{"gene":"TNFSF12","run_date":"2026-04-28T21:42:59","timeline":{"discoveries":[{"year":2001,"finding":"TWEAK (TNFSF12) binds to TweakR/Fn14, a small TNF receptor family member cloned from HUVEC library, with an affinity constant Kd of 2.3 ± 0.1 nM; the TweakR cytoplasmic domain binds TRAF1, TRAF2, and TRAF3; cross-linking of TweakR induces HUVEC growth; soluble TweakR inhibits endothelial cell migration and corneal angiogenesis in vivo.","method":"Five binding assays including receptor cloning, affinity measurement, Co-IP/pulldown for TRAF binding, HUVEC growth assay, corneal angiogenesis model","journal":"Immunity","confidence":"High","confidence_rationale":"Tier 1–2 — receptor cloning with multiple orthogonal binding assays, TRAF pulldown, and in vivo functional validation in one study","pmids":["11728344"],"is_preprint":false},{"year":2003,"finding":"TWEAK sequentially activates two NF-κB cascades: (1) rapid IκBα phosphorylation generating p50/RelA complexes via canonical IKK complex (NEMO-dependent); (2) prolonged activation via proteasome-mediated NF-κB2/p100 processing generating RelB complexes (NEMO-independent, requiring IKK1 and NF-κB-inducing kinase). Fn14 with mutated TRAF-binding site cannot activate NF-κB; cells deficient in TRAF2 and TRAF5 fail to show p100 processing and IκBα phosphorylation.","method":"Expression cloning, IκBα phosphorylation assays, EMSA, TRAF-binding site mutagenesis, TRAF2/TRAF5 knockout cells, proteasome inhibitor studies","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1–2 — multiple orthogonal methods including mutagenesis, KO cells, and biochemical assays in one study; foundational mechanism paper","pmids":["12840022"],"is_preprint":false},{"year":2002,"finding":"TWEAK-induced cell death occurs through multiple pathways: caspase-8 and caspase-3-dependent apoptosis in HSC3 cells; cathepsin B-dependent necrosis (with lysosomal cathepsin B release to cytosol) in IFN-γ-treated HT-29 cells; and indirect TNF-α-mediated cell death in Kym-1 cells. DR3 is not the relevant receptor as TWEAK-sensitive cell lines do not express DR3 but bind TWEAK via a distinct receptor.","method":"Caspase activity assays, pan-caspase inhibitors, cathepsin B inhibitors, lysosomal fractionation, anti-TNF-α blocking antibodies, DR3 expression analysis by Western blot and RT-PCR","journal":"Journal of immunology","confidence":"High","confidence_rationale":"Tier 1–2 — multiple cell lines with pharmacological inhibitors and mechanistic dissection of distinct death pathways","pmids":["11777967"],"is_preprint":false},{"year":2000,"finding":"TWEAK is expressed on the surface of IFN-γ-stimulated monocytes (but not IFN-α or LPS-stimulated) and mediates cytotoxic killing of tumor cells; anti-TWEAK mAb partially inhibited monocyte cytotoxicity, with near-complete inhibition in combination with anti-TRAIL.","method":"Anti-human TWEAK mAb generation, flow cytometry, cytotoxicity assays with neutralizing antibodies","journal":"The Journal of experimental medicine","confidence":"High","confidence_rationale":"Tier 2 — reciprocal antibody blocking with clear functional readout, replicated finding","pmids":["11067885"],"is_preprint":false},{"year":2000,"finding":"TWEAK and DR3/WSL-1/TRAMP do not interact in in vitro binding assays; TWEAK binds to cells lacking DR3 surface expression and shows biological activity there; cells from DR3 knockout mice retain TWEAK-binding capacity, demonstrating DR3 is not the major TWEAK receptor.","method":"In vitro binding assay, flow cytometry, DR3 knockout mouse cells","journal":"FEBS letters","confidence":"High","confidence_rationale":"Tier 1–2 — direct binding assay and genetic KO cells providing clear negative result for DR3 as TWEAK receptor","pmids":["11094155"],"is_preprint":false},{"year":2002,"finding":"An endogenous hybrid mRNA (TWE-PRIL) encodes a fusion protein composed of TWEAK cytoplasmic and transmembrane domains fused to the APRIL C-terminal receptor-binding domain; TWE-PRIL is membrane-anchored, presents APRIL's receptor-binding domain at the cell surface, and is biologically active (stimulates cycling in T- and B-lymphoma cell lines). Detected in primary human T lymphocytes and monocytic cell lines by RACE, RT-PCR, cDNA library screening, RNase protection assay.","method":"RACE, RT-PCR, cDNA library screening, RNase protection assay, flow cytometry, proliferation assay","journal":"The EMBO journal","confidence":"High","confidence_rationale":"Tier 1–2 — multiple orthogonal methods confirming endogenous expression and biological activity of fusion protein","pmids":["12411489"],"is_preprint":false},{"year":2005,"finding":"TWEAK selectively stimulates liver progenitor (oval) cell proliferation via its receptor Fn14; transgenic TWEAK overexpression in hepatocytes causes oval cell hyperplasia; TWEAK-adenovirus induced oval cell expansion in WT but not Fn14-null mice; DDC-induced oval cell expansion was reduced in Fn14-null mice and by anti-TWEAK mAb; TWEAK stimulated proliferation of oval cell cultures directly.","method":"Transgenic overexpression, adenoviral delivery, Fn14 knockout mice, blocking anti-TWEAK mAb, DDC injury model, cell culture proliferation assays","journal":"The Journal of clinical investigation","confidence":"High","confidence_rationale":"Tier 2 — genetic loss-of-function (Fn14 KO), pharmacological blockade, and gain-of-function (Tg/adenovirus) with concordant phenotypic readouts","pmids":["16110324"],"is_preprint":false},{"year":2006,"finding":"TWEAK binds to mesenchymal progenitor cells (all lineages) and induces NF-κB activation, pro-survival, pro-proliferative and homing receptor gene expression. TWEAK promotes C2C12 myoblast proliferation and inhibits terminal differentiation. Fn14-deficient primary myoblasts show reduced proliferative capacity and altered myotube formation. After cardiotoxin injury, Fn14-deficient mice show reduced inflammatory response and delayed muscle fiber regeneration.","method":"Cell binding assays, NF-κB reporter assays, Fn14 knockout mice, C2C12 and primary myoblast proliferation/differentiation assays, cardiotoxin muscle injury model","journal":"The EMBO journal","confidence":"High","confidence_rationale":"Tier 2 — genetic KO with multiple in vitro and in vivo functional readouts, replicated across cell types","pmids":["17124496"],"is_preprint":false},{"year":2008,"finding":"TWEAK induces NF-κB activation in tubular epithelial cells (MCT line) via IκBα degradation and RelA nuclear translocation, leading to increased MCP-1, RANTES, and IL-6 expression. In vivo systemic TWEAK administration induces renal NF-κB activation, chemokine expression and interstitial inflammation; neutralization of TWEAK with blocking antibody reduces tubular chemokine expression and macrophage infiltration after folic acid-induced AKI. Parthenolide (IκBα degradation inhibitor) blocked all TWEAK-induced effects.","method":"In vitro NF-κB activation assays, Western blot for IκBα, nuclear translocation assays, ELISA, in vivo TWEAK administration in mice, anti-TWEAK mAb neutralization, folic acid AKI model","journal":"Journal of the American Society of Nephrology","confidence":"High","confidence_rationale":"Tier 2 — mechanistic pathway dissection with pharmacological inhibition and in vivo neutralizing antibody validation","pmids":["18235096"],"is_preprint":false},{"year":2010,"finding":"Full-length human TWEAK is processed intracellularly by the serine protease furin at amino acid residues 90–93 (the predominant furin recognition site). Full-length, membrane-anchored TWEAK can bind Fn14 on neighboring cells (juxtacrine signaling) and activate the NF-κB signaling pathway.","method":"Furin inhibitor studies, site-directed mutagenesis of furin cleavage site, co-culture assays, NF-κB reporter assays","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1 — mutagenesis of active site combined with functional NF-κB activation assay, mechanistically defining furin cleavage and juxtacrine activity","pmids":["20385556"],"is_preprint":false},{"year":2006,"finding":"TWEAK/Fn14 interaction in osteoblastic MC3T3-E1 cells induces RANTES via PI3K-Akt (not NF-κB); inhibits BMP-2-induced osteoblast differentiation markers via MAPK-ERK; and upregulates RANKL expression via MAPK-ERK pathway. All effects abolished by Fn14-Fc chimera decoy receptor.","method":"PI3K inhibitors, ERK inhibitors, MAPK pathway analysis, BMP-2 differentiation assays, RANKL expression measurement, Fn14-Fc neutralization","journal":"Arthritis research & therapy","confidence":"Medium","confidence_rationale":"Tier 2 — pathway dissection with pharmacological inhibitors and decoy receptor blockade in single study","pmids":["16945157"],"is_preprint":false},{"year":2011,"finding":"TWEAK and TNFα reduce renal Klotho expression through NF-κB-dependent mechanism. TWEAK promotes RelA binding to the Klotho promoter and induces histone deacetylation of the Klotho promoter. NF-κB inhibitor parthenolide and histone deacetylase inhibitor both prevent TWEAK-induced Klotho downregulation. Confirmed in vivo: TWEAK blockade or genetic absence abrogated injury-related decrease in renal Klotho.","method":"siRNA knockdown of IκBα, parthenolide NF-κB inhibition, HDAC inhibitor, chromatin immunoprecipitation (ChIP), TWEAK knockout mice, anti-TWEAK antibody in folic acid AKI model","journal":"Journal of the American Society of Nephrology","confidence":"High","confidence_rationale":"Tier 1–2 — ChIP showing direct RelA promoter binding, multiple pharmacological inhibitors and genetic KO with concordant results","pmids":["21719790"],"is_preprint":false},{"year":2012,"finding":"TWEAK activates the noncanonical NF-κB pathway (through p100 processing, RelB, IKKα, and NIK) to promote myoblast fusion into multinucleated myotubes. Loss of cIAP1 leads to constitutive noncanonical NF-κB activation and increased myotube nuclei. Low-dose TWEAK, which preferentially activates the noncanonical pathway, increases myoblast fusion without causing atrophy.","method":"cIAP1 knockout, siRNA knockdown of p100/RelB/IKKα/NIK/TRAF3, p52/RelB overexpression, NF-κB reporter assays, myotube fusion quantification","journal":"Science signaling","confidence":"High","confidence_rationale":"Tier 1–2 — multiple genetic perturbations (KO, siRNA, overexpression) with consistent phenotypic readouts establishing noncanonical NF-κB as the pathway for TWEAK-induced myoblast fusion","pmids":["23074266"],"is_preprint":false},{"year":2013,"finding":"TWEAK transactivates EGFR in renal tubular epithelial cells via Fn14-dependent ADAM17 activation and subsequent release of EGFR ligands HB-EGF and TGFα. TWEAK-induced proinflammatory factor expression and ERK activation require EGFR signaling (blocked by erlotinib or ADAM17 inhibitor), while TWEAK-mediated NF-κB activation is EGFR-independent.","method":"EGFR phosphorylation assays, ADAM17 inhibitor (WTACE-2), EGFR kinase inhibitor (erlotinib), in vivo TWEAK administration in mice, HB-EGF/TGFα measurement","journal":"The Journal of pathology","confidence":"High","confidence_rationale":"Tier 1–2 — pharmacological dissection of ADAM17-EGFR axis and in vivo validation, mechanistically separating EGFR-dependent ERK from EGFR-independent NF-κB pathways","pmids":["24037740"],"is_preprint":false},{"year":2015,"finding":"CD163 functions as a decoy receptor for soluble TWEAK during ischemia; soluble CD163 sequesters TWEAK and inhibits TWEAK-induced canonical NF-κB and Notch signaling required for myogenic progenitor cell proliferation. CD163-knockout mice have transiently elevated TWEAK levels, stimulating muscle satellite cell proliferation and tissue regeneration.","method":"CD163 knockout mice, soluble TWEAK binding assays, NF-κB and Notch signaling assays, ischemic injury model, macrophage internalization experiments","journal":"Nature communications","confidence":"High","confidence_rationale":"Tier 2 — genetic KO with mechanistic signaling readouts and in vivo ischemia model validation","pmids":["26242746"],"is_preprint":false},{"year":2009,"finding":"CD163-expressing macrophages can bind and internalize TWEAK protein (demonstrated in vitro with exogenously added TWEAK from supernatants), suggesting CD163 is a scavenger receptor for TWEAK.","method":"In vitro TWEAK binding and internalization assay with CD163-expressing macrophages","journal":"Atherosclerosis","confidence":"Medium","confidence_rationale":"Tier 3 — single-method in vitro binding/internalization experiment","pmids":["19473660"],"is_preprint":false},{"year":2013,"finding":"TWEAK/Fn14 signaling regulates skeletal muscle PGC-1α levels: TWEAK reduces PGC-1α and mitochondrial content (~50%) in skeletal muscle. PGC-1α levels are significantly increased in TWEAK-KO and Fn14-KO mice upon denervation. Transgenic PGC-1α overexpression inhibits muscle wasting in TWEAK-Tg mice, inhibits TWEAK-induced NF-κB activation (~50%), reduces atrogene (MAFbx, MuRF1) expression (~90%), and prevents Fn14 induction in denervated muscle.","method":"TWEAK-KO and Fn14-KO mice, TWEAK-Tg mice, PGC-1α Tg overexpression, denervation model, NF-κB activation assays, atrogene expression measurement, mitochondrial content quantification","journal":"FASEB journal","confidence":"High","confidence_rationale":"Tier 2 — multiple genetic models (KO and Tg) with consistent mechanistic readouts","pmids":["24327607"],"is_preprint":false},{"year":2016,"finding":"TWEAK decreases PGC-1α expression in tubular cells via NF-κB-dependent histone H3 deacetylation at the PGC-1α promoter, leading to decreased expression of PGC-1α target genes and reduced mitochondrial membrane potential. HDAC inhibitors and NF-κB inhibitors both prevent TWEAK-induced PGC-1α downregulation. Adenoviral PGC-1α overexpression rescues TWEAK-induced mitochondrial dysfunction.","method":"Chromatin immunoprecipitation (histone deacetylation at PGC-1α promoter), NF-κB inhibitors, HDAC inhibitors, adenoviral PGC-1α overexpression, mitochondrial membrane potential assays, neutralizing anti-TWEAK antibodies in folic acid AKI","journal":"Kidney international","confidence":"High","confidence_rationale":"Tier 1–2 — ChIP for mechanistic epigenetic regulation, multiple pharmacological inhibitors, in vivo neutralization, and rescue experiment","pmids":["26535995"],"is_preprint":false},{"year":2018,"finding":"TWEAK/Fn14 activation drives a second wave of tubular cell death during AKI through necroptosis. In cultured tubular cells, TWEAK induces apoptosis in a proinflammatory environment; caspase inhibition switches this to necroptosis. In folic acid AKI mice, Fn14 ablation and RIPK1 inhibition (Nec-1) or RIPK3/MLKL deficiency protect against a late (72–96 h) wave of cell death and kidney dysfunction, but not early (48 h) cell death.","method":"Necrostatin-1 (RIPK1 inhibitor), genetic RIPK3 deficiency, MLKL deficiency, Fn14 genetic ablation, caspase inhibitors, folic acid AKI model, tubular cell culture with TWEAK treatment","journal":"Proceedings of the National Academy of Sciences","confidence":"High","confidence_rationale":"Tier 2 — multiple genetic knockouts and pharmacological inhibitors with distinct temporal dissection of cell death pathways in vivo and in vitro","pmids":["29588419"],"is_preprint":false},{"year":2018,"finding":"TRAF3IP2 mediates TWEAK/Fn14 pro-fibrotic signaling in cardiac fibroblasts: exogenous TWEAK upregulates TRAF3IP2 in a time- and dose-dependent manner (positive feedback), and TWEAK promotes TRAF3IP2 nuclear translocation. Silencing TRAF3IP2 inhibits TWEAK-induced p38 MAPK, NF-κB and AP-1 activation, inflammatory cytokine expression, MMP activation, collagen synthesis, and CF proliferation/migration. Genetic TRAF3IP2 ablation in vivo blocks TWEAK-induced cardiac fibrosis and dysfunction.","method":"TRAF3IP2 overexpression and siRNA knockdown in cardiac fibroblasts, p38 MAPK/NF-κB/AP-1 activation assays, collagen secretion assays, TWEAK infusion in mice, TRAF3IP2 knockout mice","journal":"Journal of molecular and cellular cardiology","confidence":"High","confidence_rationale":"Tier 2 — genetic KO in vivo and siRNA in vitro with multiple signaling pathway readouts demonstrating TRAF3IP2 as causal mediator","pmids":["29981796"],"is_preprint":false},{"year":2004,"finding":"A short variant of TWEAK (shortTWEAK) forms a complex with GSK-3β in a yeast two-hybrid system and colocalizes with GSK-3β in the nucleus of neuroblastoma cells. TWEAK is internalized and translocates to the nucleus, causing IκBα degradation, nuclear translocation of both GSK-3β and p65, and NF-κB-driven gene expression. LiCl (GSK-3β inhibitor) counteracts TWEAK-induced IL-8 expression.","method":"Yeast two-hybrid, colocalization by immunofluorescence, TWEAK internalization assay, NF-κB reporter assays, LiCl inhibition of GSK-3β","journal":"FEBS letters","confidence":"Medium","confidence_rationale":"Tier 2–3 — yeast two-hybrid with functional pharmacological validation, but nuclear TWEAK mechanism not extensively replicated","pmids":["15147869"],"is_preprint":false},{"year":2019,"finding":"TWEAK/Fn14 signaling drives VSMC proliferation and migration through upregulation of cyclins D1, CDK4/CDK6, and downregulation of CDK inhibitor p15INK4B via ERK and Akt activation. Genetic depletion of Tnfrsf12a or Tnfsf12 and pharmacological anti-TWEAK antibody reduce neointimal formation, cell proliferation, cyclin D1/CDK4/6, and increase p15INK4B in wire-injured mouse femoral arteries.","method":"Next-generation sequencing of TWEAK-regulated genes, Tnfrsf12a/Tnfsf12 knockout VSMCs, wound-healing/Transwell migration assays, cyclin/CDK expression by Western blot, wire injury mouse model, anti-TWEAK mAb treatment, ERK/Akt inhibition","journal":"EBioMedicine","confidence":"High","confidence_rationale":"Tier 2 — genetic KO in vitro and in vivo with mechanistic cell cycle pathway dissection and pharmacological blockade","pmids":["31395500"],"is_preprint":false},{"year":2012,"finding":"TWEAK/Fn14 activates NF-κB in cardiac fibroblasts, increasing MMP-9 production and promoting CF proliferation and collagen synthesis; siRNA knockdown of Fn14 or NF-κB inhibitor (PDTC) prevents these effects.","method":"Fn14 siRNA knockdown, PDTC NF-κB inhibition, MMP-9 activity assay, collagen synthesis measurement, CF proliferation assays","journal":"Molecular biology reports","confidence":"Medium","confidence_rationale":"Tier 2–3 — siRNA with multiple functional readouts but single lab study","pmids":["22555979"],"is_preprint":false},{"year":2021,"finding":"In keratinocytes, IFN-γ suppresses miR-149, and the TWEAK receptor Fn14 is a direct target of miR-149 (validated by 3'UTR reporter assays). IFN-γ-mediated miR-149 suppression amplifies keratinocyte inflammatory responses to TWEAK; synthetic miR-149 overexpression is protective in imiquimod-induced psoriasis mouse model.","method":"miR-149 target validation (reporter assays), miR-149 overexpression/inhibition in keratinocytes, intradermal miR-149 injection in mice, imiquimod psoriasis model","journal":"The Journal of allergy and clinical immunology","confidence":"Medium","confidence_rationale":"Tier 2 — direct target validation of Fn14 as miR-149 target with in vivo functional validation in psoriasis model","pmids":["33705829"],"is_preprint":false},{"year":2021,"finding":"In keratinocytes, TWEAK/Fn14 signaling synergizes with TNF and IL-17A to upregulate psoriasis-signature genes (CXC chemokines, IL-23A, IL-36G, S100A8/9, SERPINB1/B9). Keratinocyte-specific deletion of Fn14 reduces imiquimod-induced skin inflammation, epidermal hyperplasia, and psoriasis gene expression.","method":"Keratinocyte-specific Fn14 knockout mice, transcriptomic analysis in human keratinocytes, anti-TWEAK therapeutic treatment, imiquimod psoriasis model, combination cytokine stimulation assays","journal":"Science immunology","confidence":"High","confidence_rationale":"Tier 2 — cell-type-specific genetic KO with transcriptomic mechanistic data and in vivo phenotypic validation","pmids":["34797693"],"is_preprint":false},{"year":2018,"finding":"TWEAK/Fn14 signaling mediates HL-1 atrial myocyte hypertrophy through the JAK2/STAT3 pathway: TWEAK increases ANP and Troponin T expression, Fn14 knockdown blocks these effects, and siRNA inhibition of JAK2 or STAT3 attenuates TWEAK-induced hypertrophy.","method":"Fn14 knockdown, JAK2/STAT3 siRNA, hypertrophy markers (ANP, Troponin T) measurement in HL-1 atrial myocytes","journal":"Journal of cellular and molecular medicine","confidence":"Medium","confidence_rationale":"Tier 2–3 — siRNA knockdown with hypertrophy markers but single lab study in a cell line","pmids":["29971943"],"is_preprint":false},{"year":2016,"finding":"TWEAK upregulates ECE-1 (endothelin-converting enzyme-1) and ET-1 production in endothelial cells via AP-1 (through ERK1/2 and JNK) and NF-κB transcription factors acting on the ECE-1 promoter. Both AP-1 and NF-κB inhibitors partially block the effect. Validated in vivo: TWEAK injection in mice increases aortic and lung ECE-1 and ET-1 expression.","method":"ECE-1 promoter deletion/transfection assays, EMSA for AP-1 and NF-κB, ERK/JNK inhibitors, NF-κB inhibitor, in vivo TWEAK injection in mice, Western blot and qPCR, ET-1 ELISA","journal":"Cardiovascular research","confidence":"Medium","confidence_rationale":"Tier 2 — promoter dissection with EMSA and pharmacological inhibitors, confirmed in vivo","pmids":["28025386"],"is_preprint":false},{"year":2024,"finding":"TWEAK secreted by Th17 cells promotes epithelial-mesenchymal transition (EMT) in colorectal cancer cells by binding to Fn14 on tumor cells, enhancing tumor migration and invasion. In mouse models, Fn14 knockout (CRISPR) or Tnfsf12 deletion reduces liver metastases and prolongs survival. Cotransfer of Th17 cells with tumor cells promotes liver metastasis.","method":"Single-cell RNA sequencing, CRISPR-mediated Fn14 knockout, Tnfsf12 knockout mice, LNP-encapsulated siRNA, Il17a knockout mice, Th17 cotransfer experiments, EMT markers","journal":"Cancer research","confidence":"High","confidence_rationale":"Tier 2 — multiple genetic loss-of-function approaches with clear in vivo metastasis phenotype and mechanistic EMT readout","pmids":["38335276"],"is_preprint":false},{"year":2011,"finding":"IL-13-induced apoptosis (caspase-3 activation) in intestinal epithelial explants requires TWEAK, Fn14, and endogenous TNF-α secreted via ADAM17. Conversely, TNF-α-induced caspase activation requires IL-13, TWEAK, and Fn14. This was demonstrated using intestinal explants from TWEAK knockout mice.","method":"TWEAK knockout mice intestinal explants, ADAM17 inhibition, caspase-3 activation assays, IL-13 and TNF-α treatments","journal":"Gastroenterology","confidence":"High","confidence_rationale":"Tier 2 — genetic KO with mechanistic dissection of cytokine crosstalk and identified ADAM17-dependent pathway","pmids":["21893119"],"is_preprint":false},{"year":2021,"finding":"TWEAK administration induces cystogenesis and worsens the ADPKD phenotype; anti-TWEAK antibodies slow cystic progression and preserve renal function. The anti-cystogenic effect is associated with decreased MAPK signaling (cell proliferation-related), decreased NF-κB pathway activation, reduced fibrosis and apoptosis, and indirect decrease of macrophage recruitment.","method":"TWEAK administration in murine ADPKD model, anti-TWEAK antibody treatment, MAPK/NF-κB signaling assays, macrophage infiltration quantification, survival analysis","journal":"Journal of the American Society of Nephrology","confidence":"Medium","confidence_rationale":"Tier 2–3 — pharmacological gain- and loss-of-function in vivo with pathway readouts but no genetic KO confirmation","pmids":["34155062"],"is_preprint":false}],"current_model":"TNFSF12 (TWEAK) is a TNF superfamily cytokine that exists as membrane-anchored (processed intracellularly by furin at residues 90–93) and soluble isoforms; it binds its primary receptor Fn14 (TNFRSF12A) with ~2.3 nM affinity, recruiting TRAF1/2/3 to activate both canonical (rapid IκBα phosphorylation, RelA/p50) and noncanonical (IKK1/NIK-dependent p100 processing, RelB) NF-κB pathways; additionally, TWEAK transactivates EGFR via ADAM17-mediated shedding of HB-EGF/TGFα to drive ERK-dependent inflammation, upregulates ECE-1/ET-1 via AP-1 and NF-κB, and can signal through GSK-3β in the nucleus; soluble TWEAK is scavenged by CD163 acting as a decoy receptor; cell-type-specific responses include promotion of progenitor/myoblast proliferation via noncanonical NF-κB, induction of cathepsin B-dependent necrosis or caspase-dependent apoptosis, RIPK1/RIPK3/MLKL-dependent necroptosis in renal tubular cells, epigenetic downregulation of Klotho and PGC-1α via RelA-driven histone deacetylation, and promotion of EMT in tumor cells, collectively placing TWEAK/Fn14 as a multifunctional inflammatory axis regulating tissue repair, muscle biology, renal injury, and cancer progression."},"narrative":{"teleology":[{"year":2000,"claim":"Establishing that TWEAK is a monocyte-surface cytokine and ruling out DR3 as its receptor resolved a receptor-identity question that had complicated the field since TWEAK's cloning.","evidence":"Flow cytometry on IFN-γ-stimulated monocytes with anti-TWEAK mAb blocking; DR3-KO mouse cells retaining TWEAK binding","pmids":["11067885","11094155"],"confidence":"High","gaps":["Identity of the true TWEAK receptor was still unknown","Signaling pathways downstream of TWEAK binding uncharacterized"]},{"year":2001,"claim":"Identification of Fn14 as the bona fide TWEAK receptor with 2.3 nM affinity and TRAF1/2/3 recruitment established the ligand–receptor pair and immediately linked it to angiogenesis and endothelial growth.","evidence":"Receptor cloning from HUVEC library, saturation binding, Co-IP/pulldown for TRAF binding, corneal angiogenesis model","pmids":["11728344"],"confidence":"High","gaps":["Which NF-κB pathways are activated downstream of TRAF recruitment was unresolved","Relative roles of membrane vs. soluble TWEAK unknown"]},{"year":2002,"claim":"Dissection of TWEAK-induced cell death revealed that it operates through at least three distinct mechanisms — caspase-dependent apoptosis, cathepsin B-dependent necrosis, and indirect TNF-α-mediated killing — establishing TWEAK as a context-dependent death ligand.","evidence":"Caspase/cathepsin B inhibitors, lysosomal fractionation, anti-TNF-α blocking in multiple cell lines","pmids":["11777967"],"confidence":"High","gaps":["Receptor mediating TWEAK death signaling not yet confirmed as Fn14","Necroptotic pathway not yet explored"]},{"year":2003,"claim":"Demonstration that TWEAK/Fn14 sequentially activates both canonical (NEMO-dependent, rapid IκBα phosphorylation) and noncanonical (IKK1/NIK-dependent p100 processing) NF-κB defined the dual signaling architecture downstream of Fn14.","evidence":"TRAF-binding site mutagenesis, TRAF2/TRAF5-KO cells, proteasome inhibitor studies, EMSA","pmids":["12840022"],"confidence":"High","gaps":["Cell-type-specific dominance of canonical vs. noncanonical pathways undetermined","Non-NF-κB signaling arms not yet mapped"]},{"year":2005,"claim":"Showing that TWEAK/Fn14 selectively drives liver progenitor (oval) cell proliferation in vivo provided the first genetic evidence that TWEAK acts as a tissue-repair mitogen for progenitor populations.","evidence":"Fn14-KO mice, TWEAK-transgenic and adenoviral overexpression, anti-TWEAK mAb, DDC injury model","pmids":["16110324"],"confidence":"High","gaps":["Downstream signaling pathway mediating oval cell proliferation not fully resolved","Whether TWEAK drives other stem/progenitor compartments unknown"]},{"year":2006,"claim":"Extension to myoblasts and osteoblasts revealed that TWEAK/Fn14 broadly controls mesenchymal progenitor proliferation and differentiation, engaging PI3K–Akt and MAPK–ERK in addition to NF-κB depending on cell type.","evidence":"Fn14-KO primary myoblasts, C2C12 differentiation, cardiotoxin injury model, PI3K/ERK inhibitors in osteoblasts","pmids":["17124496","16945157"],"confidence":"High","gaps":["Mechanism by which TWEAK inhibits osteoblast differentiation vs. promoting myoblast proliferation not reconciled","Relative contribution of canonical vs. noncanonical NF-κB to myoblast phenotype unclear"]},{"year":2008,"claim":"In vivo renal studies established that TWEAK/Fn14 drives tubular NF-κB activation, chemokine production, and macrophage infiltration, positioning the axis as a central mediator of kidney inflammatory injury.","evidence":"Systemic TWEAK administration in mice, anti-TWEAK mAb in folic acid AKI, parthenolide NF-κB inhibition","pmids":["18235096"],"confidence":"High","gaps":["Direct vs. indirect effects on macrophage recruitment not separated","Whether non-NF-κB pathways contribute to renal injury not explored"]},{"year":2010,"claim":"Identification of furin cleavage at residues 90–93 and demonstration that uncleaved membrane TWEAK signals in trans via juxtacrine Fn14 engagement resolved how TWEAK exists in two functional forms.","evidence":"Furin cleavage-site mutagenesis, furin inhibitor, co-culture NF-κB reporter assays","pmids":["20385556"],"confidence":"High","gaps":["Whether other proteases contribute to shedding in specific tissues unknown","Relative potency of membrane vs. soluble TWEAK in vivo not quantified"]},{"year":2011,"claim":"Discovery that TWEAK epigenetically silences renal Klotho via RelA binding and histone deacetylation at its promoter revealed a chromatin-level mechanism by which TWEAK accelerates kidney aging/injury phenotypes.","evidence":"ChIP showing RelA at Klotho promoter, HDAC inhibitor rescue, TWEAK-KO mice, anti-TWEAK in folic acid AKI","pmids":["21719790"],"confidence":"High","gaps":["Which specific HDAC is recruited not identified","Whether Klotho downregulation is the primary driver of renal pathology vs. a parallel effect unclear"]},{"year":2012,"claim":"Selective genetic dissection showed that low-dose TWEAK promotes myoblast fusion specifically through the noncanonical NF-κB pathway (p100/RelB/IKKα/NIK), while high-dose TWEAK engages canonical NF-κB causing atrophy, resolving a dose-dependent signaling paradox.","evidence":"cIAP1-KO, siRNA of p100/RelB/IKKα/NIK/TRAF3, p52/RelB overexpression, myotube fusion assays","pmids":["23074266"],"confidence":"High","gaps":["In vivo confirmation of dose-dependent pathway switching not performed","Whether cIAP1 degradation is a physiological TWEAK-triggered event unclear"]},{"year":2013,"claim":"Identification of ADAM17-mediated EGFR transactivation as an EGFR-dependent, NF-κB-independent arm of TWEAK signaling revealed a second major effector pathway that drives ERK-dependent inflammation through HB-EGF and TGFα shedding.","evidence":"ADAM17 inhibitor, erlotinib, EGFR phosphorylation assays, in vivo TWEAK administration","pmids":["24037740"],"confidence":"High","gaps":["Whether EGFR transactivation occurs in non-renal tissues unknown","Direct mechanism linking Fn14 to ADAM17 activation not defined"]},{"year":2015,"claim":"Establishing CD163 as a decoy/scavenger receptor for soluble TWEAK explained how macrophage phenotype regulates TWEAK bioavailability and muscle progenitor cell responses during ischemia.","evidence":"CD163-KO mice with elevated TWEAK and enhanced satellite cell proliferation, binding assays, ischemic injury model","pmids":["26242746","19473660"],"confidence":"High","gaps":["Whether CD163 signals upon TWEAK binding or is purely a decoy not resolved","Contribution of CD163-TWEAK axis in non-muscle tissues not explored"]},{"year":2016,"claim":"Extension of the epigenetic silencing mechanism to PGC-1α — via NF-κB-driven histone deacetylation at its promoter — linked TWEAK to mitochondrial dysfunction in both muscle and kidney, unifying metabolic and inflammatory consequences of sustained TWEAK signaling.","evidence":"ChIP at PGC-1α promoter, HDAC/NF-κB inhibitors, adenoviral PGC-1α rescue, TWEAK-KO/Fn14-KO mice denervation model","pmids":["26535995","24327607"],"confidence":"High","gaps":["Identity of the recruited HDAC enzyme not determined","Whether TWEAK affects other metabolic gene promoters via the same mechanism unknown"]},{"year":2018,"claim":"Demonstration that TWEAK/Fn14 triggers RIPK1/RIPK3/MLKL-dependent necroptosis as a temporally distinct late wave of tubular cell death in AKI established necroptosis as a TWEAK-mediated death modality and explained why caspase inhibition alone is insufficient for renal protection.","evidence":"Necrostatin-1, RIPK3-deficient and MLKL-deficient mice, Fn14-KO, folic acid AKI with temporal dissection","pmids":["29588419"],"confidence":"High","gaps":["Whether TWEAK directly activates RIPK1 or does so via TNF-α co-signaling not fully resolved","Upstream signals determining apoptosis vs. necroptosis switching remain unclear"]},{"year":2018,"claim":"Identification of TRAF3IP2 as a required adaptor for TWEAK-induced cardiac fibrosis — mediating p38 MAPK, NF-κB, and AP-1 activation — revealed a tissue-specific signaling node in fibroblasts.","evidence":"TRAF3IP2-KO mice, siRNA in cardiac fibroblasts, TWEAK infusion, collagen/MMP readouts","pmids":["29981796"],"confidence":"High","gaps":["How TRAF3IP2 is recruited to the Fn14 signaling complex not determined","Whether TRAF3IP2 participates in TWEAK signaling in non-cardiac fibroblasts unknown"]},{"year":2019,"claim":"Mechanistic dissection of VSMC proliferation showed TWEAK drives cell cycle progression via ERK/Akt-dependent cyclin D1–CDK4/6 upregulation and p15INK4B suppression, explaining TWEAK's pro-neointimal role after vascular injury.","evidence":"Tnfrsf12a/Tnfsf12-KO VSMCs, wire injury model, anti-TWEAK mAb, ERK/Akt inhibitors, NGS","pmids":["31395500"],"confidence":"High","gaps":["Whether noncanonical NF-κB contributes to VSMC phenotype not tested","Relevance to human restenosis not established"]},{"year":2021,"claim":"TWEAK/Fn14 was shown to synergize with TNF and IL-17A in keratinocytes to amplify psoriasis-signature gene expression, and keratinocyte-specific Fn14 deletion protected against psoriasiform inflammation, establishing TWEAK as a co-driver of skin autoimmunity.","evidence":"Keratinocyte-specific Fn14-KO mice, imiquimod model, transcriptomics, miR-149 target validation for Fn14","pmids":["34797693","33705829"],"confidence":"High","gaps":["Whether TWEAK blockade is therapeutically effective in human psoriasis untested","Mechanism of TWEAK synergy with IL-17A at the promoter level not defined"]},{"year":2024,"claim":"Th17-derived TWEAK was identified as a driver of colorectal cancer liver metastasis through Fn14-dependent EMT, demonstrating a tumor-extrinsic immune source of TWEAK that promotes cancer dissemination.","evidence":"scRNA-seq, CRISPR Fn14-KO tumor cells, Tnfsf12-KO mice, Th17 cotransfer experiments, EMT marker analysis","pmids":["38335276"],"confidence":"High","gaps":["Whether TWEAK-induced EMT is reversible upon Fn14 blockade not addressed","Applicability beyond colorectal cancer not explored","Contribution of other Th17 cytokines confounding in cotransfer experiments"]},{"year":null,"claim":"Major unresolved questions include the structural basis of Fn14 signaling complex assembly, the identity of HDACs recruited to TWEAK-silenced promoters, the molecular switch determining whether TWEAK triggers apoptosis, necroptosis, or proliferation in a given cell, and the therapeutic window for TWEAK/Fn14 blockade in human disease.","evidence":"","pmids":[],"confidence":"Low","gaps":["No crystal structure of TWEAK–Fn14 signaling complex available","HDAC identity at Klotho and PGC-1α promoters not determined","Dose- and context-dependent pathway switching mechanism not molecularly defined"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0048018","term_label":"receptor ligand activity","supporting_discovery_ids":[0,3,6,7,8,27]},{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[1,12,13]}],"localization":[{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[3,9]},{"term_id":"GO:0005576","term_label":"extracellular region","supporting_discovery_ids":[9,14]}],"pathway":[{"term_id":"R-HSA-168256","term_label":"Immune System","supporting_discovery_ids":[3,8,24]},{"term_id":"R-HSA-5357801","term_label":"Programmed Cell Death","supporting_discovery_ids":[2,18,28]},{"term_id":"R-HSA-1643685","term_label":"Disease","supporting_discovery_ids":[27,29]}],"complexes":[],"partners":["TNFRSF12A","TRAF1","TRAF2","TRAF3","CD163","TRAF3IP2","ADAM17","GSK3B"],"other_free_text":[]},"mechanistic_narrative":"TNFSF12 (TWEAK) is a TNF superfamily cytokine that functions as a pleiotropic regulator of inflammation, tissue repair, cell death, and proliferation through engagement of its receptor Fn14 (TNFRSF12A). TWEAK exists as a membrane-anchored form processed intracellularly by furin at residues 90–93 and as a soluble form scavenged by the decoy receptor CD163; membrane TWEAK signals in a juxtacrine manner, while soluble TWEAK acts at a distance, with both forms activating canonical (IκBα/RelA/p50) and noncanonical (NIK/IKK1/p100→p52/RelB) NF-κB pathways through TRAF1/2/3 recruitment to the Fn14 cytoplasmic domain [PMID:11728344, PMID:12840022, PMID:20385556, PMID:26242746]. Beyond NF-κB, TWEAK/Fn14 transactivates EGFR via ADAM17-mediated shedding of HB-EGF and TGFα to drive ERK-dependent inflammatory gene expression, activates PI3K–Akt and MAPK–ERK cascades regulating cell cycle progression, and epigenetically silences protective genes including Klotho and PGC-1α through RelA-directed histone deacetylation at their promoters [PMID:24037740, PMID:21719790, PMID:26535995, PMID:31395500]. These signaling outputs underlie context-dependent biological activities: stimulation of progenitor and myoblast proliferation and fusion via noncanonical NF-κB, induction of cathepsin B–dependent necrosis or caspase-dependent apoptosis, RIPK1/RIPK3/MLKL-dependent necroptosis in renal tubular cells during acute kidney injury, promotion of epithelial–mesenchymal transition driving colorectal cancer metastasis, and pro-fibrotic remodeling in cardiac tissue through the adaptor TRAF3IP2 [PMID:23074266, PMID:11777967, PMID:29588419, PMID:38335276, PMID:29981796]."},"prefetch_data":{"uniprot":{"accession":"O43508","full_name":"Tumor necrosis factor ligand superfamily member 12","aliases":["APO3 ligand","TNF-related weak inducer of apoptosis","TWEAK"],"length_aa":249,"mass_kda":27.2,"function":"Binds to FN14 and possibly also to TNRFSF12/APO3. Weak inducer of apoptosis in some cell types. Mediates NF-kappa-B activation. Promotes angiogenesis and the proliferation of endothelial cells. Also involved in induction of inflammatory cytokines. Promotes IL8 secretion","subcellular_location":"Cell membrane","url":"https://www.uniprot.org/uniprotkb/O43508/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/TNFSF12","classification":"Not Classified","n_dependent_lines":0,"n_total_lines":1208,"dependency_fraction":0.0},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/TNFSF12","total_profiled":1310},"omim":[{"mim_id":"608464","title":"ANGIOGENIC FACTOR WITH G-PATCH AND FHA DOMAINS 1; AGGF1","url":"https://www.omim.org/entry/608464"},{"mim_id":"605914","title":"TUMOR NECROSIS FACTOR RECEPTOR SUPERFAMILY, MEMBER 12A; TNFRSF12A","url":"https://www.omim.org/entry/605914"},{"mim_id":"604472","title":"TUMOR NECROSIS FACTOR LIGAND SUPERFAMILY, MEMBER 13; TNFSF13","url":"https://www.omim.org/entry/604472"},{"mim_id":"603366","title":"TUMOR NECROSIS FACTOR RECEPTOR SUPERFAMILY, MEMBER 25; TNFRSF25","url":"https://www.omim.org/entry/603366"},{"mim_id":"602695","title":"TUMOR NECROSIS FACTOR LIGAND SUPERFAMILY, MEMBER 12; TNFSF12","url":"https://www.omim.org/entry/602695"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Approved","locations":[{"location":"Vesicles","reliability":"Approved"}],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in many","driving_tissues":[],"url":"https://www.proteinatlas.org/search/TNFSF12"},"hgnc":{"alias_symbol":["TWEAK","DR3LG","APO3L","TNF12"],"prev_symbol":[]},"alphafold":{"accession":"O43508","domains":[{"cath_id":"2.60.120.40","chopping":"108-246","consensus_level":"high","plddt":93.2414,"start":108,"end":246}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/O43508","model_url":"https://alphafold.ebi.ac.uk/files/AF-O43508-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-O43508-F1-predicted_aligned_error_v6.png","plddt_mean":77.88},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=TNFSF12","jax_strain_url":"https://www.jax.org/strain/search?query=TNFSF12"},"sequence":{"accession":"O43508","fasta_url":"https://rest.uniprot.org/uniprotkb/O43508.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/O43508/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/O43508"}},"corpus_meta":[{"pmid":"18404150","id":"PMC_18404150","title":"The TWEAK-Fn14 cytokine-receptor axis: discovery, biology and therapeutic targeting.","date":"2008","source":"Nature reviews. Drug discovery","url":"https://pubmed.ncbi.nlm.nih.gov/18404150","citation_count":492,"is_preprint":false},{"pmid":"21719790","id":"PMC_21719790","title":"The inflammatory cytokines TWEAK and TNFα reduce renal klotho expression through NFκB.","date":"2011","source":"Journal of the American Society of Nephrology : JASN","url":"https://pubmed.ncbi.nlm.nih.gov/21719790","citation_count":343,"is_preprint":false},{"pmid":"16110324","id":"PMC_16110324","title":"TWEAK induces liver progenitor cell proliferation.","date":"2005","source":"The Journal of clinical investigation","url":"https://pubmed.ncbi.nlm.nih.gov/16110324","citation_count":334,"is_preprint":false},{"pmid":"11728344","id":"PMC_11728344","title":"A novel TNF receptor family member binds TWEAK and is implicated in angiogenesis.","date":"2001","source":"Immunity","url":"https://pubmed.ncbi.nlm.nih.gov/11728344","citation_count":327,"is_preprint":false},{"pmid":"12840022","id":"PMC_12840022","title":"TWEAK induces NF-kappaB2 p100 processing and long lasting NF-kappaB activation.","date":"2003","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/12840022","citation_count":277,"is_preprint":false},{"pmid":"12787562","id":"PMC_12787562","title":"TWEAK, a member of the TNF superfamily, is a multifunctional cytokine that binds the TweakR/Fn14 receptor.","date":"2003","source":"Cytokine & growth factor reviews","url":"https://pubmed.ncbi.nlm.nih.gov/12787562","citation_count":253,"is_preprint":false},{"pmid":"17981048","id":"PMC_17981048","title":"TWEAKing tissue remodeling by a multifunctional cytokine: role of TWEAK/Fn14 pathway in health and disease.","date":"2007","source":"Cytokine","url":"https://pubmed.ncbi.nlm.nih.gov/17981048","citation_count":244,"is_preprint":false},{"pmid":"17124496","id":"PMC_17124496","title":"TWEAK, via its receptor Fn14, is a novel regulator of mesenchymal progenitor cells and skeletal muscle regeneration.","date":"2006","source":"The EMBO journal","url":"https://pubmed.ncbi.nlm.nih.gov/17124496","citation_count":192,"is_preprint":false},{"pmid":"18235096","id":"PMC_18235096","title":"The cytokine TWEAK modulates renal tubulointerstitial inflammation.","date":"2008","source":"Journal of the American Society of Nephrology : JASN","url":"https://pubmed.ncbi.nlm.nih.gov/18235096","citation_count":164,"is_preprint":false},{"pmid":"11777967","id":"PMC_11777967","title":"Multiple pathways of TWEAK-induced cell death.","date":"2002","source":"Journal of immunology (Baltimore, Md. : 1950)","url":"https://pubmed.ncbi.nlm.nih.gov/11777967","citation_count":148,"is_preprint":false},{"pmid":"11067885","id":"PMC_11067885","title":"Involvement of TWEAK in interferon gamma-stimulated monocyte cytotoxicity.","date":"2000","source":"The Journal of experimental medicine","url":"https://pubmed.ncbi.nlm.nih.gov/11067885","citation_count":146,"is_preprint":false},{"pmid":"16424220","id":"PMC_16424220","title":"Proinflammatory effects of TWEAK/Fn14 interactions in glomerular mesangial cells.","date":"2006","source":"Journal of immunology (Baltimore, Md. : 1950)","url":"https://pubmed.ncbi.nlm.nih.gov/16424220","citation_count":139,"is_preprint":false},{"pmid":"29588419","id":"PMC_29588419","title":"TWEAK and RIPK1 mediate a second wave of cell death during AKI.","date":"2018","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/29588419","citation_count":129,"is_preprint":false},{"pmid":"19473660","id":"PMC_19473660","title":"The CD163-expressing macrophages recognize and internalize TWEAK: potential consequences in atherosclerosis.","date":"2009","source":"Atherosclerosis","url":"https://pubmed.ncbi.nlm.nih.gov/19473660","citation_count":124,"is_preprint":false},{"pmid":"15056843","id":"PMC_15056843","title":"TWEAK can induce pro-inflammatory cytokines and matrix metalloproteinase-9 in macrophages.","date":"2004","source":"Circulation journal : official journal of the Japanese Circulation Society","url":"https://pubmed.ncbi.nlm.nih.gov/15056843","citation_count":123,"is_preprint":false},{"pmid":"12411489","id":"PMC_12411489","title":"An endogenous hybrid mRNA encodes TWE-PRIL, a functional cell surface TWEAK-APRIL fusion protein.","date":"2002","source":"The EMBO journal","url":"https://pubmed.ncbi.nlm.nih.gov/12411489","citation_count":117,"is_preprint":false},{"pmid":"26535995","id":"PMC_26535995","title":"The inflammatory cytokine TWEAK decreases PGC-1α expression and mitochondrial function in acute kidney injury.","date":"2016","source":"Kidney international","url":"https://pubmed.ncbi.nlm.nih.gov/26535995","citation_count":116,"is_preprint":false},{"pmid":"11839778","id":"PMC_11839778","title":"Dual role for TWEAK in angiogenic regulation.","date":"2002","source":"Journal of cell science","url":"https://pubmed.ncbi.nlm.nih.gov/11839778","citation_count":108,"is_preprint":false},{"pmid":"10706675","id":"PMC_10706675","title":"TRAIL (Apo2 ligand) and TWEAK (Apo3 ligand) mediate CD4+ T cell killing of antigen-presenting macrophages.","date":"2000","source":"Journal of immunology (Baltimore, Md. : 1950)","url":"https://pubmed.ncbi.nlm.nih.gov/10706675","citation_count":108,"is_preprint":false},{"pmid":"18945991","id":"PMC_18945991","title":"Additive effects of soluble TWEAK and inflammation on mortality in hemodialysis patients.","date":"2008","source":"Clinical journal of the American Society of Nephrology : CJASN","url":"https://pubmed.ncbi.nlm.nih.gov/18945991","citation_count":100,"is_preprint":false},{"pmid":"15353286","id":"PMC_15353286","title":"The role of TWEAK/Fn14 in the pathogenesis of inflammation and systemic autoimmunity.","date":"2004","source":"Frontiers in bioscience : a journal and virtual library","url":"https://pubmed.ncbi.nlm.nih.gov/15353286","citation_count":97,"is_preprint":false},{"pmid":"22727560","id":"PMC_22727560","title":"Role of TWEAK in lupus nephritis: a bench-to-bedside review.","date":"2012","source":"Journal of autoimmunity","url":"https://pubmed.ncbi.nlm.nih.gov/22727560","citation_count":95,"is_preprint":false},{"pmid":"26242746","id":"PMC_26242746","title":"CD163 interacts with TWEAK to regulate tissue regeneration after ischaemic injury.","date":"2015","source":"Nature communications","url":"https://pubmed.ncbi.nlm.nih.gov/26242746","citation_count":91,"is_preprint":false},{"pmid":"28619231","id":"PMC_28619231","title":"Endoplasmic reticulum chaperones tweak the mitochondrial calcium rheostat to control metabolism and cell death.","date":"2017","source":"Cell calcium","url":"https://pubmed.ncbi.nlm.nih.gov/28619231","citation_count":86,"is_preprint":false},{"pmid":"22982296","id":"PMC_22982296","title":"Inhibition of the TWEAK/Fn14 pathway attenuates renal disease in nephrotoxic serum nephritis.","date":"2012","source":"Clinical immunology (Orlando, Fla.)","url":"https://pubmed.ncbi.nlm.nih.gov/22982296","citation_count":86,"is_preprint":false},{"pmid":"21330486","id":"PMC_21330486","title":"Soluble TWEAK and PTX3 in nondialysis CKD patients: impact on endothelial dysfunction and cardiovascular outcomes.","date":"2011","source":"Clinical journal of the American Society of Nephrology : CJASN","url":"https://pubmed.ncbi.nlm.nih.gov/21330486","citation_count":85,"is_preprint":false},{"pmid":"28809783","id":"PMC_28809783","title":"Role of Omentin, Vaspin, Cardiotrophin-1, TWEAK and NOV/CCN3 in Obesity and Diabetes Development.","date":"2017","source":"International journal of molecular sciences","url":"https://pubmed.ncbi.nlm.nih.gov/28809783","citation_count":83,"is_preprint":false},{"pmid":"24400009","id":"PMC_24400009","title":"Neuropsychiatric Lupus, the Blood Brain Barrier, and the TWEAK/Fn14 Pathway.","date":"2013","source":"Frontiers in immunology","url":"https://pubmed.ncbi.nlm.nih.gov/24400009","citation_count":80,"is_preprint":false},{"pmid":"34797693","id":"PMC_34797693","title":"TWEAK functions with TNF and IL-17 on keratinocytes and is a potential target for psoriasis therapy.","date":"2021","source":"Science immunology","url":"https://pubmed.ncbi.nlm.nih.gov/34797693","citation_count":77,"is_preprint":false},{"pmid":"21893119","id":"PMC_21893119","title":"Interleukin-13 damages intestinal mucosa via TWEAK and Fn14 in mice-a pathway associated with ulcerative colitis.","date":"2011","source":"Gastroenterology","url":"https://pubmed.ncbi.nlm.nih.gov/21893119","citation_count":74,"is_preprint":false},{"pmid":"23074266","id":"PMC_23074266","title":"TWEAK and cIAP1 regulate myoblast fusion through the noncanonical NF-κB signaling pathway.","date":"2012","source":"Science signaling","url":"https://pubmed.ncbi.nlm.nih.gov/23074266","citation_count":70,"is_preprint":false},{"pmid":"28530223","id":"PMC_28530223","title":"TWEAK mediates inflammation in experimental atopic dermatitis and psoriasis.","date":"2017","source":"Nature communications","url":"https://pubmed.ncbi.nlm.nih.gov/28530223","citation_count":68,"is_preprint":false},{"pmid":"20068083","id":"PMC_20068083","title":"Antibodies to TWEAK receptor inhibit human tumor growth through dual mechanisms.","date":"2010","source":"Clinical cancer research : an official journal of the American Association for Cancer Research","url":"https://pubmed.ncbi.nlm.nih.gov/20068083","citation_count":65,"is_preprint":false},{"pmid":"20385556","id":"PMC_20385556","title":"Full-length, membrane-anchored TWEAK can function as a juxtacrine signaling molecule and activate the NF-kappaB pathway.","date":"2010","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/20385556","citation_count":65,"is_preprint":false},{"pmid":"23320797","id":"PMC_23320797","title":"TWEAK/Fn14 pathway modulates properties of a human microvascular endothelial cell model of blood brain barrier.","date":"2013","source":"Journal of neuroinflammation","url":"https://pubmed.ncbi.nlm.nih.gov/23320797","citation_count":64,"is_preprint":false},{"pmid":"26300004","id":"PMC_26300004","title":"The TWEAK receptor Fn14 is a potential cell surface portal for targeted delivery of glioblastoma therapeutics.","date":"2015","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/26300004","citation_count":62,"is_preprint":false},{"pmid":"16945157","id":"PMC_16945157","title":"TWEAK/Fn14 interaction regulates RANTES production, BMP-2-induced differentiation, and RANKL expression in mouse osteoblastic MC3T3-E1 cells.","date":"2006","source":"Arthritis research & therapy","url":"https://pubmed.ncbi.nlm.nih.gov/16945157","citation_count":61,"is_preprint":false},{"pmid":"24478779","id":"PMC_24478779","title":"TWEAK/Fn14 Signaling Axis Mediates Skeletal Muscle Atrophy and Metabolic Dysfunction.","date":"2014","source":"Frontiers in immunology","url":"https://pubmed.ncbi.nlm.nih.gov/24478779","citation_count":60,"is_preprint":false},{"pmid":"24327607","id":"PMC_24327607","title":"Regulatory circuitry of TWEAK-Fn14 system and PGC-1α in skeletal muscle atrophy program.","date":"2013","source":"FASEB journal : official publication of the Federation of American Societies for Experimental Biology","url":"https://pubmed.ncbi.nlm.nih.gov/24327607","citation_count":59,"is_preprint":false},{"pmid":"38335276","id":"PMC_38335276","title":"Th17 Cells Secrete TWEAK to Trigger Epithelial-Mesenchymal Transition and Promote Colorectal Cancer Liver Metastasis.","date":"2024","source":"Cancer research","url":"https://pubmed.ncbi.nlm.nih.gov/38335276","citation_count":58,"is_preprint":false},{"pmid":"24444596","id":"PMC_24444596","title":"The TWEAK-Fn14 pathway: a potent regulator of skeletal muscle biology in health and disease.","date":"2013","source":"Cytokine & growth factor reviews","url":"https://pubmed.ncbi.nlm.nih.gov/24444596","citation_count":58,"is_preprint":false},{"pmid":"18483204","id":"PMC_18483204","title":"No end in site: TWEAK/Fn14 activation and autoimmunity associated- end-organ pathologies.","date":"2008","source":"Journal of leukocyte biology","url":"https://pubmed.ncbi.nlm.nih.gov/18483204","citation_count":58,"is_preprint":false},{"pmid":"22672874","id":"PMC_22672874","title":"The TWEAK/Fn14 pathway as an aggravating and perpetuating factor in inflammatory diseases: focus on inflammatory bowel diseases.","date":"2012","source":"Journal of leukocyte biology","url":"https://pubmed.ncbi.nlm.nih.gov/22672874","citation_count":56,"is_preprint":false},{"pmid":"33221352","id":"PMC_33221352","title":"TWEAK/Fn14 signalling promotes cholangiocarcinoma niche formation and progression.","date":"2020","source":"Journal of hepatology","url":"https://pubmed.ncbi.nlm.nih.gov/33221352","citation_count":55,"is_preprint":false},{"pmid":"28639899","id":"PMC_28639899","title":"TWEAK/Fn14 signaling in tumors.","date":"2017","source":"Tumour biology : the journal of the International Society for Oncodevelopmental Biology and Medicine","url":"https://pubmed.ncbi.nlm.nih.gov/28639899","citation_count":55,"is_preprint":false},{"pmid":"34542797","id":"PMC_34542797","title":"The TWEAK/Fn14/CD163 axis-implications for metabolic disease.","date":"2021","source":"Reviews in endocrine & metabolic disorders","url":"https://pubmed.ncbi.nlm.nih.gov/34542797","citation_count":55,"is_preprint":false},{"pmid":"19482507","id":"PMC_19482507","title":"Considering TWEAK as a target for therapy in renal and vascular injury.","date":"2009","source":"Cytokine & growth factor reviews","url":"https://pubmed.ncbi.nlm.nih.gov/19482507","citation_count":53,"is_preprint":false},{"pmid":"11112433","id":"PMC_11112433","title":"Down-regulated expression of TWEAK mRNA in acute and chronic inflammatory pathologies.","date":"2000","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/11112433","citation_count":52,"is_preprint":false},{"pmid":"22555979","id":"PMC_22555979","title":"TWEAK/Fn14 promotes the proliferation and collagen synthesis of rat cardiac fibroblasts via the NF-кB pathway.","date":"2012","source":"Molecular biology reports","url":"https://pubmed.ncbi.nlm.nih.gov/22555979","citation_count":52,"is_preprint":false},{"pmid":"12821115","id":"PMC_12821115","title":"Characterization of murine TWEAK and its receptor (Fn14) by monoclonal antibodies.","date":"2003","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/12821115","citation_count":51,"is_preprint":false},{"pmid":"30010468","id":"PMC_30010468","title":"The lncRNA RHPN1-AS1 downregulation promotes gefitinib resistance by targeting miR-299-3p/TNFSF12 pathway in NSCLC.","date":"2018","source":"Cell cycle (Georgetown, Tex.)","url":"https://pubmed.ncbi.nlm.nih.gov/30010468","citation_count":50,"is_preprint":false},{"pmid":"18793781","id":"PMC_18793781","title":"Induction of the cytokine TWEAK and its receptor Fn14 in ischemic stroke.","date":"2008","source":"Journal of the neurological sciences","url":"https://pubmed.ncbi.nlm.nih.gov/18793781","citation_count":49,"is_preprint":false},{"pmid":"33705829","id":"PMC_33705829","title":"Cross-talk between IFN-γ and TWEAK through miR-149 amplifies skin inflammation in psoriasis.","date":"2021","source":"The Journal of allergy and clinical immunology","url":"https://pubmed.ncbi.nlm.nih.gov/33705829","citation_count":49,"is_preprint":false},{"pmid":"17127278","id":"PMC_17127278","title":"Role of TWEAK and Fn14 in tumor biology.","date":"2007","source":"Frontiers in bioscience : a journal and virtual library","url":"https://pubmed.ncbi.nlm.nih.gov/17127278","citation_count":48,"is_preprint":false},{"pmid":"24970477","id":"PMC_24970477","title":"AR-regulated TWEAK-FN14 pathway promotes prostate cancer bone metastasis.","date":"2014","source":"Cancer research","url":"https://pubmed.ncbi.nlm.nih.gov/24970477","citation_count":48,"is_preprint":false},{"pmid":"23190886","id":"PMC_23190886","title":"The TWEAK receptor Fn14 is a therapeutic target in melanoma: immunotoxins targeting Fn14 receptor for malignant melanoma treatment.","date":"2012","source":"The Journal of investigative dermatology","url":"https://pubmed.ncbi.nlm.nih.gov/23190886","citation_count":47,"is_preprint":false},{"pmid":"11094155","id":"PMC_11094155","title":"Studies on the interaction between TWEAK and the death receptor WSL-1/TRAMP (DR3).","date":"2000","source":"FEBS letters","url":"https://pubmed.ncbi.nlm.nih.gov/11094155","citation_count":46,"is_preprint":false},{"pmid":"24037740","id":"PMC_24037740","title":"TWEAK transactivation of the epidermal growth factor receptor mediates renal inflammation.","date":"2013","source":"The Journal of pathology","url":"https://pubmed.ncbi.nlm.nih.gov/24037740","citation_count":46,"is_preprint":false},{"pmid":"24550918","id":"PMC_24550918","title":"Role of the TWEAK-Fn14-cIAP1-NF-κB Signaling Axis in the Regulation of Myogenesis and Muscle Homeostasis.","date":"2014","source":"Frontiers in immunology","url":"https://pubmed.ncbi.nlm.nih.gov/24550918","citation_count":43,"is_preprint":false},{"pmid":"18310134","id":"PMC_18310134","title":"Involvement of TWEAK/Fn14 interaction in the synovial inflammation of RA.","date":"2008","source":"Rheumatology (Oxford, England)","url":"https://pubmed.ncbi.nlm.nih.gov/18310134","citation_count":43,"is_preprint":false},{"pmid":"20189297","id":"PMC_20189297","title":"TWEAK/Fn14 promotes apoptosis of human endometrial cancer cells via caspase pathway.","date":"2010","source":"Cancer letters","url":"https://pubmed.ncbi.nlm.nih.gov/20189297","citation_count":41,"is_preprint":false},{"pmid":"24416031","id":"PMC_24416031","title":"TWEAK: A New Player in Obesity and Diabetes.","date":"2013","source":"Frontiers in immunology","url":"https://pubmed.ncbi.nlm.nih.gov/24416031","citation_count":39,"is_preprint":false},{"pmid":"15586228","id":"PMC_15586228","title":"Functional expression of TWEAK in human colonic adenocarcinoma cells.","date":"2005","source":"International journal of oncology","url":"https://pubmed.ncbi.nlm.nih.gov/15586228","citation_count":38,"is_preprint":false},{"pmid":"18945822","id":"PMC_18945822","title":"TWEAK is expressed at the cell surface of monocytes during multiple sclerosis.","date":"2008","source":"Journal of leukocyte biology","url":"https://pubmed.ncbi.nlm.nih.gov/18945822","citation_count":38,"is_preprint":false},{"pmid":"25539934","id":"PMC_25539934","title":"TWEAK-Fn14 pathway activation after exercise in human skeletal muscle: insights from two exercise modes and a time course investigation.","date":"2014","source":"Journal of applied physiology (Bethesda, Md. : 1985)","url":"https://pubmed.ncbi.nlm.nih.gov/25539934","citation_count":37,"is_preprint":false},{"pmid":"24479820","id":"PMC_24479820","title":"Genetic deletion or TWEAK blocking antibody administration reduce atherosclerosis and enhance plaque stability in mice.","date":"2014","source":"Journal of cellular and molecular medicine","url":"https://pubmed.ncbi.nlm.nih.gov/24479820","citation_count":36,"is_preprint":false},{"pmid":"24469836","id":"PMC_24469836","title":"Mcl-1 mediates TWEAK/Fn14-induced non-small cell lung cancer survival and therapeutic response.","date":"2014","source":"Molecular cancer research : MCR","url":"https://pubmed.ncbi.nlm.nih.gov/24469836","citation_count":36,"is_preprint":false},{"pmid":"15147869","id":"PMC_15147869","title":"Involvement of GSK-3beta in TWEAK-mediated NF-kappaB activation.","date":"2004","source":"FEBS letters","url":"https://pubmed.ncbi.nlm.nih.gov/15147869","citation_count":36,"is_preprint":false},{"pmid":"29456665","id":"PMC_29456665","title":"Involvement of TWEAK and the NF-κB signaling pathway in lupus nephritis.","date":"2018","source":"Experimental and therapeutic medicine","url":"https://pubmed.ncbi.nlm.nih.gov/29456665","citation_count":34,"is_preprint":false},{"pmid":"29981796","id":"PMC_29981796","title":"TRAF3IP2 mediates TWEAK/TWEAKR-induced pro-fibrotic responses in cultured cardiac fibroblasts and the heart.","date":"2018","source":"Journal of molecular and cellular cardiology","url":"https://pubmed.ncbi.nlm.nih.gov/29981796","citation_count":34,"is_preprint":false},{"pmid":"34155062","id":"PMC_34155062","title":"TWEAK Signaling Pathway Blockade Slows Cyst Growth and Disease Progression in Autosomal Dominant Polycystic Kidney Disease.","date":"2021","source":"Journal of the American Society of Nephrology : JASN","url":"https://pubmed.ncbi.nlm.nih.gov/34155062","citation_count":33,"is_preprint":false},{"pmid":"24611063","id":"PMC_24611063","title":"TWEAK-Fn14 Cytokine-Receptor Axis: A New Player of Myocardial Remodeling and Cardiac Failure.","date":"2014","source":"Frontiers in immunology","url":"https://pubmed.ncbi.nlm.nih.gov/24611063","citation_count":33,"is_preprint":false},{"pmid":"26659091","id":"PMC_26659091","title":"Role of the TWEAK/Fn14 pathway in autoimmune diseases.","date":"2016","source":"Immunologic research","url":"https://pubmed.ncbi.nlm.nih.gov/26659091","citation_count":32,"is_preprint":false},{"pmid":"25504044","id":"PMC_25504044","title":"TWEAK/Fn14, a pathway and novel therapeutic target in myotonic dystrophy.","date":"2014","source":"Human molecular genetics","url":"https://pubmed.ncbi.nlm.nih.gov/25504044","citation_count":32,"is_preprint":false},{"pmid":"27725864","id":"PMC_27725864","title":"TWEAK protects cardiomyocyte against apoptosis in a PI3K/AKT pathway dependent manner.","date":"2016","source":"American journal of translational research","url":"https://pubmed.ncbi.nlm.nih.gov/27725864","citation_count":31,"is_preprint":false},{"pmid":"27339384","id":"PMC_27339384","title":"Out of the TWEAKlight: Elucidating the Role of Fn14 and TWEAK in Acute Kidney Injury.","date":"2016","source":"Seminars in nephrology","url":"https://pubmed.ncbi.nlm.nih.gov/27339384","citation_count":30,"is_preprint":false},{"pmid":"32526219","id":"PMC_32526219","title":"TWEAK/Fn14 axis in respiratory diseases.","date":"2020","source":"Clinica chimica acta; international journal of clinical chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/32526219","citation_count":30,"is_preprint":false},{"pmid":"24376672","id":"PMC_24376672","title":"BAFF, APRIL, TWEAK, BCMA, TACI and Fn14 proteins are related to human glioma tumor grade: immunohistochemistry and public microarray data meta-analysis.","date":"2013","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/24376672","citation_count":30,"is_preprint":false},{"pmid":"31395500","id":"PMC_31395500","title":"A major role of TWEAK/Fn14 axis as a therapeutic target for post-angioplasty restenosis.","date":"2019","source":"EBioMedicine","url":"https://pubmed.ncbi.nlm.nih.gov/31395500","citation_count":30,"is_preprint":false},{"pmid":"19513634","id":"PMC_19513634","title":"Therapeutic targeting of TWEAK/Fnl4 in cancer: exploiting the intrinsic tumor cell killing capacity of the pathway.","date":"2009","source":"Results and problems in cell differentiation","url":"https://pubmed.ncbi.nlm.nih.gov/19513634","citation_count":29,"is_preprint":false},{"pmid":"23816995","id":"PMC_23816995","title":"Increased expression of atrogenes and TWEAK family members after severe burn injury in nonburned human skeletal muscle.","date":"2013","source":"Journal of burn care & research : official publication of the American Burn Association","url":"https://pubmed.ncbi.nlm.nih.gov/23816995","citation_count":29,"is_preprint":false},{"pmid":"17187350","id":"PMC_17187350","title":"How to tweak a beak: molecular techniques for studying the evolution of size and shape in Darwin's finches and other birds.","date":"2007","source":"BioEssays : news and reviews in molecular, cellular and developmental biology","url":"https://pubmed.ncbi.nlm.nih.gov/17187350","citation_count":29,"is_preprint":false},{"pmid":"14555217","id":"PMC_14555217","title":"TWE-PRIL; a fusion protein of TWEAK and APRIL.","date":"2003","source":"Biochemical pharmacology","url":"https://pubmed.ncbi.nlm.nih.gov/14555217","citation_count":28,"is_preprint":false},{"pmid":"24337061","id":"PMC_24337061","title":"TWEAK/Fn14 signaling mediates gastric cancer cell resistance to 5-fluorouracil via NF-κB activation.","date":"2013","source":"International journal of oncology","url":"https://pubmed.ncbi.nlm.nih.gov/24337061","citation_count":28,"is_preprint":false},{"pmid":"26499979","id":"PMC_26499979","title":"Serum levels of TWEAK in patients with psoriasis vulgaris.","date":"2015","source":"Cytokine","url":"https://pubmed.ncbi.nlm.nih.gov/26499979","citation_count":27,"is_preprint":false},{"pmid":"32200423","id":"PMC_32200423","title":"Controversies in TWEAK-Fn14 signaling in skeletal muscle atrophy and regeneration.","date":"2020","source":"Cellular and molecular life sciences : CMLS","url":"https://pubmed.ncbi.nlm.nih.gov/32200423","citation_count":26,"is_preprint":false},{"pmid":"29143982","id":"PMC_29143982","title":"TWEAK/Fn14 interaction induces proliferation and migration in human airway smooth muscle cells via activating the NF-κB pathway.","date":"2018","source":"Journal of cellular biochemistry","url":"https://pubmed.ncbi.nlm.nih.gov/29143982","citation_count":26,"is_preprint":false},{"pmid":"33512736","id":"PMC_33512736","title":"TWEAK-Fn14 as a common pathway in the heart and the kidneys in cardiorenal syndrome.","date":"2021","source":"The Journal of pathology","url":"https://pubmed.ncbi.nlm.nih.gov/33512736","citation_count":24,"is_preprint":false},{"pmid":"31097613","id":"PMC_31097613","title":"TRAIL, OPG, and TWEAK in kidney disease: biomarkers or therapeutic targets?","date":"2019","source":"Clinical science (London, England : 1979)","url":"https://pubmed.ncbi.nlm.nih.gov/31097613","citation_count":24,"is_preprint":false},{"pmid":"29971943","id":"PMC_29971943","title":"TWEAK/Fn14 mediates atrial-derived HL-1 myocytes hypertrophy via JAK2/STAT3 signalling pathway.","date":"2018","source":"Journal of cellular and molecular medicine","url":"https://pubmed.ncbi.nlm.nih.gov/29971943","citation_count":24,"is_preprint":false},{"pmid":"17485258","id":"PMC_17485258","title":"Tweak and FN14 in central nervous system health and disease.","date":"2007","source":"Frontiers in bioscience : a journal and virtual library","url":"https://pubmed.ncbi.nlm.nih.gov/17485258","citation_count":23,"is_preprint":false},{"pmid":"31051431","id":"PMC_31051431","title":"VPS29, a tweak tool of endosomal recycling.","date":"2019","source":"Current opinion in cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/31051431","citation_count":23,"is_preprint":false},{"pmid":"24406502","id":"PMC_24406502","title":"Nrf2 protects against TWEAK-mediated skeletal muscle wasting.","date":"2014","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/24406502","citation_count":23,"is_preprint":false},{"pmid":"22977477","id":"PMC_22977477","title":"Therapeutic potential of the TWEAK/Fn14 pathway in intractable gastrointestinal cancer.","date":"2010","source":"Experimental and therapeutic medicine","url":"https://pubmed.ncbi.nlm.nih.gov/22977477","citation_count":23,"is_preprint":false},{"pmid":"22649723","id":"PMC_22649723","title":"A Bioinformatics Resource for TWEAK-Fn14 Signaling Pathway.","date":"2012","source":"Journal of signal transduction","url":"https://pubmed.ncbi.nlm.nih.gov/22649723","citation_count":23,"is_preprint":false},{"pmid":"36626628","id":"PMC_36626628","title":"TWEAK/FN14 promotes profibrogenic pathway activation in Prominin-1-expressing hepatic progenitor cells in biliary atresia.","date":"2023","source":"Hepatology (Baltimore, Md.)","url":"https://pubmed.ncbi.nlm.nih.gov/36626628","citation_count":21,"is_preprint":false},{"pmid":"34246833","id":"PMC_34246833","title":"Bacterial effectors mimicking ubiquitin-proteasome pathway tweak plant immunity.","date":"2021","source":"Microbiological research","url":"https://pubmed.ncbi.nlm.nih.gov/34246833","citation_count":21,"is_preprint":false},{"pmid":"28025386","id":"PMC_28025386","title":"Tweak up-regulates endothelin-1 system in mouse and human endothelial cells.","date":"2016","source":"Cardiovascular research","url":"https://pubmed.ncbi.nlm.nih.gov/28025386","citation_count":21,"is_preprint":false},{"pmid":"30414907","id":"PMC_30414907","title":"TWEAK/Fn14 Interaction Confers Aggressive Properties to Cutaneous Squamous Cell Carcinoma.","date":"2018","source":"The Journal of investigative dermatology","url":"https://pubmed.ncbi.nlm.nih.gov/30414907","citation_count":20,"is_preprint":false},{"pmid":"29266762","id":"PMC_29266762","title":"The TWEAK/Fn14 pathway is required for calcineurin inhibitor toxicity of the kidneys.","date":"2018","source":"American journal of transplantation : official journal of the American Society of Transplantation and the American Society of Transplant Surgeons","url":"https://pubmed.ncbi.nlm.nih.gov/29266762","citation_count":20,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":49031,"output_tokens":8487,"usd":0.137199},"stage2":{"model":"claude-opus-4-6","input_tokens":12558,"output_tokens":4576,"usd":0.265785},"total_usd":0.402984,"stage1_batch_id":"msgbatch_01RPbVc9PDH9zQmYK5Hs6srm","stage2_batch_id":"msgbatch_01JVbVLzeKN2yVrsGp3zC8aM","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2001,\n      \"finding\": \"TWEAK (TNFSF12) binds to TweakR/Fn14, a small TNF receptor family member cloned from HUVEC library, with an affinity constant Kd of 2.3 ± 0.1 nM; the TweakR cytoplasmic domain binds TRAF1, TRAF2, and TRAF3; cross-linking of TweakR induces HUVEC growth; soluble TweakR inhibits endothelial cell migration and corneal angiogenesis in vivo.\",\n      \"method\": \"Five binding assays including receptor cloning, affinity measurement, Co-IP/pulldown for TRAF binding, HUVEC growth assay, corneal angiogenesis model\",\n      \"journal\": \"Immunity\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 — receptor cloning with multiple orthogonal binding assays, TRAF pulldown, and in vivo functional validation in one study\",\n      \"pmids\": [\"11728344\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2003,\n      \"finding\": \"TWEAK sequentially activates two NF-κB cascades: (1) rapid IκBα phosphorylation generating p50/RelA complexes via canonical IKK complex (NEMO-dependent); (2) prolonged activation via proteasome-mediated NF-κB2/p100 processing generating RelB complexes (NEMO-independent, requiring IKK1 and NF-κB-inducing kinase). Fn14 with mutated TRAF-binding site cannot activate NF-κB; cells deficient in TRAF2 and TRAF5 fail to show p100 processing and IκBα phosphorylation.\",\n      \"method\": \"Expression cloning, IκBα phosphorylation assays, EMSA, TRAF-binding site mutagenesis, TRAF2/TRAF5 knockout cells, proteasome inhibitor studies\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 — multiple orthogonal methods including mutagenesis, KO cells, and biochemical assays in one study; foundational mechanism paper\",\n      \"pmids\": [\"12840022\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2002,\n      \"finding\": \"TWEAK-induced cell death occurs through multiple pathways: caspase-8 and caspase-3-dependent apoptosis in HSC3 cells; cathepsin B-dependent necrosis (with lysosomal cathepsin B release to cytosol) in IFN-γ-treated HT-29 cells; and indirect TNF-α-mediated cell death in Kym-1 cells. DR3 is not the relevant receptor as TWEAK-sensitive cell lines do not express DR3 but bind TWEAK via a distinct receptor.\",\n      \"method\": \"Caspase activity assays, pan-caspase inhibitors, cathepsin B inhibitors, lysosomal fractionation, anti-TNF-α blocking antibodies, DR3 expression analysis by Western blot and RT-PCR\",\n      \"journal\": \"Journal of immunology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 — multiple cell lines with pharmacological inhibitors and mechanistic dissection of distinct death pathways\",\n      \"pmids\": [\"11777967\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2000,\n      \"finding\": \"TWEAK is expressed on the surface of IFN-γ-stimulated monocytes (but not IFN-α or LPS-stimulated) and mediates cytotoxic killing of tumor cells; anti-TWEAK mAb partially inhibited monocyte cytotoxicity, with near-complete inhibition in combination with anti-TRAIL.\",\n      \"method\": \"Anti-human TWEAK mAb generation, flow cytometry, cytotoxicity assays with neutralizing antibodies\",\n      \"journal\": \"The Journal of experimental medicine\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — reciprocal antibody blocking with clear functional readout, replicated finding\",\n      \"pmids\": [\"11067885\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2000,\n      \"finding\": \"TWEAK and DR3/WSL-1/TRAMP do not interact in in vitro binding assays; TWEAK binds to cells lacking DR3 surface expression and shows biological activity there; cells from DR3 knockout mice retain TWEAK-binding capacity, demonstrating DR3 is not the major TWEAK receptor.\",\n      \"method\": \"In vitro binding assay, flow cytometry, DR3 knockout mouse cells\",\n      \"journal\": \"FEBS letters\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 — direct binding assay and genetic KO cells providing clear negative result for DR3 as TWEAK receptor\",\n      \"pmids\": [\"11094155\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2002,\n      \"finding\": \"An endogenous hybrid mRNA (TWE-PRIL) encodes a fusion protein composed of TWEAK cytoplasmic and transmembrane domains fused to the APRIL C-terminal receptor-binding domain; TWE-PRIL is membrane-anchored, presents APRIL's receptor-binding domain at the cell surface, and is biologically active (stimulates cycling in T- and B-lymphoma cell lines). Detected in primary human T lymphocytes and monocytic cell lines by RACE, RT-PCR, cDNA library screening, RNase protection assay.\",\n      \"method\": \"RACE, RT-PCR, cDNA library screening, RNase protection assay, flow cytometry, proliferation assay\",\n      \"journal\": \"The EMBO journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 — multiple orthogonal methods confirming endogenous expression and biological activity of fusion protein\",\n      \"pmids\": [\"12411489\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2005,\n      \"finding\": \"TWEAK selectively stimulates liver progenitor (oval) cell proliferation via its receptor Fn14; transgenic TWEAK overexpression in hepatocytes causes oval cell hyperplasia; TWEAK-adenovirus induced oval cell expansion in WT but not Fn14-null mice; DDC-induced oval cell expansion was reduced in Fn14-null mice and by anti-TWEAK mAb; TWEAK stimulated proliferation of oval cell cultures directly.\",\n      \"method\": \"Transgenic overexpression, adenoviral delivery, Fn14 knockout mice, blocking anti-TWEAK mAb, DDC injury model, cell culture proliferation assays\",\n      \"journal\": \"The Journal of clinical investigation\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — genetic loss-of-function (Fn14 KO), pharmacological blockade, and gain-of-function (Tg/adenovirus) with concordant phenotypic readouts\",\n      \"pmids\": [\"16110324\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"TWEAK binds to mesenchymal progenitor cells (all lineages) and induces NF-κB activation, pro-survival, pro-proliferative and homing receptor gene expression. TWEAK promotes C2C12 myoblast proliferation and inhibits terminal differentiation. Fn14-deficient primary myoblasts show reduced proliferative capacity and altered myotube formation. After cardiotoxin injury, Fn14-deficient mice show reduced inflammatory response and delayed muscle fiber regeneration.\",\n      \"method\": \"Cell binding assays, NF-κB reporter assays, Fn14 knockout mice, C2C12 and primary myoblast proliferation/differentiation assays, cardiotoxin muscle injury model\",\n      \"journal\": \"The EMBO journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — genetic KO with multiple in vitro and in vivo functional readouts, replicated across cell types\",\n      \"pmids\": [\"17124496\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"TWEAK induces NF-κB activation in tubular epithelial cells (MCT line) via IκBα degradation and RelA nuclear translocation, leading to increased MCP-1, RANTES, and IL-6 expression. In vivo systemic TWEAK administration induces renal NF-κB activation, chemokine expression and interstitial inflammation; neutralization of TWEAK with blocking antibody reduces tubular chemokine expression and macrophage infiltration after folic acid-induced AKI. Parthenolide (IκBα degradation inhibitor) blocked all TWEAK-induced effects.\",\n      \"method\": \"In vitro NF-κB activation assays, Western blot for IκBα, nuclear translocation assays, ELISA, in vivo TWEAK administration in mice, anti-TWEAK mAb neutralization, folic acid AKI model\",\n      \"journal\": \"Journal of the American Society of Nephrology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — mechanistic pathway dissection with pharmacological inhibition and in vivo neutralizing antibody validation\",\n      \"pmids\": [\"18235096\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"Full-length human TWEAK is processed intracellularly by the serine protease furin at amino acid residues 90–93 (the predominant furin recognition site). Full-length, membrane-anchored TWEAK can bind Fn14 on neighboring cells (juxtacrine signaling) and activate the NF-κB signaling pathway.\",\n      \"method\": \"Furin inhibitor studies, site-directed mutagenesis of furin cleavage site, co-culture assays, NF-κB reporter assays\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — mutagenesis of active site combined with functional NF-κB activation assay, mechanistically defining furin cleavage and juxtacrine activity\",\n      \"pmids\": [\"20385556\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"TWEAK/Fn14 interaction in osteoblastic MC3T3-E1 cells induces RANTES via PI3K-Akt (not NF-κB); inhibits BMP-2-induced osteoblast differentiation markers via MAPK-ERK; and upregulates RANKL expression via MAPK-ERK pathway. All effects abolished by Fn14-Fc chimera decoy receptor.\",\n      \"method\": \"PI3K inhibitors, ERK inhibitors, MAPK pathway analysis, BMP-2 differentiation assays, RANKL expression measurement, Fn14-Fc neutralization\",\n      \"journal\": \"Arthritis research & therapy\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — pathway dissection with pharmacological inhibitors and decoy receptor blockade in single study\",\n      \"pmids\": [\"16945157\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"TWEAK and TNFα reduce renal Klotho expression through NF-κB-dependent mechanism. TWEAK promotes RelA binding to the Klotho promoter and induces histone deacetylation of the Klotho promoter. NF-κB inhibitor parthenolide and histone deacetylase inhibitor both prevent TWEAK-induced Klotho downregulation. Confirmed in vivo: TWEAK blockade or genetic absence abrogated injury-related decrease in renal Klotho.\",\n      \"method\": \"siRNA knockdown of IκBα, parthenolide NF-κB inhibition, HDAC inhibitor, chromatin immunoprecipitation (ChIP), TWEAK knockout mice, anti-TWEAK antibody in folic acid AKI model\",\n      \"journal\": \"Journal of the American Society of Nephrology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 — ChIP showing direct RelA promoter binding, multiple pharmacological inhibitors and genetic KO with concordant results\",\n      \"pmids\": [\"21719790\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"TWEAK activates the noncanonical NF-κB pathway (through p100 processing, RelB, IKKα, and NIK) to promote myoblast fusion into multinucleated myotubes. Loss of cIAP1 leads to constitutive noncanonical NF-κB activation and increased myotube nuclei. Low-dose TWEAK, which preferentially activates the noncanonical pathway, increases myoblast fusion without causing atrophy.\",\n      \"method\": \"cIAP1 knockout, siRNA knockdown of p100/RelB/IKKα/NIK/TRAF3, p52/RelB overexpression, NF-κB reporter assays, myotube fusion quantification\",\n      \"journal\": \"Science signaling\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 — multiple genetic perturbations (KO, siRNA, overexpression) with consistent phenotypic readouts establishing noncanonical NF-κB as the pathway for TWEAK-induced myoblast fusion\",\n      \"pmids\": [\"23074266\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"TWEAK transactivates EGFR in renal tubular epithelial cells via Fn14-dependent ADAM17 activation and subsequent release of EGFR ligands HB-EGF and TGFα. TWEAK-induced proinflammatory factor expression and ERK activation require EGFR signaling (blocked by erlotinib or ADAM17 inhibitor), while TWEAK-mediated NF-κB activation is EGFR-independent.\",\n      \"method\": \"EGFR phosphorylation assays, ADAM17 inhibitor (WTACE-2), EGFR kinase inhibitor (erlotinib), in vivo TWEAK administration in mice, HB-EGF/TGFα measurement\",\n      \"journal\": \"The Journal of pathology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 — pharmacological dissection of ADAM17-EGFR axis and in vivo validation, mechanistically separating EGFR-dependent ERK from EGFR-independent NF-κB pathways\",\n      \"pmids\": [\"24037740\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"CD163 functions as a decoy receptor for soluble TWEAK during ischemia; soluble CD163 sequesters TWEAK and inhibits TWEAK-induced canonical NF-κB and Notch signaling required for myogenic progenitor cell proliferation. CD163-knockout mice have transiently elevated TWEAK levels, stimulating muscle satellite cell proliferation and tissue regeneration.\",\n      \"method\": \"CD163 knockout mice, soluble TWEAK binding assays, NF-κB and Notch signaling assays, ischemic injury model, macrophage internalization experiments\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — genetic KO with mechanistic signaling readouts and in vivo ischemia model validation\",\n      \"pmids\": [\"26242746\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"CD163-expressing macrophages can bind and internalize TWEAK protein (demonstrated in vitro with exogenously added TWEAK from supernatants), suggesting CD163 is a scavenger receptor for TWEAK.\",\n      \"method\": \"In vitro TWEAK binding and internalization assay with CD163-expressing macrophages\",\n      \"journal\": \"Atherosclerosis\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 — single-method in vitro binding/internalization experiment\",\n      \"pmids\": [\"19473660\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"TWEAK/Fn14 signaling regulates skeletal muscle PGC-1α levels: TWEAK reduces PGC-1α and mitochondrial content (~50%) in skeletal muscle. PGC-1α levels are significantly increased in TWEAK-KO and Fn14-KO mice upon denervation. Transgenic PGC-1α overexpression inhibits muscle wasting in TWEAK-Tg mice, inhibits TWEAK-induced NF-κB activation (~50%), reduces atrogene (MAFbx, MuRF1) expression (~90%), and prevents Fn14 induction in denervated muscle.\",\n      \"method\": \"TWEAK-KO and Fn14-KO mice, TWEAK-Tg mice, PGC-1α Tg overexpression, denervation model, NF-κB activation assays, atrogene expression measurement, mitochondrial content quantification\",\n      \"journal\": \"FASEB journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — multiple genetic models (KO and Tg) with consistent mechanistic readouts\",\n      \"pmids\": [\"24327607\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"TWEAK decreases PGC-1α expression in tubular cells via NF-κB-dependent histone H3 deacetylation at the PGC-1α promoter, leading to decreased expression of PGC-1α target genes and reduced mitochondrial membrane potential. HDAC inhibitors and NF-κB inhibitors both prevent TWEAK-induced PGC-1α downregulation. Adenoviral PGC-1α overexpression rescues TWEAK-induced mitochondrial dysfunction.\",\n      \"method\": \"Chromatin immunoprecipitation (histone deacetylation at PGC-1α promoter), NF-κB inhibitors, HDAC inhibitors, adenoviral PGC-1α overexpression, mitochondrial membrane potential assays, neutralizing anti-TWEAK antibodies in folic acid AKI\",\n      \"journal\": \"Kidney international\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 — ChIP for mechanistic epigenetic regulation, multiple pharmacological inhibitors, in vivo neutralization, and rescue experiment\",\n      \"pmids\": [\"26535995\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"TWEAK/Fn14 activation drives a second wave of tubular cell death during AKI through necroptosis. In cultured tubular cells, TWEAK induces apoptosis in a proinflammatory environment; caspase inhibition switches this to necroptosis. In folic acid AKI mice, Fn14 ablation and RIPK1 inhibition (Nec-1) or RIPK3/MLKL deficiency protect against a late (72–96 h) wave of cell death and kidney dysfunction, but not early (48 h) cell death.\",\n      \"method\": \"Necrostatin-1 (RIPK1 inhibitor), genetic RIPK3 deficiency, MLKL deficiency, Fn14 genetic ablation, caspase inhibitors, folic acid AKI model, tubular cell culture with TWEAK treatment\",\n      \"journal\": \"Proceedings of the National Academy of Sciences\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — multiple genetic knockouts and pharmacological inhibitors with distinct temporal dissection of cell death pathways in vivo and in vitro\",\n      \"pmids\": [\"29588419\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"TRAF3IP2 mediates TWEAK/Fn14 pro-fibrotic signaling in cardiac fibroblasts: exogenous TWEAK upregulates TRAF3IP2 in a time- and dose-dependent manner (positive feedback), and TWEAK promotes TRAF3IP2 nuclear translocation. Silencing TRAF3IP2 inhibits TWEAK-induced p38 MAPK, NF-κB and AP-1 activation, inflammatory cytokine expression, MMP activation, collagen synthesis, and CF proliferation/migration. Genetic TRAF3IP2 ablation in vivo blocks TWEAK-induced cardiac fibrosis and dysfunction.\",\n      \"method\": \"TRAF3IP2 overexpression and siRNA knockdown in cardiac fibroblasts, p38 MAPK/NF-κB/AP-1 activation assays, collagen secretion assays, TWEAK infusion in mice, TRAF3IP2 knockout mice\",\n      \"journal\": \"Journal of molecular and cellular cardiology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — genetic KO in vivo and siRNA in vitro with multiple signaling pathway readouts demonstrating TRAF3IP2 as causal mediator\",\n      \"pmids\": [\"29981796\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2004,\n      \"finding\": \"A short variant of TWEAK (shortTWEAK) forms a complex with GSK-3β in a yeast two-hybrid system and colocalizes with GSK-3β in the nucleus of neuroblastoma cells. TWEAK is internalized and translocates to the nucleus, causing IκBα degradation, nuclear translocation of both GSK-3β and p65, and NF-κB-driven gene expression. LiCl (GSK-3β inhibitor) counteracts TWEAK-induced IL-8 expression.\",\n      \"method\": \"Yeast two-hybrid, colocalization by immunofluorescence, TWEAK internalization assay, NF-κB reporter assays, LiCl inhibition of GSK-3β\",\n      \"journal\": \"FEBS letters\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2–3 — yeast two-hybrid with functional pharmacological validation, but nuclear TWEAK mechanism not extensively replicated\",\n      \"pmids\": [\"15147869\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"TWEAK/Fn14 signaling drives VSMC proliferation and migration through upregulation of cyclins D1, CDK4/CDK6, and downregulation of CDK inhibitor p15INK4B via ERK and Akt activation. Genetic depletion of Tnfrsf12a or Tnfsf12 and pharmacological anti-TWEAK antibody reduce neointimal formation, cell proliferation, cyclin D1/CDK4/6, and increase p15INK4B in wire-injured mouse femoral arteries.\",\n      \"method\": \"Next-generation sequencing of TWEAK-regulated genes, Tnfrsf12a/Tnfsf12 knockout VSMCs, wound-healing/Transwell migration assays, cyclin/CDK expression by Western blot, wire injury mouse model, anti-TWEAK mAb treatment, ERK/Akt inhibition\",\n      \"journal\": \"EBioMedicine\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — genetic KO in vitro and in vivo with mechanistic cell cycle pathway dissection and pharmacological blockade\",\n      \"pmids\": [\"31395500\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"TWEAK/Fn14 activates NF-κB in cardiac fibroblasts, increasing MMP-9 production and promoting CF proliferation and collagen synthesis; siRNA knockdown of Fn14 or NF-κB inhibitor (PDTC) prevents these effects.\",\n      \"method\": \"Fn14 siRNA knockdown, PDTC NF-κB inhibition, MMP-9 activity assay, collagen synthesis measurement, CF proliferation assays\",\n      \"journal\": \"Molecular biology reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2–3 — siRNA with multiple functional readouts but single lab study\",\n      \"pmids\": [\"22555979\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"In keratinocytes, IFN-γ suppresses miR-149, and the TWEAK receptor Fn14 is a direct target of miR-149 (validated by 3'UTR reporter assays). IFN-γ-mediated miR-149 suppression amplifies keratinocyte inflammatory responses to TWEAK; synthetic miR-149 overexpression is protective in imiquimod-induced psoriasis mouse model.\",\n      \"method\": \"miR-149 target validation (reporter assays), miR-149 overexpression/inhibition in keratinocytes, intradermal miR-149 injection in mice, imiquimod psoriasis model\",\n      \"journal\": \"The Journal of allergy and clinical immunology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — direct target validation of Fn14 as miR-149 target with in vivo functional validation in psoriasis model\",\n      \"pmids\": [\"33705829\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"In keratinocytes, TWEAK/Fn14 signaling synergizes with TNF and IL-17A to upregulate psoriasis-signature genes (CXC chemokines, IL-23A, IL-36G, S100A8/9, SERPINB1/B9). Keratinocyte-specific deletion of Fn14 reduces imiquimod-induced skin inflammation, epidermal hyperplasia, and psoriasis gene expression.\",\n      \"method\": \"Keratinocyte-specific Fn14 knockout mice, transcriptomic analysis in human keratinocytes, anti-TWEAK therapeutic treatment, imiquimod psoriasis model, combination cytokine stimulation assays\",\n      \"journal\": \"Science immunology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — cell-type-specific genetic KO with transcriptomic mechanistic data and in vivo phenotypic validation\",\n      \"pmids\": [\"34797693\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"TWEAK/Fn14 signaling mediates HL-1 atrial myocyte hypertrophy through the JAK2/STAT3 pathway: TWEAK increases ANP and Troponin T expression, Fn14 knockdown blocks these effects, and siRNA inhibition of JAK2 or STAT3 attenuates TWEAK-induced hypertrophy.\",\n      \"method\": \"Fn14 knockdown, JAK2/STAT3 siRNA, hypertrophy markers (ANP, Troponin T) measurement in HL-1 atrial myocytes\",\n      \"journal\": \"Journal of cellular and molecular medicine\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2–3 — siRNA knockdown with hypertrophy markers but single lab study in a cell line\",\n      \"pmids\": [\"29971943\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"TWEAK upregulates ECE-1 (endothelin-converting enzyme-1) and ET-1 production in endothelial cells via AP-1 (through ERK1/2 and JNK) and NF-κB transcription factors acting on the ECE-1 promoter. Both AP-1 and NF-κB inhibitors partially block the effect. Validated in vivo: TWEAK injection in mice increases aortic and lung ECE-1 and ET-1 expression.\",\n      \"method\": \"ECE-1 promoter deletion/transfection assays, EMSA for AP-1 and NF-κB, ERK/JNK inhibitors, NF-κB inhibitor, in vivo TWEAK injection in mice, Western blot and qPCR, ET-1 ELISA\",\n      \"journal\": \"Cardiovascular research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — promoter dissection with EMSA and pharmacological inhibitors, confirmed in vivo\",\n      \"pmids\": [\"28025386\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"TWEAK secreted by Th17 cells promotes epithelial-mesenchymal transition (EMT) in colorectal cancer cells by binding to Fn14 on tumor cells, enhancing tumor migration and invasion. In mouse models, Fn14 knockout (CRISPR) or Tnfsf12 deletion reduces liver metastases and prolongs survival. Cotransfer of Th17 cells with tumor cells promotes liver metastasis.\",\n      \"method\": \"Single-cell RNA sequencing, CRISPR-mediated Fn14 knockout, Tnfsf12 knockout mice, LNP-encapsulated siRNA, Il17a knockout mice, Th17 cotransfer experiments, EMT markers\",\n      \"journal\": \"Cancer research\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — multiple genetic loss-of-function approaches with clear in vivo metastasis phenotype and mechanistic EMT readout\",\n      \"pmids\": [\"38335276\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"IL-13-induced apoptosis (caspase-3 activation) in intestinal epithelial explants requires TWEAK, Fn14, and endogenous TNF-α secreted via ADAM17. Conversely, TNF-α-induced caspase activation requires IL-13, TWEAK, and Fn14. This was demonstrated using intestinal explants from TWEAK knockout mice.\",\n      \"method\": \"TWEAK knockout mice intestinal explants, ADAM17 inhibition, caspase-3 activation assays, IL-13 and TNF-α treatments\",\n      \"journal\": \"Gastroenterology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — genetic KO with mechanistic dissection of cytokine crosstalk and identified ADAM17-dependent pathway\",\n      \"pmids\": [\"21893119\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"TWEAK administration induces cystogenesis and worsens the ADPKD phenotype; anti-TWEAK antibodies slow cystic progression and preserve renal function. The anti-cystogenic effect is associated with decreased MAPK signaling (cell proliferation-related), decreased NF-κB pathway activation, reduced fibrosis and apoptosis, and indirect decrease of macrophage recruitment.\",\n      \"method\": \"TWEAK administration in murine ADPKD model, anti-TWEAK antibody treatment, MAPK/NF-κB signaling assays, macrophage infiltration quantification, survival analysis\",\n      \"journal\": \"Journal of the American Society of Nephrology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2–3 — pharmacological gain- and loss-of-function in vivo with pathway readouts but no genetic KO confirmation\",\n      \"pmids\": [\"34155062\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"TNFSF12 (TWEAK) is a TNF superfamily cytokine that exists as membrane-anchored (processed intracellularly by furin at residues 90–93) and soluble isoforms; it binds its primary receptor Fn14 (TNFRSF12A) with ~2.3 nM affinity, recruiting TRAF1/2/3 to activate both canonical (rapid IκBα phosphorylation, RelA/p50) and noncanonical (IKK1/NIK-dependent p100 processing, RelB) NF-κB pathways; additionally, TWEAK transactivates EGFR via ADAM17-mediated shedding of HB-EGF/TGFα to drive ERK-dependent inflammation, upregulates ECE-1/ET-1 via AP-1 and NF-κB, and can signal through GSK-3β in the nucleus; soluble TWEAK is scavenged by CD163 acting as a decoy receptor; cell-type-specific responses include promotion of progenitor/myoblast proliferation via noncanonical NF-κB, induction of cathepsin B-dependent necrosis or caspase-dependent apoptosis, RIPK1/RIPK3/MLKL-dependent necroptosis in renal tubular cells, epigenetic downregulation of Klotho and PGC-1α via RelA-driven histone deacetylation, and promotion of EMT in tumor cells, collectively placing TWEAK/Fn14 as a multifunctional inflammatory axis regulating tissue repair, muscle biology, renal injury, and cancer progression.\"\n}\n```","stage2_raw":"```json\n{\n  \"mechanistic_narrative\": \"TNFSF12 (TWEAK) is a TNF superfamily cytokine that functions as a pleiotropic regulator of inflammation, tissue repair, cell death, and proliferation through engagement of its receptor Fn14 (TNFRSF12A). TWEAK exists as a membrane-anchored form processed intracellularly by furin at residues 90–93 and as a soluble form scavenged by the decoy receptor CD163; membrane TWEAK signals in a juxtacrine manner, while soluble TWEAK acts at a distance, with both forms activating canonical (IκBα/RelA/p50) and noncanonical (NIK/IKK1/p100→p52/RelB) NF-κB pathways through TRAF1/2/3 recruitment to the Fn14 cytoplasmic domain [PMID:11728344, PMID:12840022, PMID:20385556, PMID:26242746]. Beyond NF-κB, TWEAK/Fn14 transactivates EGFR via ADAM17-mediated shedding of HB-EGF and TGFα to drive ERK-dependent inflammatory gene expression, activates PI3K–Akt and MAPK–ERK cascades regulating cell cycle progression, and epigenetically silences protective genes including Klotho and PGC-1α through RelA-directed histone deacetylation at their promoters [PMID:24037740, PMID:21719790, PMID:26535995, PMID:31395500]. These signaling outputs underlie context-dependent biological activities: stimulation of progenitor and myoblast proliferation and fusion via noncanonical NF-κB, induction of cathepsin B–dependent necrosis or caspase-dependent apoptosis, RIPK1/RIPK3/MLKL-dependent necroptosis in renal tubular cells during acute kidney injury, promotion of epithelial–mesenchymal transition driving colorectal cancer metastasis, and pro-fibrotic remodeling in cardiac tissue through the adaptor TRAF3IP2 [PMID:23074266, PMID:11777967, PMID:29588419, PMID:38335276, PMID:29981796].\",\n  \"teleology\": [\n    {\n      \"year\": 2000,\n      \"claim\": \"Establishing that TWEAK is a monocyte-surface cytokine and ruling out DR3 as its receptor resolved a receptor-identity question that had complicated the field since TWEAK's cloning.\",\n      \"evidence\": \"Flow cytometry on IFN-γ-stimulated monocytes with anti-TWEAK mAb blocking; DR3-KO mouse cells retaining TWEAK binding\",\n      \"pmids\": [\"11067885\", \"11094155\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Identity of the true TWEAK receptor was still unknown\", \"Signaling pathways downstream of TWEAK binding uncharacterized\"]\n    },\n    {\n      \"year\": 2001,\n      \"claim\": \"Identification of Fn14 as the bona fide TWEAK receptor with 2.3 nM affinity and TRAF1/2/3 recruitment established the ligand–receptor pair and immediately linked it to angiogenesis and endothelial growth.\",\n      \"evidence\": \"Receptor cloning from HUVEC library, saturation binding, Co-IP/pulldown for TRAF binding, corneal angiogenesis model\",\n      \"pmids\": [\"11728344\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Which NF-κB pathways are activated downstream of TRAF recruitment was unresolved\", \"Relative roles of membrane vs. soluble TWEAK unknown\"]\n    },\n    {\n      \"year\": 2002,\n      \"claim\": \"Dissection of TWEAK-induced cell death revealed that it operates through at least three distinct mechanisms — caspase-dependent apoptosis, cathepsin B-dependent necrosis, and indirect TNF-α-mediated killing — establishing TWEAK as a context-dependent death ligand.\",\n      \"evidence\": \"Caspase/cathepsin B inhibitors, lysosomal fractionation, anti-TNF-α blocking in multiple cell lines\",\n      \"pmids\": [\"11777967\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Receptor mediating TWEAK death signaling not yet confirmed as Fn14\", \"Necroptotic pathway not yet explored\"]\n    },\n    {\n      \"year\": 2003,\n      \"claim\": \"Demonstration that TWEAK/Fn14 sequentially activates both canonical (NEMO-dependent, rapid IκBα phosphorylation) and noncanonical (IKK1/NIK-dependent p100 processing) NF-κB defined the dual signaling architecture downstream of Fn14.\",\n      \"evidence\": \"TRAF-binding site mutagenesis, TRAF2/TRAF5-KO cells, proteasome inhibitor studies, EMSA\",\n      \"pmids\": [\"12840022\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Cell-type-specific dominance of canonical vs. noncanonical pathways undetermined\", \"Non-NF-κB signaling arms not yet mapped\"]\n    },\n    {\n      \"year\": 2005,\n      \"claim\": \"Showing that TWEAK/Fn14 selectively drives liver progenitor (oval) cell proliferation in vivo provided the first genetic evidence that TWEAK acts as a tissue-repair mitogen for progenitor populations.\",\n      \"evidence\": \"Fn14-KO mice, TWEAK-transgenic and adenoviral overexpression, anti-TWEAK mAb, DDC injury model\",\n      \"pmids\": [\"16110324\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Downstream signaling pathway mediating oval cell proliferation not fully resolved\", \"Whether TWEAK drives other stem/progenitor compartments unknown\"]\n    },\n    {\n      \"year\": 2006,\n      \"claim\": \"Extension to myoblasts and osteoblasts revealed that TWEAK/Fn14 broadly controls mesenchymal progenitor proliferation and differentiation, engaging PI3K–Akt and MAPK–ERK in addition to NF-κB depending on cell type.\",\n      \"evidence\": \"Fn14-KO primary myoblasts, C2C12 differentiation, cardiotoxin injury model, PI3K/ERK inhibitors in osteoblasts\",\n      \"pmids\": [\"17124496\", \"16945157\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Mechanism by which TWEAK inhibits osteoblast differentiation vs. promoting myoblast proliferation not reconciled\", \"Relative contribution of canonical vs. noncanonical NF-κB to myoblast phenotype unclear\"]\n    },\n    {\n      \"year\": 2008,\n      \"claim\": \"In vivo renal studies established that TWEAK/Fn14 drives tubular NF-κB activation, chemokine production, and macrophage infiltration, positioning the axis as a central mediator of kidney inflammatory injury.\",\n      \"evidence\": \"Systemic TWEAK administration in mice, anti-TWEAK mAb in folic acid AKI, parthenolide NF-κB inhibition\",\n      \"pmids\": [\"18235096\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Direct vs. indirect effects on macrophage recruitment not separated\", \"Whether non-NF-κB pathways contribute to renal injury not explored\"]\n    },\n    {\n      \"year\": 2010,\n      \"claim\": \"Identification of furin cleavage at residues 90–93 and demonstration that uncleaved membrane TWEAK signals in trans via juxtacrine Fn14 engagement resolved how TWEAK exists in two functional forms.\",\n      \"evidence\": \"Furin cleavage-site mutagenesis, furin inhibitor, co-culture NF-κB reporter assays\",\n      \"pmids\": [\"20385556\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether other proteases contribute to shedding in specific tissues unknown\", \"Relative potency of membrane vs. soluble TWEAK in vivo not quantified\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Discovery that TWEAK epigenetically silences renal Klotho via RelA binding and histone deacetylation at its promoter revealed a chromatin-level mechanism by which TWEAK accelerates kidney aging/injury phenotypes.\",\n      \"evidence\": \"ChIP showing RelA at Klotho promoter, HDAC inhibitor rescue, TWEAK-KO mice, anti-TWEAK in folic acid AKI\",\n      \"pmids\": [\"21719790\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Which specific HDAC is recruited not identified\", \"Whether Klotho downregulation is the primary driver of renal pathology vs. a parallel effect unclear\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Selective genetic dissection showed that low-dose TWEAK promotes myoblast fusion specifically through the noncanonical NF-κB pathway (p100/RelB/IKKα/NIK), while high-dose TWEAK engages canonical NF-κB causing atrophy, resolving a dose-dependent signaling paradox.\",\n      \"evidence\": \"cIAP1-KO, siRNA of p100/RelB/IKKα/NIK/TRAF3, p52/RelB overexpression, myotube fusion assays\",\n      \"pmids\": [\"23074266\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"In vivo confirmation of dose-dependent pathway switching not performed\", \"Whether cIAP1 degradation is a physiological TWEAK-triggered event unclear\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Identification of ADAM17-mediated EGFR transactivation as an EGFR-dependent, NF-κB-independent arm of TWEAK signaling revealed a second major effector pathway that drives ERK-dependent inflammation through HB-EGF and TGFα shedding.\",\n      \"evidence\": \"ADAM17 inhibitor, erlotinib, EGFR phosphorylation assays, in vivo TWEAK administration\",\n      \"pmids\": [\"24037740\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether EGFR transactivation occurs in non-renal tissues unknown\", \"Direct mechanism linking Fn14 to ADAM17 activation not defined\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Establishing CD163 as a decoy/scavenger receptor for soluble TWEAK explained how macrophage phenotype regulates TWEAK bioavailability and muscle progenitor cell responses during ischemia.\",\n      \"evidence\": \"CD163-KO mice with elevated TWEAK and enhanced satellite cell proliferation, binding assays, ischemic injury model\",\n      \"pmids\": [\"26242746\", \"19473660\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether CD163 signals upon TWEAK binding or is purely a decoy not resolved\", \"Contribution of CD163-TWEAK axis in non-muscle tissues not explored\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Extension of the epigenetic silencing mechanism to PGC-1α — via NF-κB-driven histone deacetylation at its promoter — linked TWEAK to mitochondrial dysfunction in both muscle and kidney, unifying metabolic and inflammatory consequences of sustained TWEAK signaling.\",\n      \"evidence\": \"ChIP at PGC-1α promoter, HDAC/NF-κB inhibitors, adenoviral PGC-1α rescue, TWEAK-KO/Fn14-KO mice denervation model\",\n      \"pmids\": [\"26535995\", \"24327607\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Identity of the recruited HDAC enzyme not determined\", \"Whether TWEAK affects other metabolic gene promoters via the same mechanism unknown\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Demonstration that TWEAK/Fn14 triggers RIPK1/RIPK3/MLKL-dependent necroptosis as a temporally distinct late wave of tubular cell death in AKI established necroptosis as a TWEAK-mediated death modality and explained why caspase inhibition alone is insufficient for renal protection.\",\n      \"evidence\": \"Necrostatin-1, RIPK3-deficient and MLKL-deficient mice, Fn14-KO, folic acid AKI with temporal dissection\",\n      \"pmids\": [\"29588419\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether TWEAK directly activates RIPK1 or does so via TNF-α co-signaling not fully resolved\", \"Upstream signals determining apoptosis vs. necroptosis switching remain unclear\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Identification of TRAF3IP2 as a required adaptor for TWEAK-induced cardiac fibrosis — mediating p38 MAPK, NF-κB, and AP-1 activation — revealed a tissue-specific signaling node in fibroblasts.\",\n      \"evidence\": \"TRAF3IP2-KO mice, siRNA in cardiac fibroblasts, TWEAK infusion, collagen/MMP readouts\",\n      \"pmids\": [\"29981796\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How TRAF3IP2 is recruited to the Fn14 signaling complex not determined\", \"Whether TRAF3IP2 participates in TWEAK signaling in non-cardiac fibroblasts unknown\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Mechanistic dissection of VSMC proliferation showed TWEAK drives cell cycle progression via ERK/Akt-dependent cyclin D1–CDK4/6 upregulation and p15INK4B suppression, explaining TWEAK's pro-neointimal role after vascular injury.\",\n      \"evidence\": \"Tnfrsf12a/Tnfsf12-KO VSMCs, wire injury model, anti-TWEAK mAb, ERK/Akt inhibitors, NGS\",\n      \"pmids\": [\"31395500\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether noncanonical NF-κB contributes to VSMC phenotype not tested\", \"Relevance to human restenosis not established\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"TWEAK/Fn14 was shown to synergize with TNF and IL-17A in keratinocytes to amplify psoriasis-signature gene expression, and keratinocyte-specific Fn14 deletion protected against psoriasiform inflammation, establishing TWEAK as a co-driver of skin autoimmunity.\",\n      \"evidence\": \"Keratinocyte-specific Fn14-KO mice, imiquimod model, transcriptomics, miR-149 target validation for Fn14\",\n      \"pmids\": [\"34797693\", \"33705829\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether TWEAK blockade is therapeutically effective in human psoriasis untested\", \"Mechanism of TWEAK synergy with IL-17A at the promoter level not defined\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Th17-derived TWEAK was identified as a driver of colorectal cancer liver metastasis through Fn14-dependent EMT, demonstrating a tumor-extrinsic immune source of TWEAK that promotes cancer dissemination.\",\n      \"evidence\": \"scRNA-seq, CRISPR Fn14-KO tumor cells, Tnfsf12-KO mice, Th17 cotransfer experiments, EMT marker analysis\",\n      \"pmids\": [\"38335276\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether TWEAK-induced EMT is reversible upon Fn14 blockade not addressed\", \"Applicability beyond colorectal cancer not explored\", \"Contribution of other Th17 cytokines confounding in cotransfer experiments\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"Major unresolved questions include the structural basis of Fn14 signaling complex assembly, the identity of HDACs recruited to TWEAK-silenced promoters, the molecular switch determining whether TWEAK triggers apoptosis, necroptosis, or proliferation in a given cell, and the therapeutic window for TWEAK/Fn14 blockade in human disease.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"No crystal structure of TWEAK–Fn14 signaling complex available\", \"HDAC identity at Klotho and PGC-1α promoters not determined\", \"Dose- and context-dependent pathway switching mechanism not molecularly defined\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0048018\", \"supporting_discovery_ids\": [0, 3, 6, 7, 8, 27]},\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [1, 12, 13]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [3, 9]},\n      {\"term_id\": \"GO:0005576\", \"supporting_discovery_ids\": [9, 14]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"GO:0162582\", \"supporting_discovery_ids\": [0, 1, 8, 10, 13, 19, 21]},\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [3, 8, 24]},\n      {\"term_id\": \"R-HSA-5357801\", \"supporting_discovery_ids\": [2, 18, 28]},\n      {\"term_id\": \"R-HSA-1643685\", \"supporting_discovery_ids\": [27, 29]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"TNFRSF12A\", \"TRAF1\", \"TRAF2\", \"TRAF3\", \"CD163\", \"TRAF3IP2\", \"ADAM17\", \"GSK3B\"],\n    \"other_free_text\": []\n  }\n}\n```"}