{"gene":"GNLY","run_date":"2026-04-28T18:06:53","timeline":{"discoveries":[{"year":1993,"finding":"The GNLY gene (NKG5) encodes a secreted protein expressed specifically in natural killer and T cells, strongly upregulated upon cell activation; the gene consists of five exons and four introns, and NKG5 and the related clone 519 are alternative splicing products of a single gene.","method":"Genomic library screening, sequencing, comparison with cDNA, and 5'-flanking region analysis","journal":"Immunogenetics","confidence":"Medium","confidence_rationale":"Tier 2 — direct genomic characterization with structural validation, single lab","pmids":["8423048"],"is_preprint":false},{"year":1990,"finding":"The GNLY gene (519/NKG5) was localized to human chromosome 2p12–q11 by in situ hybridization.","method":"In situ hybridization on human metaphase chromosomes","journal":"Cytogenetics and cell genetics","confidence":"Medium","confidence_rationale":"Tier 2 — direct localization experiment, single lab","pmids":["2209093"],"is_preprint":false},{"year":1999,"finding":"NKG5 (GNLY) protein stimulates mitogenicity of endothelial cells in vitro, suggesting a role in angiogenesis; it was identified in decidual extract fractions with hyperactive mitogenic activity at concentrations 1000-fold lower than bFGF.","method":"Heparin-sepharose column fractionation, endothelial cell proliferation assay (methylene blue stain), protein sequencing","journal":"American journal of reproductive immunology","confidence":"Low","confidence_rationale":"Tier 3 — single lab, single functional assay with protein identification but limited mechanistic follow-up","pmids":["10584980"],"is_preprint":false},{"year":2019,"finding":"A GNLY c.11G>A mutation creates a premature termination codon (p.Trp4Ter) associated with TEN; a mutant protein is synthesized (likely via PTC-readthrough) and is abnormally localized to the nuclear compartment, potentially leading to a toxic effect rather than the normal secretory pathway.","method":"Direct sequencing of GNLY coding region, functional localization assay","journal":"Human genetics","confidence":"Medium","confidence_rationale":"Tier 2 — direct mutagenesis with subcellular localization and functional consequence, single lab","pmids":["31642954"],"is_preprint":false},{"year":2025,"finding":"GNLY+CD8+ T cells predominantly express the pro-inflammatory 15 kDa form of granulysin and are activated by IL-15 in vitro; supernatant from IL-15-stimulated CD8+ T cells induces monocytes to secrete inflammatory factors and disrupts intestinal epithelial cell integrity, effects that can be partially restored by anti-GNLY antibodies.","method":"Single-cell RNA sequencing, in vitro IL-15 stimulation, anti-GNLY antibody neutralization assay, monocyte co-culture","journal":"Emerging microbes & infections","confidence":"Medium","confidence_rationale":"Tier 2 — multiple orthogonal methods (scRNA-seq, in vitro stimulation, antibody neutralization) in single study","pmids":["40135938"],"is_preprint":false}],"current_model":"GNLY (granulysin/NKG5) encodes a secreted antimicrobial and cytolytic peptide expressed in NK cells and cytotoxic T lymphocytes upon activation, encoded at chromosome 2p12-q11, produced as alternative splice forms including a pro-inflammatory 15 kDa form; granulysin is released from cytotoxic granules along with granzyme and perforin, can induce inflammatory signaling in monocytes and disrupt epithelial barrier integrity via a paracrine mechanism blocked by anti-GNLY antibodies, and proper N-terminal sequence is required for normal secretory localization since truncation mutants mislocalize to the nucleus."},"narrative":{"teleology":[{"year":1990,"claim":"Chromosomal mapping established that GNLY resides at 2p12–q11, placing it in a genomic context distinct from other cytolytic effector gene clusters and enabling subsequent cloning efforts.","evidence":"In situ hybridization on human metaphase chromosomes","pmids":["2209093"],"confidence":"Medium","gaps":["Gene structure and transcript diversity were not yet characterized","No functional data for the encoded protein"]},{"year":1993,"claim":"Cloning and structural analysis revealed that GNLY is a five-exon gene producing alternatively spliced transcripts (NKG5 and clone 519) encoding a secreted protein specifically upregulated in activated NK and T cells, establishing it as an activation-dependent immune effector gene.","evidence":"Genomic library screening, sequencing, 5′-flanking region analysis, and cDNA comparison","pmids":["8423048"],"confidence":"Medium","gaps":["Functional roles of the encoded protein (cytolysis, antimicrobial activity) were not tested","Regulation by specific cytokines was not defined"]},{"year":1999,"claim":"Identification of granulysin as a mitogenic factor for endothelial cells at sub-nanomolar concentrations raised the possibility that granulysin has paracrine signaling roles beyond direct cytolysis.","evidence":"Heparin-sepharose fractionation of decidual extract, endothelial proliferation assay, protein sequencing","pmids":["10584980"],"confidence":"Low","gaps":["Single functional assay without receptor identification or signaling pathway dissection","No in vivo validation of angiogenic activity","Not independently replicated"]},{"year":2019,"claim":"A premature termination mutation (p.Trp4Ter) demonstrated that the N-terminal signal peptide is essential for secretory pathway targeting; the truncated protein mislocalizes to the nucleus and is associated with toxic epidermal necrolysis, linking GNLY loss-of-function to severe adverse drug reactions.","evidence":"Direct sequencing of GNLY coding region and subcellular localization assay of mutant protein","pmids":["31642954"],"confidence":"Medium","gaps":["Mechanism of nuclear toxicity not elucidated","Single patient/family study — broader population confirmation needed","Whether PTC-readthrough product retains partial cytolytic activity is unknown"]},{"year":2025,"claim":"Mechanistic dissection showed that IL-15-activated CD8+ T cells produce the 15 kDa pro-inflammatory granulysin isoform, which acts in a paracrine manner to activate monocyte inflammatory responses and compromise intestinal epithelial barrier integrity — effects reversible by anti-GNLY antibodies — establishing granulysin as a mediator of tissue-damaging inflammation.","evidence":"Single-cell RNA-seq, IL-15 in vitro stimulation, monocyte co-culture, anti-GNLY antibody neutralization, epithelial barrier assays","pmids":["40135938"],"confidence":"Medium","gaps":["Receptor or signaling pathway on monocytes and epithelial cells not identified","In vivo relevance in gut inflammation not demonstrated","Relative contributions of 9 kDa vs 15 kDa isoforms to barrier disruption not resolved"]},{"year":null,"claim":"The receptor(s) through which granulysin signals on monocytes and epithelial cells, the structural basis of its membrane-disrupting activity against host versus microbial membranes, and the in vivo contribution of each splice isoform to immune defense versus tissue damage remain undefined.","evidence":"","pmids":[],"confidence":"Low","gaps":["No receptor identified for granulysin on target host cells","No structural model explaining selectivity for microbial vs host membranes","In vivo isoform-specific knockout studies lacking"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0048018","term_label":"receptor ligand activity","supporting_discovery_ids":[4]},{"term_id":"GO:0090729","term_label":"toxin activity","supporting_discovery_ids":[4]}],"localization":[{"term_id":"GO:0031410","term_label":"cytoplasmic vesicle","supporting_discovery_ids":[0,4]},{"term_id":"GO:0005576","term_label":"extracellular region","supporting_discovery_ids":[0,4]}],"pathway":[{"term_id":"R-HSA-168256","term_label":"Immune System","supporting_discovery_ids":[0,4]}],"complexes":[],"partners":[],"other_free_text":[]},"mechanistic_narrative":"GNLY encodes granulysin, a secreted cytolytic and pro-inflammatory peptide expressed in natural killer cells and cytotoxic T lymphocytes upon activation, produced as alternative splice forms from a five-exon gene on chromosome 2p12–q11 [PMID:8423048, PMID:2209093]. The 15 kDa pro-inflammatory form of granulysin, released from activated CD8+ T cells stimulated by IL-15, induces monocyte inflammatory cytokine secretion and disrupts intestinal epithelial barrier integrity via a paracellular mechanism that is blocked by anti-GNLY antibodies [PMID:40135938]. The N-terminal signal sequence is required for correct secretory pathway localization, as a premature termination mutation (p.Trp4Ter) yields a protein that mislocalizes to the nucleus with potential toxic consequences, linking GNLY to toxic epidermal necrolysis (TEN) [PMID:31642954]."},"prefetch_data":{"uniprot":{"accession":"P22749","full_name":"Granulysin","aliases":["Lymphokine LAG-2","Protein NKG5","T-cell activation protein 519"],"length_aa":145,"mass_kda":16.4,"function":"Antimicrobial protein that kills intracellular pathogens. Active against a broad range of microbes, including Gram-positive and Gram-negative bacteria, fungi, and parasites. Kills Mycobacterium tuberculosis","subcellular_location":"Secreted","url":"https://www.uniprot.org/uniprotkb/P22749/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/GNLY","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/GNLY","total_profiled":1310},"omim":[{"mim_id":"608579","title":"SEVERE CUTANEOUS ADVERSE REACTION, SUSCEPTIBILITY TO","url":"https://www.omim.org/entry/608579"},{"mim_id":"188855","title":"GRANULYSIN; GNLY","url":"https://www.omim.org/entry/188855"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"","locations":[],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in many","driving_tissues":[{"tissue":"bone marrow","ntpm":271.5},{"tissue":"endometrium 1","ntpm":67.6},{"tissue":"lymphoid tissue","ntpm":81.2}],"url":"https://www.proteinatlas.org/search/GNLY"},"hgnc":{"alias_symbol":["NKG5","LAG-2","D2S69E","TLA519"],"prev_symbol":["LAG2"]},"alphafold":{"accession":"P22749","domains":[{"cath_id":"1.10.225.10","chopping":"58-136","consensus_level":"high","plddt":94.8053,"start":58,"end":136}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/P22749","model_url":"https://alphafold.ebi.ac.uk/files/AF-P22749-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-P22749-F1-predicted_aligned_error_v6.png","plddt_mean":78.06},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=GNLY","jax_strain_url":"https://www.jax.org/strain/search?query=GNLY"},"sequence":{"accession":"P22749","fasta_url":"https://rest.uniprot.org/uniprotkb/P22749.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/P22749/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/P22749"}},"corpus_meta":[{"pmid":"7607081","id":"PMC_7607081","title":"lag-2 may 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gene.","date":"1993","source":"Immunogenetics","url":"https://pubmed.ncbi.nlm.nih.gov/8423048","citation_count":24,"is_preprint":false},{"pmid":"31090825","id":"PMC_31090825","title":"The role of GNLY gene polymorphisms in psoriasis pathogenesis.","date":"2019","source":"Anais brasileiros de dermatologia","url":"https://pubmed.ncbi.nlm.nih.gov/31090825","citation_count":16,"is_preprint":false},{"pmid":"19763088","id":"PMC_19763088","title":"A longevity protein, Lag2, interacts with SCF complex and regulates SCF function.","date":"2009","source":"The EMBO journal","url":"https://pubmed.ncbi.nlm.nih.gov/19763088","citation_count":15,"is_preprint":false},{"pmid":"35966395","id":"PMC_35966395","title":"An in vivo toolkit to visualize endogenous LAG-2/Delta and LIN-12/Notch signaling in C. elegans.","date":"2022","source":"microPublication biology","url":"https://pubmed.ncbi.nlm.nih.gov/35966395","citation_count":14,"is_preprint":false},{"pmid":"10584980","id":"PMC_10584980","title":"Novel hyperactive mitogen to endothelial cells: human decidual NKG5.","date":"1999","source":"American journal of reproductive immunology (New York, N.Y. : 1989)","url":"https://pubmed.ncbi.nlm.nih.gov/10584980","citation_count":14,"is_preprint":false},{"pmid":"22788687","id":"PMC_22788687","title":"Association of GNLY genetic polymorphisms with chronic liver disease in a Korean population.","date":"2012","source":"DNA and cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/22788687","citation_count":14,"is_preprint":false},{"pmid":"8760941","id":"PMC_8760941","title":"LAG2, a gene that determines yeast longevity.","date":"1996","source":"Microbiology (Reading, England)","url":"https://pubmed.ncbi.nlm.nih.gov/8760941","citation_count":12,"is_preprint":false},{"pmid":"35066979","id":"PMC_35066979","title":"GNLY gene polymorphism: A potential role in understanding psoriasis pathogenesis.","date":"2022","source":"Journal of cosmetic 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research","url":"https://pubmed.ncbi.nlm.nih.gov/39802509","citation_count":3,"is_preprint":false},{"pmid":"39437618","id":"PMC_39437618","title":"GNLY as A Novel Cis-eQTL and Cis-pQTL Mediated Susceptibility Gene in Suppressing Prostatitis. Mendelian Randomization Study.","date":"2024","source":"Archives of medical research","url":"https://pubmed.ncbi.nlm.nih.gov/39437618","citation_count":1,"is_preprint":false},{"pmid":"39675427","id":"PMC_39675427","title":"Spatially Resolved Single-Cell Transcriptome Analysis of Mycosis Fungoides Reveals Distinct Biomarkers GNLY and FYB1 Compared With Psoriasis and Chronic Spongiotic Dermatitis.","date":"2024","source":"Modern pathology : an official journal of the United States and Canadian Academy of Pathology, Inc","url":"https://pubmed.ncbi.nlm.nih.gov/39675427","citation_count":1,"is_preprint":false},{"pmid":null,"id":"bio_10.1101_2024.11.18.624213","title":"Dysregulated Proteins in Plasma Distinguishing Syndromic from Non-syndromic Heritable Thoracic Aortic Disease","date":"2024-11-22","source":"bioRxiv","url":"https://doi.org/10.1101/2024.11.18.624213","citation_count":0,"is_preprint":true},{"pmid":null,"id":"bio_10.1101_2025.05.23.655783","title":"Pre-dauer starvation rapidly and reversibly reduces niche proliferative signaling to the  <i>C. elegans</i>  germ line","date":"2025-05-28","source":"bioRxiv","url":"https://doi.org/10.1101/2025.05.23.655783","citation_count":0,"is_preprint":true},{"pmid":null,"id":"bio_10.1101_2025.03.25.644515","title":"Comprehensive longitudinal profiling of SARS-CoV-2-specific CD8<sup>+</sup>T-cells reveal strong functional impairment and recognition bias as markers for disease severity","date":"2025-03-26","source":"bioRxiv","url":"https://doi.org/10.1101/2025.03.25.644515","citation_count":0,"is_preprint":true},{"pmid":null,"id":"bio_10.1101_2025.02.03.636324","title":"Single-cell RNA-seq reveals altered plasma cell subsets and decreased cytotoxicity of NK cells in patients with Kawasaki disease","date":"2025-02-08","source":"bioRxiv","url":"https://doi.org/10.1101/2025.02.03.636324","citation_count":0,"is_preprint":true},{"pmid":null,"id":"bio_10.1101_2024.10.08.617199","title":"Proteomic and transcriptomic host biomarkers for detection of pleural tuberculosis","date":"2024-10-09","source":"bioRxiv","url":"https://doi.org/10.1101/2024.10.08.617199","citation_count":0,"is_preprint":true},{"pmid":null,"id":"bio_10.1101_2024.06.04.597327","title":"An organoid co-culture model for probing systemic anti-tumor immunity in lung cancer","date":"2024-06-06","source":"bioRxiv","url":"https://doi.org/10.1101/2024.06.04.597327","citation_count":0,"is_preprint":true},{"pmid":null,"id":"bio_10.1101_2024.05.08.593208","title":"The roles of TGFβ and serotonin signaling in regulating proliferation of oocyte precursors and germline aging","date":"2024-05-09","source":"bioRxiv","url":"https://doi.org/10.1101/2024.05.08.593208","citation_count":0,"is_preprint":true},{"pmid":null,"id":"bio_10.1101_2024.07.10.602225","title":"An in vivo screen for proteolytic switch domains that can mediate Notch activation by force","date":"2024-07-12","source":"bioRxiv","url":"https://doi.org/10.1101/2024.07.10.602225","citation_count":0,"is_preprint":true},{"pmid":null,"id":"bio_10.1101_2024.07.03.24309252","title":"Machine learning-based forecasting of daily acute ischemic stroke admissions using weather data","date":"2024-07-04","source":"bioRxiv","url":"https://doi.org/10.1101/2024.07.03.24309252","citation_count":0,"is_preprint":true}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":17119,"output_tokens":1135,"usd":0.034191},"stage2":{"model":"claude-opus-4-6","input_tokens":4354,"output_tokens":1556,"usd":0.091005},"total_usd":0.125196,"stage1_batch_id":"msgbatch_01DzrsgVgYngY1hvM3DU1yF2","stage2_batch_id":"msgbatch_01KA9b7iRzKFdhoBUT7vCvSe","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 1993,\n      \"finding\": \"The GNLY gene (NKG5) encodes a secreted protein expressed specifically in natural killer and T cells, strongly upregulated upon cell activation; the gene consists of five exons and four introns, and NKG5 and the related clone 519 are alternative splicing products of a single gene.\",\n      \"method\": \"Genomic library screening, sequencing, comparison with cDNA, and 5'-flanking region analysis\",\n      \"journal\": \"Immunogenetics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — direct genomic characterization with structural validation, single lab\",\n      \"pmids\": [\"8423048\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1990,\n      \"finding\": \"The GNLY gene (519/NKG5) was localized to human chromosome 2p12–q11 by in situ hybridization.\",\n      \"method\": \"In situ hybridization on human metaphase chromosomes\",\n      \"journal\": \"Cytogenetics and cell genetics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — direct localization experiment, single lab\",\n      \"pmids\": [\"2209093\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1999,\n      \"finding\": \"NKG5 (GNLY) protein stimulates mitogenicity of endothelial cells in vitro, suggesting a role in angiogenesis; it was identified in decidual extract fractions with hyperactive mitogenic activity at concentrations 1000-fold lower than bFGF.\",\n      \"method\": \"Heparin-sepharose column fractionation, endothelial cell proliferation assay (methylene blue stain), protein sequencing\",\n      \"journal\": \"American journal of reproductive immunology\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 — single lab, single functional assay with protein identification but limited mechanistic follow-up\",\n      \"pmids\": [\"10584980\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"A GNLY c.11G>A mutation creates a premature termination codon (p.Trp4Ter) associated with TEN; a mutant protein is synthesized (likely via PTC-readthrough) and is abnormally localized to the nuclear compartment, potentially leading to a toxic effect rather than the normal secretory pathway.\",\n      \"method\": \"Direct sequencing of GNLY coding region, functional localization assay\",\n      \"journal\": \"Human genetics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — direct mutagenesis with subcellular localization and functional consequence, single lab\",\n      \"pmids\": [\"31642954\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"GNLY+CD8+ T cells predominantly express the pro-inflammatory 15 kDa form of granulysin and are activated by IL-15 in vitro; supernatant from IL-15-stimulated CD8+ T cells induces monocytes to secrete inflammatory factors and disrupts intestinal epithelial cell integrity, effects that can be partially restored by anti-GNLY antibodies.\",\n      \"method\": \"Single-cell RNA sequencing, in vitro IL-15 stimulation, anti-GNLY antibody neutralization assay, monocyte co-culture\",\n      \"journal\": \"Emerging microbes & infections\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — multiple orthogonal methods (scRNA-seq, in vitro stimulation, antibody neutralization) in single study\",\n      \"pmids\": [\"40135938\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"GNLY (granulysin/NKG5) encodes a secreted antimicrobial and cytolytic peptide expressed in NK cells and cytotoxic T lymphocytes upon activation, encoded at chromosome 2p12-q11, produced as alternative splice forms including a pro-inflammatory 15 kDa form; granulysin is released from cytotoxic granules along with granzyme and perforin, can induce inflammatory signaling in monocytes and disrupt epithelial barrier integrity via a paracrine mechanism blocked by anti-GNLY antibodies, and proper N-terminal sequence is required for normal secretory localization since truncation mutants mislocalize to the nucleus.\"\n}\n```","stage2_raw":"```json\n{\n  \"mechanistic_narrative\": \"GNLY encodes granulysin, a secreted cytolytic and pro-inflammatory peptide expressed in natural killer cells and cytotoxic T lymphocytes upon activation, produced as alternative splice forms from a five-exon gene on chromosome 2p12–q11 [PMID:8423048, PMID:2209093]. The 15 kDa pro-inflammatory form of granulysin, released from activated CD8+ T cells stimulated by IL-15, induces monocyte inflammatory cytokine secretion and disrupts intestinal epithelial barrier integrity via a paracellular mechanism that is blocked by anti-GNLY antibodies [PMID:40135938]. The N-terminal signal sequence is required for correct secretory pathway localization, as a premature termination mutation (p.Trp4Ter) yields a protein that mislocalizes to the nucleus with potential toxic consequences, linking GNLY to toxic epidermal necrolysis (TEN) [PMID:31642954].\",\n  \"teleology\": [\n    {\n      \"year\": 1990,\n      \"claim\": \"Chromosomal mapping established that GNLY resides at 2p12–q11, placing it in a genomic context distinct from other cytolytic effector gene clusters and enabling subsequent cloning efforts.\",\n      \"evidence\": \"In situ hybridization on human metaphase chromosomes\",\n      \"pmids\": [\"2209093\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Gene structure and transcript diversity were not yet characterized\",\n        \"No functional data for the encoded protein\"\n      ]\n    },\n    {\n      \"year\": 1993,\n      \"claim\": \"Cloning and structural analysis revealed that GNLY is a five-exon gene producing alternatively spliced transcripts (NKG5 and clone 519) encoding a secreted protein specifically upregulated in activated NK and T cells, establishing it as an activation-dependent immune effector gene.\",\n      \"evidence\": \"Genomic library screening, sequencing, 5′-flanking region analysis, and cDNA comparison\",\n      \"pmids\": [\"8423048\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Functional roles of the encoded protein (cytolysis, antimicrobial activity) were not tested\",\n        \"Regulation by specific cytokines was not defined\"\n      ]\n    },\n    {\n      \"year\": 1999,\n      \"claim\": \"Identification of granulysin as a mitogenic factor for endothelial cells at sub-nanomolar concentrations raised the possibility that granulysin has paracrine signaling roles beyond direct cytolysis.\",\n      \"evidence\": \"Heparin-sepharose fractionation of decidual extract, endothelial proliferation assay, protein sequencing\",\n      \"pmids\": [\"10584980\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\n        \"Single functional assay without receptor identification or signaling pathway dissection\",\n        \"No in vivo validation of angiogenic activity\",\n        \"Not independently replicated\"\n      ]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"A premature termination mutation (p.Trp4Ter) demonstrated that the N-terminal signal peptide is essential for secretory pathway targeting; the truncated protein mislocalizes to the nucleus and is associated with toxic epidermal necrolysis, linking GNLY loss-of-function to severe adverse drug reactions.\",\n      \"evidence\": \"Direct sequencing of GNLY coding region and subcellular localization assay of mutant protein\",\n      \"pmids\": [\"31642954\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Mechanism of nuclear toxicity not elucidated\",\n        \"Single patient/family study — broader population confirmation needed\",\n        \"Whether PTC-readthrough product retains partial cytolytic activity is unknown\"\n      ]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Mechanistic dissection showed that IL-15-activated CD8+ T cells produce the 15 kDa pro-inflammatory granulysin isoform, which acts in a paracrine manner to activate monocyte inflammatory responses and compromise intestinal epithelial barrier integrity — effects reversible by anti-GNLY antibodies — establishing granulysin as a mediator of tissue-damaging inflammation.\",\n      \"evidence\": \"Single-cell RNA-seq, IL-15 in vitro stimulation, monocyte co-culture, anti-GNLY antibody neutralization, epithelial barrier assays\",\n      \"pmids\": [\"40135938\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Receptor or signaling pathway on monocytes and epithelial cells not identified\",\n        \"In vivo relevance in gut inflammation not demonstrated\",\n        \"Relative contributions of 9 kDa vs 15 kDa isoforms to barrier disruption not resolved\"\n      ]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"The receptor(s) through which granulysin signals on monocytes and epithelial cells, the structural basis of its membrane-disrupting activity against host versus microbial membranes, and the in vivo contribution of each splice isoform to immune defense versus tissue damage remain undefined.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"gaps\": [\n        \"No receptor identified for granulysin on target host cells\",\n        \"No structural model explaining selectivity for microbial vs host membranes\",\n        \"In vivo isoform-specific knockout studies lacking\"\n      ]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0048018\", \"supporting_discovery_ids\": [4]},\n      {\"term_id\": \"GO:0090729\", \"supporting_discovery_ids\": [4]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0031410\", \"supporting_discovery_ids\": [0, 4]},\n      {\"term_id\": \"GO:0005576\", \"supporting_discovery_ids\": [0, 4]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [0, 4]}\n    ],\n    \"complexes\": [],\n    \"partners\": [],\n    \"other_free_text\": []\n  }\n}\n```"}