosinophilic inflammation through p38 MAPK/NF-κB signaling and cytokine expression.\",\n      \"evidence\": \"siRNA knockdown in human eosinophils, WB for p-p38/p-p65, ELISA\",\n      \"pmids\": [\"40921147\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"Single-lab siRNA approach without rescue or structural validation\", \"Direct vs indirect effect on signaling not separated\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"The cell-surface receptor(s) mediating galectin-10's extracellular T cell-suppressive and pro-inflammatory effects, and the trigger that governs its monomer/dimer and soluble/crystalline transitions, remain unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No receptor identified for extracellular galectin-10\", \"Endogenous control of crystallization in vivo undefined\", \"Connection between Treg-intrinsic and eosinophil-secreted functions unclear\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [2, 8]},\n      {\"term_id\": \"GO:0030246\", \"supporting_discovery_ids\": [1, 3]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [6]},\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [6]},\n      {\"term_id\": \"GO:0005634\", \"supporting_discovery_ids\": [4]},\n      {\"term_id\": \"GO:0005576\", \"supporting_discovery_ids\": [7]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [2, 8, 9]},\n      {\"term_id\": \"R-HSA-5357801\", \"supporting_discovery_ids\": [7]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"RNASE2\", \"RNASE3\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"tie"}}