{"gene":"MAP3K14","run_date":"2026-06-14T21:11:36+00:00","timeline":{"discoveries":[{"year":2005,"finding":"RSV infection induces NIK (MAP3K14) kinase activity within 6 h of adsorption, leading to NIK complex formation with IKKα and NF-κB2 (p100/p52). NIK mediates processing of the 100-kDa NF-κB2 precursor into its 52-kDa DNA-binding isoform. Notably, NIK transiently translocates into the nucleus complexed with 52-kDa NF-κB2. siRNA-mediated NIK knockdown blocked RSV-inducible NF-κB2 p52 processing and interfered with early activation of a subset of NF-κB-dependent genes, establishing NIK as an upstream activator of the noncanonical NF-κB pathway in RSV infection.","method":"Immunoprecipitation kinase assay, coimmunoprecipitation, Western immunoblot, microaffinity DNA pull-down, siRNA knockdown","journal":"Journal of virology","confidence":"High","confidence_rationale":"Tier 2 / Moderate — reciprocal Co-IP, kinase activity assay, siRNA loss-of-function with defined molecular phenotype, multiple orthogonal methods in a single study","pmids":["15994789"],"is_preprint":false},{"year":2016,"finding":"MAP3K14 (NIK) is required for RANKL-induced M cell differentiation in intestinal enteroids. Enteroids from Map3k14 null-mutant mice failed to induce M cell-associated genes (Spib, Ccl9, Tnfaip2, Anxa5, Marcksl1, Gp2) upon RANKL stimulation, placing MAP3K14-dependent noncanonical NF-κB signaling as an obligate pathway downstream of RANKL receptor activation for M cell differentiation.","method":"Genetic loss-of-function (Map3k14 null mice), 3D enteroid culture, gene expression analysis","journal":"American journal of physiology. Cell physiology","confidence":"High","confidence_rationale":"Tier 2 / Moderate — clean genetic KO with defined cellular differentiation phenotype and multiple molecular readouts in a physiologically relevant ex vivo model","pmids":["27413168"],"is_preprint":false},{"year":2016,"finding":"MAP3K14 promotes acute kidney injury (AKI) through tubular cell inflammation and apoptosis via the noncanonical NF-κB pathway. MAP3K14 activity-deficient (aly/aly) mice showed less kidney dysfunction, inflammation, and apoptosis in folic acid- and cisplatin-induced AKI models. Bone marrow exchange experiments showed the protective effect was independent of immune cell MAP3K14, implicating tubular cell MAP3K14. In cultured tubular cells, MAP3K14 siRNA decreased inflammation and cell death, and the chemokines MCP-1, RANTES, and CXCL10 were identified as MAP3K14 targets.","method":"MAP3K14 activity-deficient (aly/aly) mouse model, bone marrow transplantation, siRNA knockdown in tubular cell culture, DNA binding assay (RelB/NF-κB2 nuclear fractions), immunohistochemistry","journal":"Journal of the American Society of Nephrology : JASN","confidence":"High","confidence_rationale":"Tier 2 / Moderate — genetic loss-of-function in vivo, bone marrow exchange epistasis, siRNA in vitro, multiple orthogonal readouts across two AKI models","pmids":["27620989"],"is_preprint":false},{"year":2019,"finding":"MAP3K14 (NIK) is a direct target of ssc-novel-miR-106-5p in porcine endometrial epithelial cells. miR-106-5p mimic downregulated MAP3K14 mRNA and protein, inhibited NF-κB phosphorylation, and decreased IL-1 and TNF-α secretion, while miR-106-5p inhibitor had the opposite effect, placing MAP3K14 as a positive upstream regulator of NF-κB activation and proinflammatory cytokine production in endometrial cells.","method":"Dual-luciferase reporter assay (direct miRNA target validation), miRNA mimic/inhibitor transfection, Western blot, cytokine measurement","journal":"Reproduction, fertility, and development","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — luciferase reporter validates direct targeting, functional rescue with mimic/inhibitor, single lab","pmids":["31242957"],"is_preprint":false},{"year":2023,"finding":"MAP3K14 is a direct target of gga-miR-20a-5p in chicken macrophages. Luciferase reporter assay confirmed direct binding; exosome-mediated delivery of gga-miR-20a-5p repressed MAP3K14 expression and downstream proinflammatory cytokine (IFNG, IL1B, TNFA) production, consistent with MAP3K14's role upstream of NF-κB/MAPK-driven inflammatory signaling.","method":"Luciferase reporter assay, exosome-mediated miRNA delivery, qRT-PCR","journal":"Animal bioscience","confidence":"Low","confidence_rationale":"Tier 3 / Weak — luciferase reporter confirms targeting, but functional mechanistic detail on MAP3K14 pathway is indirect; single lab, single method per readout","pmids":["36634655"],"is_preprint":false},{"year":2019,"finding":"Computational structural analysis revealed that selectivity of type I 1/2 kinase inhibitors between MAP3K14 (NIK) and PAK4 is determined by differences in protein-ligand interactions, positions and conformations of key active-site residues, and ligand flexibilities, reflecting the distinct binding pocket architecture of MAP3K14.","method":"Molecular docking, QM/MM simulation, molecular dynamics simulation, DFT calculation","journal":"Physical chemistry chemical physics : PCCP","confidence":"Low","confidence_rationale":"Tier 4 / Weak — computational/modeling study only, no experimental mutagenesis or biochemical validation of MAP3K14 binding mode","pmids":["31657381"],"is_preprint":false}],"current_model":"MAP3K14 (NIK) is a serine/threonine kinase that serves as the master upstream activator of the noncanonical NF-κB pathway: upon activation (e.g., by RSV infection or RANKL), NIK forms complexes with IKKα and NF-κB2 (p100), phosphorylates and promotes proteolytic processing of p100 to the p52 DNA-binding isoform, and can translocate to the nucleus complexed with p52; genetic loss-of-function studies demonstrate that MAP3K14 is required for RANKL-driven intestinal M cell differentiation and promotes acute kidney injury through tubular cell inflammation and apoptosis via noncanonical NF-κB–dependent chemokine production."},"narrative":{"mechanistic_narrative":"MAP3K14 (NIK) is a serine/threonine kinase that functions as the upstream activator of the noncanonical NF-κB pathway [PMID:15994789]. Upon activation by stimuli such as RSV infection, NIK forms complexes with IKKα and NF-κB2 (p100), drives proteolytic processing of the 100-kDa p100 precursor into the 52-kDa p52 DNA-binding isoform, and transiently translocates into the nucleus complexed with p52, thereby driving a subset of NF-κB-dependent genes [PMID:15994789]. This kinase-driven noncanonical signaling axis is obligate for RANKL-induced intestinal M cell differentiation, where loss of MAP3K14 abolishes induction of M cell genes including Spib, Ccl9, and Gp2 [PMID:27413168]. In the kidney, tubular-cell MAP3K14 promotes acute kidney injury by driving inflammation and apoptosis through noncanonical NF-κB–dependent production of the chemokines MCP-1, RANTES, and CXCL10 [PMID:27620989]. Across endometrial epithelial and macrophage systems, MAP3K14 acts as a positive upstream regulator of NF-κB activation and proinflammatory cytokine output and is post-transcriptionally repressed by specific microRNAs [PMID:31242957, PMID:36634655].","teleology":[{"year":2005,"claim":"Established MAP3K14/NIK as an upstream activator of the noncanonical NF-κB pathway by showing it physically assembles with IKKα and p100 and drives p100-to-p52 processing during RSV infection.","evidence":"IP kinase assay, reciprocal Co-IP, DNA pull-down, and siRNA knockdown in RSV-infected cells","pmids":["15994789"],"confidence":"High","gaps":["Direct phosphorylation site(s) on IKKα or p100 by NIK not mapped in this study","Mechanism of NIK nuclear translocation with p52 not defined","Activating signal upstream of NIK in RSV infection not resolved"]},{"year":2016,"claim":"Demonstrated that MAP3K14 is an obligate node downstream of the RANKL receptor for a differentiation program, extending its role beyond inflammation to cell-fate specification.","evidence":"Genetic loss-of-function in Map3k14-null mouse enteroids with M cell gene-expression readouts","pmids":["27413168"],"confidence":"High","gaps":["Direct transcriptional targets bridging NIK and M cell genes not identified","Whether p52/RelB directly binds M cell gene promoters not shown"]},{"year":2016,"claim":"Localized MAP3K14's pathogenic role in acute kidney injury to tubular cells and identified specific chemokine effectors, distinguishing tissue-intrinsic from immune-cell contributions.","evidence":"aly/aly kinase-deficient mice, bone marrow transplantation epistasis, and tubular-cell siRNA across two AKI models","pmids":["27620989"],"confidence":"High","gaps":["Direct kinase substrates linking NIK to chemokine induction not defined","Whether MCP-1/RANTES/CXCL10 are direct p52/RelB transcriptional targets not established"]},{"year":2019,"claim":"Positioned MAP3K14 under post-transcriptional control, showing miRNA-mediated repression tunes NF-κB activation and cytokine output.","evidence":"Dual-luciferase reporter validation of direct miR-106-5p targeting plus mimic/inhibitor functional assays in porcine endometrial epithelial cells","pmids":["31242957"],"confidence":"Medium","gaps":["Single-lab finding in a non-human system","Canonical vs noncanonical NF-κB branch not distinguished in the cytokine readout"]},{"year":2023,"claim":"Reinforced miRNA control of MAP3K14 in a second species and cell type, with exosomal delivery repressing the kinase and downstream cytokines.","evidence":"Luciferase reporter and exosome-mediated miR-20a-5p delivery in chicken macrophages with qRT-PCR cytokine readouts","pmids":["36634655"],"confidence":"Low","gaps":["Mechanistic link to MAP3K14 pathway is indirect","Single lab, single method per readout"]},{"year":2019,"claim":"Characterized the determinants of small-molecule inhibitor selectivity at the MAP3K14 active site, informing kinase-targeted design.","evidence":"Computational docking, QM/MM, molecular dynamics, and DFT comparison of NIK versus PAK4 binding pockets","pmids":["31657381"],"confidence":"Low","gaps":["No experimental mutagenesis or biochemical validation of the predicted binding mode","No co-crystal structure reported in the timeline"]},{"year":null,"claim":"The direct phosphorylation substrates of NIK and the structural basis of its assembly with IKKα and p100 remain undefined in the available corpus.","evidence":"","pmids":[],"confidence":"Low","gaps":["No mapped NIK phosphosites on IKKα or p100","No experimental structure of the NIK–IKKα–p100 complex","Regulation of NIK protein stability/turnover not characterized here"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0140096","term_label":"catalytic activity, acting on a protein","supporting_discovery_ids":[0]},{"term_id":"GO:0016740","term_label":"transferase activity","supporting_discovery_ids":[0]}],"localization":[{"term_id":"GO:0005829","term_label":"cytosol","supporting_discovery_ids":[0]},{"term_id":"GO:0005634","term_label":"nucleus","supporting_discovery_ids":[0]}],"pathway":[{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[0,2]},{"term_id":"R-HSA-168256","term_label":"Immune System","supporting_discovery_ids":[2,4]},{"term_id":"R-HSA-1266738","term_label":"Developmental Biology","supporting_discovery_ids":[1]}],"complexes":[],"partners":["IKKA","NFKB2"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q99558","full_name":"Mitogen-activated protein kinase kinase kinase 14","aliases":["NF-kappa-beta-inducing kinase","HsNIK","Serine/threonine-protein kinase NIK"],"length_aa":947,"mass_kda":104.0,"function":"Lymphotoxin beta-activated kinase which seems to be exclusively involved in the activation of NF-kappa-B and its transcriptional activity. Phosphorylates CHUK/IKKA, thereby promoting proteolytic processing of NFKB2/P100, which leads to NF-kappa-B activation via the non-canonical pathway (PubMed:25406581, PubMed:29230214). Has an essential role in the non-canonical NF-kappa-B signaling that regulates genes encoding molecules involved in B-cell survival, lymphoid organogenesis, and immune response (PubMed:25406581). Could act in a receptor-selective manner","subcellular_location":"Cytoplasm","url":"https://www.uniprot.org/uniprotkb/Q99558/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/MAP3K14","classification":"Not Classified","n_dependent_lines":4,"n_total_lines":74,"dependency_fraction":0.05405405405405406},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/MAP3K14","total_profiled":1310},"omim":[{"mim_id":"620449","title":"IMMUNODEFICIENCY 112; IMD112","url":"https://www.omim.org/entry/620449"},{"mim_id":"619289","title":"ZINC FINGER PROTEIN 91, ATYPICAL E3 UBIQUITIN LIGASE; ZFP91","url":"https://www.omim.org/entry/619289"},{"mim_id":"618428","title":"POPOV-CHANG SYNDROME; POPCHAS","url":"https://www.omim.org/entry/618428"},{"mim_id":"615577","title":"IMMUNODEFICIENCY, COMMON VARIABLE, 10; CVID10","url":"https://www.omim.org/entry/615577"},{"mim_id":"614061","title":"OLFACTOMEDIN 4; OLFM4","url":"https://www.omim.org/entry/614061"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Supported","locations":[{"location":"Vesicles","reliability":"Supported"},{"location":"Cytosol","reliability":"Supported"},{"location":"Nucleoplasm","reliability":"Additional"},{"location":"Nucleoli fibrillar center","reliability":"Additional"}],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in all","driving_tissues":[],"url":"https://www.proteinatlas.org/search/MAP3K14"},"hgnc":{"alias_symbol":["NIK","HSNIK","FTDCR1B","HS"],"prev_symbol":[]},"alphafold":{"accession":"Q99558","domains":[{"cath_id":"3.30.200.20","chopping":"334-367_374-470","consensus_level":"high","plddt":91.5405,"start":334,"end":470},{"cath_id":"1.10.510.10","chopping":"478-659","consensus_level":"high","plddt":91.8634,"start":478,"end":659},{"cath_id":"3.10.20.90","chopping":"860-947","consensus_level":"medium","plddt":87.025,"start":860,"end":947}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q99558","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q99558-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q99558-F1-predicted_aligned_error_v6.png","plddt_mean":60.75},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=MAP3K14","jax_strain_url":"https://www.jax.org/strain/search?query=MAP3K14"},"sequence":{"accession":"Q99558","fasta_url":"https://rest.uniprot.org/uniprotkb/Q99558.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q99558/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q99558"}},"corpus_meta":[{"pmid":"15994789","id":"PMC_15994789","title":"Respiratory syncytial virus influences NF-kappaB-dependent gene expression through a novel pathway involving MAP3K14/NIK expression and nuclear complex formation with NF-kappaB2.","date":"2005","source":"Journal of virology","url":"https://pubmed.ncbi.nlm.nih.gov/15994789","citation_count":61,"is_preprint":false},{"pmid":"25483963","id":"PMC_25483963","title":"Autophagy-dependent PELI3 degradation inhibits proinflammatory IL1B expression.","date":"2014","source":"Autophagy","url":"https://pubmed.ncbi.nlm.nih.gov/25483963","citation_count":59,"is_preprint":false},{"pmid":"27413168","id":"PMC_27413168","title":"TNF-α augments RANKL-dependent intestinal M cell differentiation in enteroid cultures.","date":"2016","source":"American journal of physiology. Cell physiology","url":"https://pubmed.ncbi.nlm.nih.gov/27413168","citation_count":59,"is_preprint":false},{"pmid":"21531130","id":"PMC_21531130","title":"Differential gene expression signatures between colorectal cancers with and without KRAS mutations: crosstalk between the KRAS pathway and other signalling pathways.","date":"2011","source":"European journal of cancer (Oxford, England : 1990)","url":"https://pubmed.ncbi.nlm.nih.gov/21531130","citation_count":53,"is_preprint":false},{"pmid":"27620989","id":"PMC_27620989","title":"Mitogen-Activated Protein Kinase 14 Promotes AKI.","date":"2016","source":"Journal of the American Society of Nephrology : JASN","url":"https://pubmed.ncbi.nlm.nih.gov/27620989","citation_count":37,"is_preprint":false},{"pmid":"37455213","id":"PMC_37455213","title":"Comprehensive Analyses Revealed Eight Immune Related Signatures Correlated With Aberrant Methylations as Prognosis and Diagnosis Biomarkers for Kidney Renal Papillary Cell Carcinoma.","date":"2023","source":"Clinical genitourinary cancer","url":"https://pubmed.ncbi.nlm.nih.gov/37455213","citation_count":25,"is_preprint":false},{"pmid":"31657381","id":"PMC_31657381","title":"Molecular dynamics simulation and QM/MM calculation reveal the selectivity mechanism of type I 1/2 kinase inhibitors: the effect of intramolecular H-bonds and conformational restriction for improved selectivity.","date":"2019","source":"Physical chemistry chemical physics : PCCP","url":"https://pubmed.ncbi.nlm.nih.gov/31657381","citation_count":23,"is_preprint":false},{"pmid":"31922225","id":"PMC_31922225","title":"Silencing CDC25A inhibits the proliferation of liver cancer cells by downregulating IL‑6 in vitro and in vivo.","date":"2020","source":"International journal of molecular medicine","url":"https://pubmed.ncbi.nlm.nih.gov/31922225","citation_count":20,"is_preprint":false},{"pmid":"36634655","id":"PMC_36634655","title":"Exosome-mediated delivery of gga-miR-20a-5p regulates immune response of chicken macrophages by targeting IFNGR2, MAPK1, MAP3K5, and MAP3K14.","date":"2023","source":"Animal bioscience","url":"https://pubmed.ncbi.nlm.nih.gov/36634655","citation_count":5,"is_preprint":false},{"pmid":"31242957","id":"PMC_31242957","title":"Ssc-novel-miR-106-5p reduces lipopolysaccharide-induced inflammatory response in porcine endometrial epithelial cells by inhibiting the expression of the target gene mitogen-activated protein kinase kinase kinase 14 (MAP3K14).","date":"2019","source":"Reproduction, fertility, and development","url":"https://pubmed.ncbi.nlm.nih.gov/31242957","citation_count":4,"is_preprint":false},{"pmid":"40014404","id":"PMC_40014404","title":"The Role of microRNAs in Lidocaine-Induced Spinal Cord Neurotoxicity: An Exploration Based on Bioinformatics Analysis.","date":"2025","source":"DNA and cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/40014404","citation_count":0,"is_preprint":false},{"pmid":"42152489","id":"PMC_42152489","title":"Influenza a virus NS2 suppresses NFKB/NF-κB signaling to facilitate viral replication by mediating the autophagic-degradation of IKBKG/NEMO.","date":"2026","source":"Autophagy","url":"https://pubmed.ncbi.nlm.nih.gov/42152489","citation_count":0,"is_preprint":false}],"cost":{"stage1":{"sonnet_model":"claude-sonnet-4-6","sonnet_input_tokens":10233,"sonnet_output_tokens":1922,"sonnet_usd":0.0595,"sonnet_elapsed_seconds":37.2,"stage1_stop_reason":"end_turn"},"stage2":{"opus_model":"claude-opus-4-8","opus_input_tokens":8870,"opus_output_tokens":2329,"opus_usd":0.1026,"opus_elapsed_seconds":25.0,"stage2_stop_reason":"end_turn"},"total_usd":0.1621},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2005,\n      \"finding\": \"RSV infection induces NIK (MAP3K14) kinase activity within 6 h of adsorption, leading to NIK complex formation with IKKα and NF-κB2 (p100/p52). NIK mediates processing of the 100-kDa NF-κB2 precursor into its 52-kDa DNA-binding isoform. Notably, NIK transiently translocates into the nucleus complexed with 52-kDa NF-κB2. siRNA-mediated NIK knockdown blocked RSV-inducible NF-κB2 p52 processing and interfered with early activation of a subset of NF-κB-dependent genes, establishing NIK as an upstream activator of the noncanonical NF-κB pathway in RSV infection.\",\n      \"method\": \"Immunoprecipitation kinase assay, coimmunoprecipitation, Western immunoblot, microaffinity DNA pull-down, siRNA knockdown\",\n      \"journal\": \"Journal of virology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reciprocal Co-IP, kinase activity assay, siRNA loss-of-function with defined molecular phenotype, multiple orthogonal methods in a single study\",\n      \"pmids\": [\"15994789\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"MAP3K14 (NIK) is required for RANKL-induced M cell differentiation in intestinal enteroids. Enteroids from Map3k14 null-mutant mice failed to induce M cell-associated genes (Spib, Ccl9, Tnfaip2, Anxa5, Marcksl1, Gp2) upon RANKL stimulation, placing MAP3K14-dependent noncanonical NF-κB signaling as an obligate pathway downstream of RANKL receptor activation for M cell differentiation.\",\n      \"method\": \"Genetic loss-of-function (Map3k14 null mice), 3D enteroid culture, gene expression analysis\",\n      \"journal\": \"American journal of physiology. Cell physiology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — clean genetic KO with defined cellular differentiation phenotype and multiple molecular readouts in a physiologically relevant ex vivo model\",\n      \"pmids\": [\"27413168\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"MAP3K14 promotes acute kidney injury (AKI) through tubular cell inflammation and apoptosis via the noncanonical NF-κB pathway. MAP3K14 activity-deficient (aly/aly) mice showed less kidney dysfunction, inflammation, and apoptosis in folic acid- and cisplatin-induced AKI models. Bone marrow exchange experiments showed the protective effect was independent of immune cell MAP3K14, implicating tubular cell MAP3K14. In cultured tubular cells, MAP3K14 siRNA decreased inflammation and cell death, and the chemokines MCP-1, RANTES, and CXCL10 were identified as MAP3K14 targets.\",\n      \"method\": \"MAP3K14 activity-deficient (aly/aly) mouse model, bone marrow transplantation, siRNA knockdown in tubular cell culture, DNA binding assay (RelB/NF-κB2 nuclear fractions), immunohistochemistry\",\n      \"journal\": \"Journal of the American Society of Nephrology : JASN\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic loss-of-function in vivo, bone marrow exchange epistasis, siRNA in vitro, multiple orthogonal readouts across two AKI models\",\n      \"pmids\": [\"27620989\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"MAP3K14 (NIK) is a direct target of ssc-novel-miR-106-5p in porcine endometrial epithelial cells. miR-106-5p mimic downregulated MAP3K14 mRNA and protein, inhibited NF-κB phosphorylation, and decreased IL-1 and TNF-α secretion, while miR-106-5p inhibitor had the opposite effect, placing MAP3K14 as a positive upstream regulator of NF-κB activation and proinflammatory cytokine production in endometrial cells.\",\n      \"method\": \"Dual-luciferase reporter assay (direct miRNA target validation), miRNA mimic/inhibitor transfection, Western blot, cytokine measurement\",\n      \"journal\": \"Reproduction, fertility, and development\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — luciferase reporter validates direct targeting, functional rescue with mimic/inhibitor, single lab\",\n      \"pmids\": [\"31242957\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"MAP3K14 is a direct target of gga-miR-20a-5p in chicken macrophages. Luciferase reporter assay confirmed direct binding; exosome-mediated delivery of gga-miR-20a-5p repressed MAP3K14 expression and downstream proinflammatory cytokine (IFNG, IL1B, TNFA) production, consistent with MAP3K14's role upstream of NF-κB/MAPK-driven inflammatory signaling.\",\n      \"method\": \"Luciferase reporter assay, exosome-mediated miRNA delivery, qRT-PCR\",\n      \"journal\": \"Animal bioscience\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — luciferase reporter confirms targeting, but functional mechanistic detail on MAP3K14 pathway is indirect; single lab, single method per readout\",\n      \"pmids\": [\"36634655\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"Computational structural analysis revealed that selectivity of type I 1/2 kinase inhibitors between MAP3K14 (NIK) and PAK4 is determined by differences in protein-ligand interactions, positions and conformations of key active-site residues, and ligand flexibilities, reflecting the distinct binding pocket architecture of MAP3K14.\",\n      \"method\": \"Molecular docking, QM/MM simulation, molecular dynamics simulation, DFT calculation\",\n      \"journal\": \"Physical chemistry chemical physics : PCCP\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 4 / Weak — computational/modeling study only, no experimental mutagenesis or biochemical validation of MAP3K14 binding mode\",\n      \"pmids\": [\"31657381\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"MAP3K14 (NIK) is a serine/threonine kinase that serves as the master upstream activator of the noncanonical NF-κB pathway: upon activation (e.g., by RSV infection or RANKL), NIK forms complexes with IKKα and NF-κB2 (p100), phosphorylates and promotes proteolytic processing of p100 to the p52 DNA-binding isoform, and can translocate to the nucleus complexed with p52; genetic loss-of-function studies demonstrate that MAP3K14 is required for RANKL-driven intestinal M cell differentiation and promotes acute kidney injury through tubular cell inflammation and apoptosis via noncanonical NF-κB–dependent chemokine production.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"MAP3K14 (NIK) is a serine/threonine kinase that functions as the upstream activator of the noncanonical NF-κB pathway [#0]. Upon activation by stimuli such as RSV infection, NIK forms complexes with IKKα and NF-κB2 (p100), drives proteolytic processing of the 100-kDa p100 precursor into the 52-kDa p52 DNA-binding isoform, and transiently translocates into the nucleus complexed with p52, thereby driving a subset of NF-κB-dependent genes [#0]. This kinase-driven noncanonical signaling axis is obligate for RANKL-induced intestinal M cell differentiation, where loss of MAP3K14 abolishes induction of M cell genes including Spib, Ccl9, and Gp2 [#1]. In the kidney, tubular-cell MAP3K14 promotes acute kidney injury by driving inflammation and apoptosis through noncanonical NF-κB–dependent production of the chemokines MCP-1, RANTES, and CXCL10 [#2]. Across endometrial epithelial and macrophage systems, MAP3K14 acts as a positive upstream regulator of NF-κB activation and proinflammatory cytokine output and is post-transcriptionally repressed by specific microRNAs [#3, #4].\",\n  \"teleology\": [\n    {\n      \"year\": 2005,\n      \"claim\": \"Established MAP3K14/NIK as an upstream activator of the noncanonical NF-κB pathway by showing it physically assembles with IKKα and p100 and drives p100-to-p52 processing during RSV infection.\",\n      \"evidence\": \"IP kinase assay, reciprocal Co-IP, DNA pull-down, and siRNA knockdown in RSV-infected cells\",\n      \"pmids\": [\"15994789\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Direct phosphorylation site(s) on IKKα or p100 by NIK not mapped in this study\", \"Mechanism of NIK nuclear translocation with p52 not defined\", \"Activating signal upstream of NIK in RSV infection not resolved\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Demonstrated that MAP3K14 is an obligate node downstream of the RANKL receptor for a differentiation program, extending its role beyond inflammation to cell-fate specification.\",\n      \"evidence\": \"Genetic loss-of-function in Map3k14-null mouse enteroids with M cell gene-expression readouts\",\n      \"pmids\": [\"27413168\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Direct transcriptional targets bridging NIK and M cell genes not identified\", \"Whether p52/RelB directly binds M cell gene promoters not shown\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Localized MAP3K14's pathogenic role in acute kidney injury to tubular cells and identified specific chemokine effectors, distinguishing tissue-intrinsic from immune-cell contributions.\",\n      \"evidence\": \"aly/aly kinase-deficient mice, bone marrow transplantation epistasis, and tubular-cell siRNA across two AKI models\",\n      \"pmids\": [\"27620989\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Direct kinase substrates linking NIK to chemokine induction not defined\", \"Whether MCP-1/RANTES/CXCL10 are direct p52/RelB transcriptional targets not established\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Positioned MAP3K14 under post-transcriptional control, showing miRNA-mediated repression tunes NF-κB activation and cytokine output.\",\n      \"evidence\": \"Dual-luciferase reporter validation of direct miR-106-5p targeting plus mimic/inhibitor functional assays in porcine endometrial epithelial cells\",\n      \"pmids\": [\"31242957\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single-lab finding in a non-human system\", \"Canonical vs noncanonical NF-κB branch not distinguished in the cytokine readout\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Reinforced miRNA control of MAP3K14 in a second species and cell type, with exosomal delivery repressing the kinase and downstream cytokines.\",\n      \"evidence\": \"Luciferase reporter and exosome-mediated miR-20a-5p delivery in chicken macrophages with qRT-PCR cytokine readouts\",\n      \"pmids\": [\"36634655\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"Mechanistic link to MAP3K14 pathway is indirect\", \"Single lab, single method per readout\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Characterized the determinants of small-molecule inhibitor selectivity at the MAP3K14 active site, informing kinase-targeted design.\",\n      \"evidence\": \"Computational docking, QM/MM, molecular dynamics, and DFT comparison of NIK versus PAK4 binding pockets\",\n      \"pmids\": [\"31657381\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"No experimental mutagenesis or biochemical validation of the predicted binding mode\", \"No co-crystal structure reported in the timeline\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"The direct phosphorylation substrates of NIK and the structural basis of its assembly with IKKα and p100 remain undefined in the available corpus.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"No mapped NIK phosphosites on IKKα or p100\", \"No experimental structure of the NIK–IKKα–p100 complex\", \"Regulation of NIK protein stability/turnover not characterized here\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0140096\", \"supporting_discovery_ids\": [0]},\n      {\"term_id\": \"GO:0016740\", \"supporting_discovery_ids\": [0]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [0]},\n      {\"term_id\": \"GO:0005634\", \"supporting_discovery_ids\": [0]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [0, 2]},\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [2, 4]},\n      {\"term_id\": \"R-HSA-1266738\", \"supporting_discovery_ids\": [1]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"IKKA\", \"NFKB2\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win"}}