{"gene":"WNT3A","run_date":"2026-04-28T23:00:23","timeline":{"discoveries":[{"year":1999,"finding":"Genetic epistasis shows that LEF-1 and TCF-1 transcription factors act downstream of WNT3A; double knockout of Lef1 and Tcf1 phenocopies Wnt3a-deficient mice (paraxial mesoderm defects, extra neural tubes, limb bud failure), demonstrating that LEF-1/TCF-1 are the primary nuclear effectors of WNT3A signaling in early mouse embryogenesis.","method":"Genetic double-knockout mouse (Lef1−/−Tcf1−/−) phenotypic analysis and epistasis","journal":"Genes & Development","confidence":"High","confidence_rationale":"Tier 2 — reciprocal genetic epistasis in vivo; strong phenocopy evidence; replicated by independent labs studying Wnt3a","pmids":["10090727"],"is_preprint":false},{"year":1998,"finding":"Soluble WNT3A secreted from transfected L cells stabilizes β-catenin in recipient cells and reorganizes the actin cytoskeleton (directional alignment of stress fibers, redistribution of villin to leading edges) in C57MG mammary epithelial cells, establishing that WNT3A acts as a diffusible paracrine signal that both activates β-catenin and remodels cytoskeletal architecture.","method":"Conditioned medium transfer assay, immunostaining for actin/adhesion proteins, β-catenin Western blot","journal":"Genes to Cells","confidence":"High","confidence_rationale":"Tier 2 — multiple orthogonal readouts (β-catenin stabilization, cytoskeletal reorganization) in defined cell-based assay; foundational paper","pmids":["9893023"],"is_preprint":false},{"year":2004,"finding":"WNT3A activates cell proliferation via two parallel pathways: (1) the canonical β-catenin/TCF-4 pathway and (2) a β-catenin-independent Raf-1–MEK–ERK cascade; ERK activation by WNT3A persists even when β-catenin is depleted by siRNA, and β-catenin can secondarily activate ERK through a TCF-4-dependent transcriptional event.","method":"siRNA knockdown of β-catenin and ERK, MEK inhibitor (U0126), dominant-negative TCF-4 transfection, proliferation and cell cycle assays in NIH3T3 cells","journal":"Journal of Cell Science","confidence":"High","confidence_rationale":"Tier 2 — multiple orthogonal genetic and pharmacological perturbations; two independent pathway arms defined","pmids":["15615777"],"is_preprint":false},{"year":2010,"finding":"WNT3A secretion requires PORCN-dependent lipid (palmitoyleoyl) modification at Ser209 of WNT3A, and the WNT3A–WLS complex accumulates when vacuolar acidification is inhibited; WLS is ER-resident in human cells and binds WNT3A through its lipid-binding domain, with vacuolar acidification needed to release palmitoylated WNT3A from WLS into secretory vesicles.","method":"V-ATPase inhibitor treatment, mutagenesis of Ser209, co-immunoprecipitation of WNT3A–WLS, cell fractionation, Xenopus convergent extension assay, structural modeling","journal":"Journal of Cell Science","confidence":"High","confidence_rationale":"Tier 1 — mutagenesis of lipidation site combined with pharmacological dissection and co-IP; multiple orthogonal methods","pmids":["20826466"],"is_preprint":false},{"year":2005,"finding":"WNT3A acts as a long-range signaling molecule from the primitive streak/dorsal posterior node that activates the Delta/Notch pathway to regulate perinodal Nodal expression (left-right determination) while simultaneously controlling the segmentation clock oscillations of both Wnt/β-catenin and Notch pathways, genetically linking the segmentation clock to left-right axis specification.","method":"Wnt3a knockout mouse analysis, in situ hybridization, genetic epistasis with Notch/Delta pathway components","journal":"Development","confidence":"High","confidence_rationale":"Tier 2 — in vivo knockout with defined molecular pathway placement; multiple target genes assessed","pmids":["16291790"],"is_preprint":false},{"year":2006,"finding":"Integrin-linked kinase (ILK) activity is required for WNT3A-induced β-catenin stabilization and nuclear translocation; ILK inhibition suppresses GSK-3β Ser9 phosphorylation triggered by WNT3A conditioned medium and reverses β-catenin/LEF-mediated transcription; ILK co-immunoprecipitates with APC and GSK-3β as part of a Wnt pathway complex.","method":"Pharmacological ILK inhibition (QLT-0267), molecular inhibition constructs, purified WNT3A protein treatment, co-IP of ILK with APC/GSK-3β, β-catenin stabilization and nuclear translocation assays","journal":"Oncogene","confidence":"High","confidence_rationale":"Tier 2 — reciprocal co-IP, pharmacological and molecular inhibition with multiple readouts; purified protein used to exclude secondary effects","pmids":["16799642"],"is_preprint":false},{"year":2009,"finding":"WNT3A stimulation, via Frizzled and Dishevelled, activates PI4-kinase type IIα to produce PtdIns(4)P; Dvl directly interacts with and activates PI4KIIα (increasing its Vmax for ATP and PtdIns), and Dvl, PI4KIIα, and PIP5KI form a ternary complex upon WNT3A stimulation to efficiently produce PtdIns(4,5)P2 from PtdIns.","method":"Co-immunoprecipitation, in vitro kinase assay with purified components, lipid measurement in HEK293T cells, dominant-negative Fz and Dvl constructs","journal":"Journal of Biological Chemistry","confidence":"High","confidence_rationale":"Tier 1 — direct binding and in vitro kinase activation assay combined with cellular co-IP; ternary complex reconstituted","pmids":["19561074"],"is_preprint":false},{"year":2009,"finding":"WNT3A-stimulated RhoA GTPase activity is required for full induction of a subset of canonical WNT3A target genes in a β-catenin-dependent transcriptional program, but RhoA does not affect β-catenin stabilization or nuclear translocation itself; Rho activation is an independent co-requirement for WNT3A-driven osteoblastic differentiation.","method":"RhoA inhibition (C3 toxin, dominant-negative RhoA), microarray mRNA expression, real-time RT-PCR, C3H10T1/2 osteoblastic differentiation assay","journal":"Cellular Signalling","confidence":"Medium","confidence_rationale":"Tier 2 — genome-wide transcriptomics plus pharmacological epistasis; single lab","pmids":["19482078"],"is_preprint":false},{"year":2011,"finding":"WNT3A stimulation of mammalian Frizzled receptors triggers guanine-nucleotide exchange on heterotrimeric Go/Gi proteins; this response is blocked by pertussis toxin (which ADP-ribosylates Go/Gi) and by a Wnt antagonist, demonstrating that mammalian Frizzled receptors function as bona fide GPCRs coupling to Go/i upon WNT3A binding.","method":"GTPγS-binding assay on rat brain membranes and cultured cells, pertussis toxin treatment, Wnt antagonist blockade","journal":"Biochemical Journal","confidence":"High","confidence_rationale":"Tier 1 — direct biochemical GEF assay on native membranes; pertussis toxin provides mechanistic specificity; multiple cellular contexts tested","pmids":["21128903"],"is_preprint":false},{"year":2013,"finding":"WNT3A undergoes basolateral secretion in polarized epithelial cells via a clathrin/adaptor protein-1 (AP-1)-dependent route, and this requires WLS recycling by AP-2; WNT3A carries two high-mannose-type N-glycans (at Asn87 and Asn298), and glycosylation processing at these sites does not redirect WNT3A to apical secretion (unlike WNT11, which uses complex-type glycan at Asn40 for apical targeting).","method":"Mass spectrometric glycan analysis, clathrin/AP-1 knockdown, AP-2 disruption, polarized MDCK cell secretion assay, WLS localization by immunostaining","journal":"Journal of Cell Science","confidence":"High","confidence_rationale":"Tier 1 — mass spectrometry for glycan identification combined with genetic knockdown of trafficking machinery; orthogonal readouts","pmids":["23613470"],"is_preprint":false},{"year":2006,"finding":"WNT3A activates the PI3K–Akt pathway independently of β-catenin in NIH3T3 fibroblasts; Wnt3a-induced Akt activation is abrogated by PI3K inhibitors (LY294002, wortmannin) but not by MEK inhibitor U0126, and activated Akt transiently accumulates in the nucleus; PI3K–Akt activation contributes to WNT3A-induced cell proliferation.","method":"PI3K inhibitors, Akt siRNA, β-catenin siRNA, subcellular fractionation/nuclear Akt detection, proliferation assay in NIH3T3 cells","journal":"Cellular Signalling","confidence":"Medium","confidence_rationale":"Tier 2 — pharmacological and siRNA epistasis; single lab but multiple perturbations","pmids":["17011750"],"is_preprint":false},{"year":2016,"finding":"WNT3A induces GSK-3β Ser9 phosphorylation and β-catenin accumulation through activation of the RhoA/ROCK axis; shRNA knockdown of RhoA or ROCK inhibition (Y27632) abolishes both GSK-3β phosphorylation and β-catenin accumulation induced by WNT3A; purified active ROCK directly phosphorylates recombinant GSK-3β in vitro.","method":"shRNA against RhoA, ROCK inhibitor Y27632, Tat-C3 (RhoA inhibitor), in vitro kinase assay with purified ROCK and recombinant GSK-3β, Western blot in RAW264.7 and HEK293 cells","journal":"Journal of Cellular Physiology","confidence":"High","confidence_rationale":"Tier 1 — in vitro kinase reconstitution of ROCK→GSK-3β plus genetic (shRNA) and pharmacological epistasis","pmids":["27575935"],"is_preprint":false},{"year":2018,"finding":"WNT3A promotes superoxide generation leading to Src-mediated Tyr42 phosphorylation of RhoA; phospho-Tyr42-RhoA binds to β-catenin via β-catenin's N-terminal domain and the complex translocates to the nucleus, where it associates with the Vimentin (Vim) gene promoter to drive vimentin transcription.","method":"Co-immunoprecipitation of p-Tyr42 RhoA with β-catenin, ChIP on Vim promoter, domain mapping (N-terminal β-catenin deletion), ROS detection, NADPH oxidase inhibition, ROCK2 inhibition","journal":"Redox Biology","confidence":"Medium","confidence_rationale":"Tier 2 — reciprocal co-IP plus ChIP; single lab but multiple mechanistic readouts","pmids":["33388549"],"is_preprint":false},{"year":2021,"finding":"Cryo-EM structure of human WLS in complex with WNT3A at 2.2 Å reveals that the WLS transmembrane domain adopts a GPCR-like fold with a conserved core cavity and lateral opening; the palmitoyleoyl lipid moiety on WNT3A (at its conserved palmitoleoylation site on a β-hairpin) threads through a hydrophobic tunnel of the WLS transmembrane domain and inserts into the membrane; this WNT3A β-hairpin–WLS interaction is essential for WLS-mediated Wnt secretion.","method":"Cryo-EM structure determination (2.2 Å resolution), functional validation of lipid-binding contacts, mutagenesis of WNT3A palmitoleoylation site","journal":"Nature Communications","confidence":"High","confidence_rationale":"Tier 1 — high-resolution cryo-EM structure with mutagenesis validation; defines atomic mechanism of WNT3A–WLS interaction","pmids":["34315898"],"is_preprint":false},{"year":2018,"finding":"WNT3A assembles into high-molecular-weight homo-trimeric and larger complexes in serum-containing media; binding to the extracellular CRD of Frizzled8 or to sFRP2 dissociates these complexes; assembly restricts WNT3A diffusion range in Xenopus embryos as shown by fluorescence correlation spectroscopy and immunohistochemistry.","method":"Analytical ultracentrifugation with fluorescence detection (AUC-FDS), gel filtration + cross-linking + single-particle analysis, fluorescence correlation spectroscopy (FCS), immunohistochemistry in Xenopus embryos","journal":"Communications Biology","confidence":"High","confidence_rationale":"Tier 1 — multiple biophysical methods (AUC, FCS, single-particle EM) combined with in vivo diffusion measurement; mechanistically rigorous","pmids":["30320232"],"is_preprint":false},{"year":2016,"finding":"Mechanical strain and WNT3A/β-catenin signaling synergize to drive cells through mitosis: strain alone induces Src-dependent Y654 phosphorylation of β-catenin and S/G2 accumulation without division; addition of WNT3A (or CKI inhibition) raises β-catenin-mediated transcription to a threshold required to trigger mitosis in strained cells.","method":"Mechanical strain device on MDCK cells, Casein Kinase I inhibitor, WNT3A addition, β-catenin Y654 phospho-specific antibody, cell cycle analysis, β-catenin reporter assay","journal":"eLife","confidence":"Medium","confidence_rationale":"Tier 2 — quantitative mechanistic dissection combining mechanical and biochemical perturbations; single lab","pmids":["27782880"],"is_preprint":false},{"year":2013,"finding":"PDGF receptor transactivation is the predominant β-catenin-independent mechanism by which WNT3A stimulates osteoblastic cell proliferation: WNT3A activates Src family kinases (SFKs) via Dishevelled, which transactivate PDGF receptors; soluble decoy PDGF-R excludes autocrine PDGF as the source; siRNA knockdown of PDGF-R confirms its predominant role.","method":"Selective kinase inhibitors (SFK, PDGF-R, PKC), DKK1 to exclude β-catenin contribution, PDGF-R siRNA, soluble decoy PDGF-R, time-course phosphorylation analysis in MC3T3-E1 cells","journal":"Journal of Bone and Mineral Research","confidence":"Medium","confidence_rationale":"Tier 2 — multiple pharmacological and genetic epistasis approaches; single lab but internally consistent","pmids":["22927028"],"is_preprint":false},{"year":2007,"finding":"WNT3A transcriptionally induces Lef-1 gene expression in airway submucosal gland progenitor cells in vivo through TCF4/β-catenin binding to a Wnt-responsive region in the Lef-1 promoter; ChIP confirmed TCF4 occupancy, and Wnt3a-deficient mice show loss of Lef-1 induction in forming gland buds.","method":"ChIP for TCF4 on Lef-1 promoter in primary airway epithelial cells, Lef-1 promoter–GFP reporter mice crossed to Wnt3a-deficient mice, dominant-active β-catenin + TCF4 transfection luciferase assay","journal":"Developmental Biology","confidence":"High","confidence_rationale":"Tier 2 — ChIP with in vivo genetic validation in Wnt3a KO mice; mechanistic target gene regulation established","pmids":["17335794"],"is_preprint":false},{"year":2010,"finding":"Sox17 directly represses WNT3A/β-catenin-mediated transcriptional activation of the Lef-1 promoter; EMSA and ChIP identified four Sox17-binding sites in the Lef-1 promoter that collaborate with TCF4 sites; Sox17–TCF complexes form through protein–protein interaction via Sox17's DNA- or β-catenin-binding domains.","method":"EMSA, ChIP, site-directed mutagenesis of Sox17 and TCF binding sites, luciferase reporter assay in primary airway epithelial cells and cell lines","journal":"American Journal of Physiology – Lung Cellular and Molecular Physiology","confidence":"Medium","confidence_rationale":"Tier 2 — ChIP plus EMSA plus combinatorial site mutagenesis; single lab","pmids":["20802155"],"is_preprint":false},{"year":2017,"finding":"WNT3A induces AChE expression in osteoblasts through a β-catenin–Runx2 axis; Wnt3a treatment increases Runx2 protein which binds a defined Runx2-binding site in the ACHE promoter (confirmed by ChIP), and deletion of this site reduces promoter activity; DKK-1 blocks the WNT3A-induced AChE expression.","method":"ChIP for Runx2 on ACHE promoter, ACHE promoter luciferase reporter with deletion mutants, Runx2 overexpression, DKK-1 inhibition, primary rat osteoblast culture","journal":"Journal of Biological Chemistry","confidence":"Medium","confidence_rationale":"Tier 2 — ChIP plus promoter mutagenesis and functional rescue; single lab","pmids":["28607150"],"is_preprint":false},{"year":2017,"finding":"LRP6, not LRP5, is the primary co-receptor mediating WNT3A canonical signaling in osteoblasts; genome-wide RNA-seq in Lrp5- and Lrp6-deficient primary calvarial osteoblasts treated with WNT3A identified 782 WNT3A-regulated genes, with LRP6 loss abolishing the majority of WNT3A transcriptional responses while LRP5 loss had minor effects.","method":"RNA-seq in Lrp5-KO and Lrp6-KO primary osteoblasts treated with WNT3A, receptor-specific genetic deletion","journal":"PLOS ONE","confidence":"Medium","confidence_rationale":"Tier 2 — genome-wide transcriptomics with co-receptor-specific knockouts; single lab","pmids":["29176883"],"is_preprint":false},{"year":2017,"finding":"WNT3A non-canonically triggers Ca2+ influx and CaMKII activation in differentiating human neural progenitor cells; elevated intracellular Ca2+ activates Pyk2, which in turn phosphorylates GSK-3β and stabilizes β-catenin, revealing a Wnt/Ca2+→Pyk2→GSK-3β→β-catenin cross-talk pathway.","method":"Intracellular Ca2+ imaging, CaMKII activity assay, Pyk2 phosphorylation Western blot, GSK-3β and β-catenin phosphorylation analysis in differentiating hNPCs","journal":"Biochemical and Biophysical Research Communications","confidence":"Medium","confidence_rationale":"Tier 2 — multiple sequential pathway components measured; single lab","pmids":["28694190"],"is_preprint":false},{"year":2019,"finding":"TLR4 activation increases Wnt3a and Dvl3 expression in monocytes; Wnt3a–Dvl3 signaling restrains TLR4-induced inflammatory responses by stabilizing β-catenin and suppressing NF-κB activity; siRNA or ectopic expression of Dvl3, GSK3β, and β-catenin confirmed the pathway; in vivo, Wnt3a inhibition during endotoxemia enhances systemic cytokines and neutrophil infiltration.","method":"siRNA knockdown and ectopic expression of Wnt3a, Dvl3, GSK3β, β-catenin in primary monocytes; ELISA, Western blot, NF-κB DNA binding assay; murine endotoxemia model with Wnt3a inhibition","journal":"Molecular Immunology","confidence":"Medium","confidence_rationale":"Tier 2 — gain- and loss-of-function in primary cells plus in vivo model; single lab","pmids":["31884387"],"is_preprint":false},{"year":2020,"finding":"JAK3 phospho-inactivates the E3 ubiquitin ligase Nedd4-2, reducing ubiquitination of Wnt3 protein and thereby increasing Wnt3a protein levels; elevated Wnt3a then signals through Dvl3 and β-catenin to suppress NF-κB and pro-inflammatory cytokines in innate immune cells; JAK3 or Wnt3a inhibition in vivo worsens P. gingivalis-induced periodontal disease.","method":"Gain- and loss-of-function of JAK3, Nedd4-2 ubiquitination assay, Wnt3 co-immunoprecipitation with Nedd4-2, Dvl3 and β-catenin downstream analysis, in vivo periodontal disease mouse model","journal":"FASEB Journal","confidence":"Medium","confidence_rationale":"Tier 2 — ubiquitination assay plus co-IP plus in vivo model; single lab but multiple mechanistic steps defined","pmids":["32433819"],"is_preprint":false},{"year":2015,"finding":"WNT3A activates MyoD expression by directly targeting cis-regulatory elements: WNT3A highly activates a composite proximal promoter + distal enhancer reporter and targets an upstream −8 to −9 kb Wnt-response element (L fragment) through Pax3/Pax7-binding sites; β-catenin and Pax7 binding to their respective sites in the distal enhancer and L fragment was confirmed by ChIP.","method":"Luciferase reporter deletion analysis of MyoD cis-elements, ChIP for β-catenin and Pax7, dominant-negative pathway constructs in HEK293T cells","journal":"Bioscience Reports","confidence":"Medium","confidence_rationale":"Tier 2 — ChIP plus systematic promoter/enhancer deletion mapping; single lab","pmids":["25651906"],"is_preprint":false},{"year":2014,"finding":"Intranasal WNT3A activates the Frizzled-1/PIWIL1a/FOXM1 pathway to reduce neuronal apoptosis after ischemic stroke; Frizzled-1 siRNA and PIWI1a siRNA reverse WNT3A neuroprotection, while PIWI1a CRISPR restoration after Frizzled-1 knockdown rescues FOXM1 and reduces cleaved caspase-3, placing Frizzled-1 upstream of PIWIL1a and FOXM1.","method":"Intranasal WNT3A administration in MCAO rats, Frizzled-1 siRNA, PIWI1a siRNA, PIWI1a CRISPR restoration, Western blot, immunohistochemistry, neurobehavioral testing, infarct volume measurement","journal":"Journal of Neuroscience","confidence":"Medium","confidence_rationale":"Tier 2 — genetic epistasis (siRNA + CRISPR rescue) with defined pathway order; single lab","pmids":["29954850"],"is_preprint":false},{"year":2014,"finding":"Rmb WNT3A normalizes altered hepatic lipogenesis and cholesterol biosynthesis in LRP6(R611C) mutant mice by reversing the Sp1-dependent IGF1/AKT/mTORC1/mTORC2 signaling activation that causes combined hyperlipidemia; in vitro rmWNT3A treatment of LRP6-mutant hepatocytes also restores pathway signaling to normal.","method":"In vivo rmWNT3A administration to LRP6(R611C) mice, in vitro primary hepatocyte treatment, IGF1R antagonist and rapamycin comparisons, plasma lipid and gene expression analysis","journal":"Cell Metabolism","confidence":"Medium","confidence_rationale":"Tier 2 — in vivo and in vitro rescue with rmWNT3A; pathway normalization validated by multiple enzyme expression readouts; single lab","pmids":["24506864"],"is_preprint":false},{"year":2021,"finding":"Localized WNT3A signaling induces asymmetric division of mouse embryonic stem cells; H3K27me3 (but not H3K4me3) modifications are correlated with gene expression changes that determine daughter cell fate after WNT3A-induced asymmetric division, as revealed by joint single-cell epigenome+transcriptome profiling.","method":"Same-cell epigenome and transcriptome sequencing (H3K27me3/H3K4me3 + gene expression), localized WNT3A stimulation of mouse ESCs","journal":"Nature Communications","confidence":"Medium","confidence_rationale":"Tier 2 — novel single-cell multiomic method applied to mechanistic question; single lab","pmids":["34642323"],"is_preprint":false},{"year":2020,"finding":"WNT3A-loaded exosomes activate canonical WNT signaling in intact cartilage in vivo (where soluble recombinant WNT3A fails), and a single injection improves osteochondral defect repair in mice over 8 weeks, demonstrating that exosomal delivery overcomes the hydrophobic barrier to WNT3A tissue penetration.","method":"TOPFlash reporter assay for in vitro WNT activity, TCF/Lef:H2B-GFP reporter mice for in vivo signaling, osteochondral defect mouse model, histological scoring","journal":"Journal of Extracellular Vesicles","confidence":"Medium","confidence_rationale":"Tier 2 — in vitro and in vivo comparison of soluble vs. exosomal WNT3A with pathway reporter; single lab","pmids":["34025953"],"is_preprint":false},{"year":2014,"finding":"WNT3A protein requires association with a lipid vesicle for in vivo stability and activity; liposome-reconstituted WNT3A (L-WNT3A) associates with the external lipid membrane surface, prolonging pathway activation in primary cells, and activates mesenchymal/skeletal stem cell populations in bone marrow engraftment assays in vivo.","method":"Liposome reconstitution of WNT3A, primary cell Wnt reporter assays, bone marrow engraftment in vivo assay, stem cell population analysis","journal":"PLOS ONE","confidence":"Medium","confidence_rationale":"Tier 2 — biophysical protein-lipid association combined with in vivo functional validation; single lab","pmids":["24400074"],"is_preprint":false},{"year":2020,"finding":"Nintedanib inhibits WNT3A-induced myofibroblast activation by blocking Src kinase activation and Src-dependent Y654 phosphorylation of β-catenin, thereby preventing β-catenin nuclear translocation; Src knockdown phenocopies nintedanib treatment, placing Src downstream of WNT3A and upstream of β-catenin nuclear entry in pulmonary fibroblasts.","method":"Nintedanib treatment, Src-specific siRNA knockdown, Src inhibitor KX2-391, β-catenin Y654 phosphorylation Western blot, nuclear fractionation, downstream Wnt target gene expression in lung fibroblasts","journal":"Frontiers in Pharmacology","confidence":"Medium","confidence_rationale":"Tier 2 — pharmacological and genetic (siRNA) epistasis; single lab but multiple orthogonal approaches","pmids":["32231574"],"is_preprint":false},{"year":2011,"finding":"Heparanase modulates WNT3A signaling in medulloblastoma cells by altering syndecan co-receptor function and GEF-H1 (a Rho-GEF) distribution; HPSE pretreatment differentially regulates Rac1/RhoA activities after WNT3A exposure, and the HPSE/HSPG axis modulates WNT3A-dependent β-catenin and N-Myc distribution as well as Gli transcription factor gene expression.","method":"Exogenous active heparanase treatment, Rac1/RhoA activity assays, immunofluorescence for GEF-H1/β-catenin, RT-PCR for Wnt target and HSPG genes, MB cell proliferation and invasion assays","journal":"Experimental and Therapeutic Medicine","confidence":"Low","confidence_rationale":"Tier 3 — single lab, correlative mechanistic data without direct binding/reconstitution of WNT3A–HSPG interaction","pmids":["21442027"],"is_preprint":false}],"current_model":"WNT3A is a palmitoyleoylated (Ser209), N-glycosylated secreted ligand that binds WLS via its lipid moiety within a hydrophobic transmembrane tunnel (cryo-EM structure resolved at 2.2 Å); vacuolar acidification releases WNT3A from the WLS complex for basolateral, clathrin/AP-1-dependent secretion, after which WNT3A assembles into diffusion-restricted homo-trimeric complexes that are dissociated by Frizzled-CRD or sFRP binding; upon receptor engagement, WNT3A acts as a bona fide GPCR ligand for Frizzled/Go/i proteins and signals through canonical β-catenin stabilization (requiring LEF-1/TCF nuclear effectors, and facilitated by ILK, RhoA/ROCK-mediated GSK-3β phosphorylation, and Pyk2-mediated Ca2+-dependent cross-talk), as well as β-catenin-independent activation of Raf-MEK-ERK, PI3K-Akt, RhoA/ROCK, and phosphoinositide (PI4K/PIP5K) pathways, collectively regulating paraxial mesoderm formation, left-right axis determination, segmentation, neural progenitor proliferation, osteoblast differentiation, and immune modulation."},"narrative":{"teleology":[{"year":1998,"claim":"Establishing that WNT3A functions as a diffusible paracrine signal: conditioned medium transfer showed that secreted WNT3A stabilizes β-catenin and remodels the actin cytoskeleton in recipient cells, providing the first demonstration that WNT3A has both canonical and cytoskeletal outputs.","evidence":"Conditioned medium from WNT3A-transfected L cells applied to C57MG cells; β-catenin Western blot and actin/adhesion protein immunostaining","pmids":["9893023"],"confidence":"High","gaps":["Receptor identity for WNT3A not yet determined","Mechanism of cytoskeletal reorganization downstream of WNT3A undefined","Whether WNT3A acts at long range in vivo unknown"]},{"year":1999,"claim":"Identifying the nuclear effectors: genetic epistasis showed that LEF-1 and TCF-1 double knockout phenocopies Wnt3a deficiency in mice, establishing them as essential downstream transcription factors for WNT3A in paraxial mesoderm and neural tube patterning.","evidence":"Lef1−/−;Tcf1−/− double-knockout mouse phenotypic comparison with Wnt3a−/− mice","pmids":["10090727"],"confidence":"High","gaps":["Whether additional TCF family members contribute in other tissues unknown","Signal transduction steps between WNT3A and LEF-1/TCF nuclear activation undefined"]},{"year":2004,"claim":"Resolving pathway bifurcation: WNT3A was shown to activate cell proliferation through two parallel arms — canonical β-catenin/TCF-4 and a β-catenin-independent Raf-1–MEK–ERK cascade — demonstrating that WNT3A is not solely a canonical Wnt ligand.","evidence":"β-catenin siRNA, MEK inhibitor U0126, dominant-negative TCF-4 in NIH3T3 cells with proliferation readouts","pmids":["15615777"],"confidence":"High","gaps":["Proximal receptor-level mechanism activating ERK independently of β-catenin not identified","Whether the two arms converge on shared gene targets unclear"]},{"year":2005,"claim":"Linking WNT3A to developmental patterning beyond mesoderm: Wnt3a knockout mice revealed that WNT3A from the primitive streak/dorsal node activates Delta/Notch to regulate perinodal Nodal expression for left-right axis determination while simultaneously controlling the segmentation clock.","evidence":"Wnt3a knockout mouse with in situ hybridization and genetic epistasis with Notch/Delta components","pmids":["16291790"],"confidence":"High","gaps":["Direct versus indirect activation of Delta/Notch by WNT3A not resolved","Whether WNT3A protein acts at long range to reach perinodal cells or requires relay unknown"]},{"year":2006,"claim":"Expanding the non-canonical signaling repertoire: WNT3A was found to activate PI3K–Akt independently of β-catenin and to require ILK for GSK-3β Ser9 phosphorylation and β-catenin stabilization, adding two new signaling branches to the WNT3A network.","evidence":"PI3K inhibitors and Akt siRNA in NIH3T3 cells; ILK pharmacological inhibition plus co-IP of ILK with APC/GSK-3β","pmids":["17011750","16799642"],"confidence":"High","gaps":["How ILK is activated by WNT3A receptor complex unknown","Whether PI3K–Akt and ILK pathways cross-talk in WNT3A context untested"]},{"year":2009,"claim":"Defining phosphoinositide and RhoA co-requirement: WNT3A was shown to activate PI4KIIα through Dvl to produce PtdIns(4,5)P₂ via a ternary Dvl–PI4KIIα–PIP5KI complex, and separately RhoA GTPase activity was found to be required for a subset of β-catenin-dependent WNT3A target genes and osteoblast differentiation.","evidence":"In vitro kinase reconstitution with purified PI4KIIα and Dvl; RhoA inhibition (C3 toxin) with microarray in C3H10T1/2 cells","pmids":["19561074","19482078"],"confidence":"High","gaps":["How Dvl is selectively directed to PI4KIIα versus other effectors unknown","Whether RhoA-dependent gene subset is tissue-specific untested"]},{"year":2010,"claim":"Elucidating the secretory mechanism: PORCN-dependent palmitoyleoylation at Ser209 was shown to be required for WNT3A–WLS binding, and vacuolar acidification was found necessary to release WNT3A from WLS into secretory vesicles, establishing the ER-to-vesicle transit mechanism.","evidence":"Ser209 mutagenesis, V-ATPase inhibitor treatment, WNT3A–WLS co-immunoprecipitation, cell fractionation","pmids":["20826466"],"confidence":"High","gaps":["Structural basis of pH-dependent WNT3A–WLS dissociation unknown at this stage","Post-WLS carriers of WNT3A not identified"]},{"year":2011,"claim":"Establishing Frizzled as a bona fide GPCR for WNT3A: direct GTPγS-binding assays demonstrated that WNT3A triggers Frizzled-mediated guanine-nucleotide exchange on Go/Gi proteins, blocked by pertussis toxin, proving GPCR-like signaling.","evidence":"GTPγS-binding assay on rat brain membranes and cultured cells with pertussis toxin and Wnt antagonist","pmids":["21128903"],"confidence":"High","gaps":["Which specific Frizzled family member(s) couple to Go/Gi upon WNT3A not resolved","Downstream effectors of Go/Gi in this context not identified"]},{"year":2013,"claim":"Defining polarized secretion and receptor transactivation: WNT3A was shown to undergo basolateral secretion via clathrin/AP-1 with two high-mannose N-glycans, and separately WNT3A was found to stimulate osteoblast proliferation through Src/Dishevelled-mediated PDGFR transactivation independent of β-catenin.","evidence":"Mass spectrometry glycan analysis with AP-1/AP-2 knockdown in MDCK cells; PDGFR siRNA and Src inhibitors in MC3T3-E1 osteoblasts","pmids":["23613470","22927028"],"confidence":"High","gaps":["Whether basolateral polarity of WNT3A secretion is universal across epithelia unknown","PDGFR transactivation mechanism (direct phosphorylation vs. adaptor) not molecularly defined"]},{"year":2016,"claim":"Reconstituting the RhoA/ROCK–GSK-3β axis and mechanical synergy: in vitro kinase assays showed ROCK directly phosphorylates GSK-3β Ser9, establishing a RhoA/ROCK branch that feeds into canonical β-catenin stabilization; concurrently, mechanical strain was shown to synergize with WNT3A to drive mitosis via a β-catenin transcriptional threshold mechanism.","evidence":"Purified ROCK kinase assay on recombinant GSK-3β with shRNA validation; mechanical strain device on MDCK cells with WNT3A addition and cell cycle analysis","pmids":["27575935","27782880"],"confidence":"High","gaps":["Whether ROCK-mediated GSK-3β phosphorylation operates in all WNT3A-responsive tissues untested","How mechanical strain integrates with WNT3A at the receptor level unknown"]},{"year":2017,"claim":"Clarifying co-receptor specificity and Ca²⁺ cross-talk: genome-wide RNA-seq in Lrp5/Lrp6-knockout osteoblasts revealed LRP6 as the primary canonical co-receptor for WNT3A, and in neural progenitors WNT3A was found to trigger Ca²⁺ influx activating Pyk2, which phosphorylates GSK-3β to stabilize β-catenin, bridging non-canonical Ca²⁺ and canonical pathways.","evidence":"RNA-seq in Lrp5-KO and Lrp6-KO primary osteoblasts; Ca²⁺ imaging and Pyk2 phosphorylation analysis in human neural progenitor cells","pmids":["29176883","28694190"],"confidence":"Medium","gaps":["Whether LRP6 preference over LRP5 extends beyond osteoblasts unknown","Ca²⁺ channel identity activated by WNT3A not identified","Whether Pyk2-mediated cross-talk operates in non-neural cell types untested"]},{"year":2018,"claim":"Revealing oligomeric state and ROS-dependent nuclear RhoA–β-catenin complex: biophysical analyses demonstrated WNT3A forms homo-trimeric complexes that restrict diffusion and are dissociated by Frizzled-CRD or sFRP binding; separately, WNT3A-induced superoxide was shown to drive Src-mediated Tyr42 phosphorylation of RhoA, enabling RhoA–β-catenin nuclear translocation to the Vimentin promoter.","evidence":"Analytical ultracentrifugation, fluorescence correlation spectroscopy in Xenopus embryos; co-IP of p-Tyr42 RhoA with β-catenin and ChIP on Vim promoter","pmids":["30320232","33388549"],"confidence":"High","gaps":["Stoichiometry and lipid requirements for WNT3A trimerization not fully defined","Whether ROS-dependent RhoA–β-catenin complex is a general WNT3A mechanism or context-specific unknown"]},{"year":2019,"claim":"Defining WNT3A's anti-inflammatory role: WNT3A/Dvl3/β-catenin signaling was found to restrain TLR4-induced NF-κB activation in monocytes, with JAK3-mediated stabilization of Wnt3 protein (via Nedd4-2 inactivation) amplifying this pathway; in vivo Wnt3a inhibition exacerbated endotoxemia and periodontal disease.","evidence":"siRNA/ectopic expression of Wnt3a, Dvl3, β-catenin in primary monocytes; JAK3–Nedd4-2 ubiquitination assays; murine endotoxemia and periodontal disease models","pmids":["31884387","32433819"],"confidence":"Medium","gaps":["Whether WNT3A anti-inflammatory function is Frizzled subtype-specific untested","Direct binding of Nedd4-2 to WNT3A protein not demonstrated"]},{"year":2021,"claim":"Atomic resolution of the WNT3A–WLS complex and epigenetic outputs: cryo-EM at 2.2 Å revealed the palmitoyleoyl lipid threading through a GPCR-like WLS transmembrane tunnel, establishing the structural basis of WNT secretion; concurrently, single-cell multi-omic profiling showed WNT3A-driven asymmetric stem cell division correlates with H3K27me3 redistribution.","evidence":"Cryo-EM structure with mutagenesis validation; same-cell epigenome+transcriptome sequencing in mouse ESCs","pmids":["34315898","34642323"],"confidence":"High","gaps":["Structural basis of pH-dependent WNT3A release from WLS not captured in current structure","Whether H3K27me3 changes are directly instructed by β-catenin/TCF or secondary unknown","Structure of WNT3A bound to Frizzled CRD together with LRP6 not available"]},{"year":null,"claim":"Major unresolved questions include: the structural basis of WNT3A engagement with the Frizzled–LRP6 holoreceptor complex, the mechanism by which Frizzled–Go/Gi coupling is selected over β-catenin-dependent versus -independent outputs, how WNT3A homo-trimers are formed and regulated in vivo, and whether the multiple β-catenin-independent branches (ERK, PI3K-Akt, RhoA, PDGFR transactivation, Ca²⁺/Pyk2) operate simultaneously or are context-selectively activated.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No ternary WNT3A–Frizzled–LRP6 structure exists","Selectivity mechanism for canonical versus non-canonical pathway activation undefined","In vivo regulation of WNT3A oligomeric state not established"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0048018","term_label":"receptor ligand activity","supporting_discovery_ids":[1,8,14]},{"term_id":"GO:0060089","term_label":"molecular transducer activity","supporting_discovery_ids":[8]},{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[2,7,11,22]}],"localization":[{"term_id":"GO:0005576","term_label":"extracellular region","supporting_discovery_ids":[1,14,28,29]},{"term_id":"GO:0005783","term_label":"endoplasmic reticulum","supporting_discovery_ids":[3,13]},{"term_id":"GO:0031410","term_label":"cytoplasmic vesicle","supporting_discovery_ids":[9,28]}],"pathway":[{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[0,1,2,5,6,8,10,11,21]},{"term_id":"R-HSA-1266738","term_label":"Developmental Biology","supporting_discovery_ids":[0,4,24,27]},{"term_id":"R-HSA-168256","term_label":"Immune System","supporting_discovery_ids":[22,23]},{"term_id":"R-HSA-9609507","term_label":"Protein localization","supporting_discovery_ids":[3,9,13]}],"complexes":["WNT3A–WLS secretory complex","WNT3A homo-trimer"],"partners":["WLS","LRP6","FZD1","FZD8","DVL3","ILK","SFRP2"],"other_free_text":[]},"mechanistic_narrative":"WNT3A is a palmitoyleoylated, N-glycosylated secreted ligand that activates both canonical β-catenin and multiple β-catenin-independent signaling cascades to regulate paraxial mesoderm formation, left-right axis determination, somite segmentation, osteoblast differentiation, neural progenitor proliferation, and innate immune modulation [PMID:10090727, PMID:16291790, PMID:19482078, PMID:31884387]. The lipid moiety at Ser209, added by PORCN, threads through a hydrophobic tunnel in the WLS transmembrane domain (resolved at 2.2 Å by cryo-EM) to mediate ER-to-plasma-membrane trafficking via clathrin/AP-1-dependent basolateral secretion; extracellularly, WNT3A assembles into diffusion-restricted homo-trimeric complexes that are dissociated by Frizzled-CRD or sFRP binding [PMID:20826466, PMID:34315898, PMID:23613470, PMID:30320232]. Upon receptor engagement, WNT3A functions as a GPCR ligand coupling Frizzled to Go/Gi proteins and signals through LRP6-dependent β-catenin stabilization (requiring LEF-1/TCF nuclear effectors and facilitated by ILK, RhoA/ROCK-mediated GSK-3β phosphorylation, and Pyk2-mediated Ca²⁺ cross-talk), as well as β-catenin-independent Raf-MEK-ERK, PI3K-Akt, Src-PDGFR transactivation, and PI4KIIα/PIP5KI phosphoinositide pathways [PMID:21128903, PMID:15615777, PMID:16799642, PMID:27575935, PMID:28694190, PMID:19561074, PMID:22927028, PMID:29176883]. WNT3A-driven β-catenin transcriptional activity, modulated by H3K27me3 epigenetic marks, also directs asymmetric stem cell division and cooperates with mechanical strain to trigger mitosis [PMID:34642323, PMID:27782880]."},"prefetch_data":{"uniprot":{"accession":"P56704","full_name":"Protein Wnt-3a","aliases":[],"length_aa":352,"mass_kda":39.4,"function":"Ligand for members of the frizzled family of seven transmembrane receptors (Probable). Functions in the canonical Wnt signaling pathway that results in activation of transcription factors of the TCF/LEF family (PubMed:20093360, PubMed:21244856, PubMed:24841207, PubMed:26902720). Required for normal embryonic mesoderm development and formation of caudal somites. Required for normal morphogenesis of the developing neural tube (By similarity). Mediates self-renewal of the stem cells at the bottom on intestinal crypts (in vitro) (PubMed:26902720)","subcellular_location":"Secreted, extracellular space, extracellular matrix; Secreted","url":"https://www.uniprot.org/uniprotkb/P56704/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/WNT3A","classification":"Not Classified","n_dependent_lines":1,"n_total_lines":1208,"dependency_fraction":0.0008278145695364238},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/WNT3A","total_profiled":1310},"omim":[{"mim_id":"621260","title":"SHORT-RIB THORACIC DYSPLASIA 22 WITHOUT POLYDACTYLY; SRTD22","url":"https://www.omim.org/entry/621260"},{"mim_id":"618942","title":"IQ MOTIF- AND ANKYRIN REPEAT-CONTAINING PROTEIN 1; IQANK1","url":"https://www.omim.org/entry/618942"},{"mim_id":"618649","title":"HECT DOMAIN E3 UBIQUITIN PROTEIN LIGASE 1; HECTD1","url":"https://www.omim.org/entry/618649"},{"mim_id":"617875","title":"POLYCYSTIC LIVER DISEASE 4 WITH OR WITHOUT KIDNEY CYSTS; PCLD4","url":"https://www.omim.org/entry/617875"},{"mim_id":"617595","title":"BIRK-LANDAU-PEREZ SYNDROME; BILAPES","url":"https://www.omim.org/entry/617595"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Approved","locations":[{"location":"Vesicles","reliability":"Approved"}],"tissue_specificity":"Tissue enriched","tissue_distribution":"Detected in some","driving_tissues":[{"tissue":"placenta","ntpm":14.7}],"url":"https://www.proteinatlas.org/search/WNT3A"},"hgnc":{"alias_symbol":[],"prev_symbol":[]},"alphafold":{"accession":"P56704","domains":[{"cath_id":"-","chopping":"19-135_159-188_223-291","consensus_level":"high","plddt":89.444,"start":19,"end":291},{"cath_id":"3.30.2460.20","chopping":"296-352","consensus_level":"medium","plddt":94.9696,"start":296,"end":352}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/P56704","model_url":"https://alphafold.ebi.ac.uk/files/AF-P56704-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-P56704-F1-predicted_aligned_error_v6.png","plddt_mean":88.31},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=WNT3A","jax_strain_url":"https://www.jax.org/strain/search?query=WNT3A"},"sequence":{"accession":"P56704","fasta_url":"https://rest.uniprot.org/uniprotkb/P56704.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/P56704/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/P56704"}},"corpus_meta":[{"pmid":"10090727","id":"PMC_10090727","title":"Wnt3a-/--like phenotype and limb deficiency in Lef1(-/-)Tcf1(-/-) mice.","date":"1999","source":"Genes & development","url":"https://pubmed.ncbi.nlm.nih.gov/10090727","citation_count":403,"is_preprint":false},{"pmid":"9893023","id":"PMC_9893023","title":"Cytoskeletal reorganization by soluble Wnt-3a protein signalling.","date":"1998","source":"Genes to cells : devoted to molecular & cellular mechanisms","url":"https://pubmed.ncbi.nlm.nih.gov/9893023","citation_count":233,"is_preprint":false},{"pmid":"15615777","id":"PMC_15615777","title":"Both ERK and Wnt/beta-catenin pathways are involved in Wnt3a-induced proliferation.","date":"2004","source":"Journal of cell science","url":"https://pubmed.ncbi.nlm.nih.gov/15615777","citation_count":171,"is_preprint":false},{"pmid":"20826466","id":"PMC_20826466","title":"WLS-dependent secretion of WNT3A requires Ser209 acylation and vacuolar acidification.","date":"2010","source":"Journal of cell science","url":"https://pubmed.ncbi.nlm.nih.gov/20826466","citation_count":162,"is_preprint":false},{"pmid":"16291790","id":"PMC_16291790","title":"Wnt3a links left-right determination with segmentation and anteroposterior axis elongation.","date":"2005","source":"Development (Cambridge, England)","url":"https://pubmed.ncbi.nlm.nih.gov/16291790","citation_count":137,"is_preprint":false},{"pmid":"25499541","id":"PMC_25499541","title":"Wnt3a expression is associated with epithelial-mesenchymal transition and promotes colon cancer progression.","date":"2014","source":"Journal of experimental & clinical cancer research : CR","url":"https://pubmed.ncbi.nlm.nih.gov/25499541","citation_count":108,"is_preprint":false},{"pmid":"29659176","id":"PMC_29659176","title":"Nrf2 attenuates inflammatory response in COPD/emphysema: Crosstalk with Wnt3a/β-catenin and AMPK pathways.","date":"2018","source":"Journal of cellular and molecular medicine","url":"https://pubmed.ncbi.nlm.nih.gov/29659176","citation_count":106,"is_preprint":false},{"pmid":"24506864","id":"PMC_24506864","title":"The combined hyperlipidemia caused by impaired Wnt-LRP6 signaling is reversed by Wnt3a rescue.","date":"2014","source":"Cell metabolism","url":"https://pubmed.ncbi.nlm.nih.gov/24506864","citation_count":95,"is_preprint":false},{"pmid":"24080158","id":"PMC_24080158","title":"Secreted Frizzled-related protein potentiation versus inhibition of Wnt3a/β-catenin signaling.","date":"2013","source":"Cellular signalling","url":"https://pubmed.ncbi.nlm.nih.gov/24080158","citation_count":83,"is_preprint":false},{"pmid":"16799642","id":"PMC_16799642","title":"Modulation of Wnt3a-mediated nuclear beta-catenin accumulation and activation by integrin-linked kinase in mammalian cells.","date":"2006","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/16799642","citation_count":83,"is_preprint":false},{"pmid":"26930594","id":"PMC_26930594","title":"TLR4 Activation Promotes Bone Marrow MSC Proliferation and Osteogenic Differentiation via Wnt3a and Wnt5a Signaling.","date":"2016","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/26930594","citation_count":80,"is_preprint":false},{"pmid":"30889487","id":"PMC_30889487","title":"Captopril attenuates TAC-induced heart failure via inhibiting Wnt3a/β-catenin and Jak2/Stat3 pathways.","date":"2019","source":"Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie","url":"https://pubmed.ncbi.nlm.nih.gov/30889487","citation_count":79,"is_preprint":false},{"pmid":"31178968","id":"PMC_31178968","title":"Overexpression of miR-27b-3p Targeting Wnt3a Regulates the Signaling Pathway of Wnt/β-Catenin and Attenuates Atrial Fibrosis in Rats with Atrial Fibrillation.","date":"2019","source":"Oxidative medicine and cellular longevity","url":"https://pubmed.ncbi.nlm.nih.gov/31178968","citation_count":78,"is_preprint":false},{"pmid":"15892713","id":"PMC_15892713","title":"WNT1 and WNT3a promote expansion of melanocytes through distinct modes of action.","date":"2005","source":"Pigment cell research","url":"https://pubmed.ncbi.nlm.nih.gov/15892713","citation_count":69,"is_preprint":false},{"pmid":"21128903","id":"PMC_21128903","title":"Wnt3a stimulation elicits G-protein-coupled receptor properties of mammalian Frizzled proteins.","date":"2011","source":"The Biochemical journal","url":"https://pubmed.ncbi.nlm.nih.gov/21128903","citation_count":69,"is_preprint":false},{"pmid":"29164582","id":"PMC_29164582","title":"miR-15a-5p suppresses endometrial cancer cell growth via Wnt/β-catenin signaling pathway by inhibiting WNT3A.","date":"2017","source":"European review for medical and pharmacological sciences","url":"https://pubmed.ncbi.nlm.nih.gov/29164582","citation_count":67,"is_preprint":false},{"pmid":"25848952","id":"PMC_25848952","title":"Transcriptome analysis of Wnt3a-treated triple-negative breast cancer cells.","date":"2015","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/25848952","citation_count":66,"is_preprint":false},{"pmid":"17011750","id":"PMC_17011750","title":"The PI3 kinase-Akt pathway mediates Wnt3a-induced proliferation.","date":"2006","source":"Cellular signalling","url":"https://pubmed.ncbi.nlm.nih.gov/17011750","citation_count":62,"is_preprint":false},{"pmid":"19561074","id":"PMC_19561074","title":"Regulation of phosphatidylinositol kinases and metabolism by Wnt3a and Dvl.","date":"2009","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/19561074","citation_count":59,"is_preprint":false},{"pmid":"23613470","id":"PMC_23613470","title":"The apical and basolateral secretion of Wnt11 and Wnt3a in polarized epithelial cells is regulated by different mechanisms.","date":"2013","source":"Journal of cell science","url":"https://pubmed.ncbi.nlm.nih.gov/23613470","citation_count":59,"is_preprint":false},{"pmid":"19523451","id":"PMC_19523451","title":"Wnt3a regulates proliferation and migration of HUVEC via canonical and non-canonical Wnt signaling pathways.","date":"2009","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/19523451","citation_count":58,"is_preprint":false},{"pmid":"29954850","id":"PMC_29954850","title":"Intranasal wnt3a Attenuates Neuronal Apoptosis through Frz1/PIWIL1a/FOXM1 Pathway in MCAO Rats.","date":"2018","source":"The Journal of neuroscience : the official journal of the Society for Neuroscience","url":"https://pubmed.ncbi.nlm.nih.gov/29954850","citation_count":52,"is_preprint":false},{"pmid":"29948330","id":"PMC_29948330","title":"MiR-195 suppresses colon cancer proliferation and metastasis by targeting WNT3A.","date":"2018","source":"Molecular genetics and genomics : MGG","url":"https://pubmed.ncbi.nlm.nih.gov/29948330","citation_count":52,"is_preprint":false},{"pmid":"24400074","id":"PMC_24400074","title":"Drugging a stem cell compartment using Wnt3a protein as a therapeutic.","date":"2014","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/24400074","citation_count":50,"is_preprint":false},{"pmid":"27575935","id":"PMC_27575935","title":"Wnt3A Induces GSK-3β Phosphorylation and β-Catenin Accumulation Through RhoA/ROCK.","date":"2016","source":"Journal of cellular physiology","url":"https://pubmed.ncbi.nlm.nih.gov/27575935","citation_count":48,"is_preprint":false},{"pmid":"29948952","id":"PMC_29948952","title":"GBM-Derived Wnt3a Induces M2-Like Phenotype in Microglial Cells Through Wnt/β-Catenin Signaling.","date":"2018","source":"Molecular neurobiology","url":"https://pubmed.ncbi.nlm.nih.gov/29948952","citation_count":46,"is_preprint":false},{"pmid":"22151902","id":"PMC_22151902","title":"Effects of Wnt3A and mechanical load on cartilage chondrocyte homeostasis.","date":"2011","source":"Arthritis research & therapy","url":"https://pubmed.ncbi.nlm.nih.gov/22151902","citation_count":46,"is_preprint":false},{"pmid":"34025953","id":"PMC_34025953","title":"WNT3A-loaded exosomes enable cartilage repair.","date":"2021","source":"Journal of extracellular vesicles","url":"https://pubmed.ncbi.nlm.nih.gov/34025953","citation_count":46,"is_preprint":false},{"pmid":"19482078","id":"PMC_19482078","title":"Rho GTPase activity modulates Wnt3a/beta-catenin signaling.","date":"2009","source":"Cellular signalling","url":"https://pubmed.ncbi.nlm.nih.gov/19482078","citation_count":45,"is_preprint":false},{"pmid":"17335794","id":"PMC_17335794","title":"Wnt3a regulates Lef-1 expression during airway submucosal gland morphogenesis.","date":"2007","source":"Developmental biology","url":"https://pubmed.ncbi.nlm.nih.gov/17335794","citation_count":45,"is_preprint":false},{"pmid":"19638421","id":"PMC_19638421","title":"Folate rescues lithium-, homocysteine- and Wnt3A-induced vertebrate cardiac anomalies.","date":"2009","source":"Disease models & mechanisms","url":"https://pubmed.ncbi.nlm.nih.gov/19638421","citation_count":44,"is_preprint":false},{"pmid":"12011973","id":"PMC_12011973","title":"WNT3-WNT14B and WNT3A-WNT14 gene clusters (Review).","date":"2002","source":"International journal of molecular medicine","url":"https://pubmed.ncbi.nlm.nih.gov/12011973","citation_count":42,"is_preprint":false},{"pmid":"37451151","id":"PMC_37451151","title":"Quercetin induces MGMT+ glioblastoma cells apoptosis via dual inhibition of Wnt3a/β-Catenin and Akt/NF-κB signaling pathways.","date":"2023","source":"Phytomedicine : international journal of phytotherapy and phytopharmacology","url":"https://pubmed.ncbi.nlm.nih.gov/37451151","citation_count":42,"is_preprint":false},{"pmid":"25682158","id":"PMC_25682158","title":"Reengineering autologous bone grafts with the stem cell activator WNT3A.","date":"2015","source":"Biomaterials","url":"https://pubmed.ncbi.nlm.nih.gov/25682158","citation_count":42,"is_preprint":false},{"pmid":"32371389","id":"PMC_32371389","title":"BCP crystals promote chondrocyte hypertrophic differentiation in OA cartilage by sequestering Wnt3a.","date":"2020","source":"Annals of the rheumatic diseases","url":"https://pubmed.ncbi.nlm.nih.gov/32371389","citation_count":41,"is_preprint":false},{"pmid":"24904070","id":"PMC_24904070","title":"Hippocampal Wnt3a is Necessary and Sufficient for Contextual Fear Memory Acquisition and Consolidation.","date":"2014","source":"Cerebral cortex (New York, N.Y. : 1991)","url":"https://pubmed.ncbi.nlm.nih.gov/24904070","citation_count":40,"is_preprint":false},{"pmid":"30145863","id":"PMC_30145863","title":"MicroRNA-766-3p Inhibits Tumour Progression by Targeting Wnt3a in Hepatocellular Carcinoma.","date":"2018","source":"Molecules and cells","url":"https://pubmed.ncbi.nlm.nih.gov/30145863","citation_count":40,"is_preprint":false},{"pmid":"34671013","id":"PMC_34671013","title":"GOLPH3/CKAP4 promotes metastasis and tumorigenicity by enhancing the secretion of exosomal WNT3A in non-small-cell lung cancer.","date":"2021","source":"Cell death & disease","url":"https://pubmed.ncbi.nlm.nih.gov/34671013","citation_count":40,"is_preprint":false},{"pmid":"27782880","id":"PMC_27782880","title":"Increasing β-catenin/Wnt3A activity levels drive mechanical strain-induced cell cycle progression through mitosis.","date":"2016","source":"eLife","url":"https://pubmed.ncbi.nlm.nih.gov/27782880","citation_count":39,"is_preprint":false},{"pmid":"31043858","id":"PMC_31043858","title":"PITX2 enhances progression of lung adenocarcinoma by transcriptionally regulating WNT3A and activating Wnt/β-catenin signaling pathway.","date":"2019","source":"Cancer cell international","url":"https://pubmed.ncbi.nlm.nih.gov/31043858","citation_count":37,"is_preprint":false},{"pmid":"24808138","id":"PMC_24808138","title":"Smad1 transcription factor integrates BMP2 and Wnt3a signals in migrating cardiac progenitor cells.","date":"2014","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/24808138","citation_count":36,"is_preprint":false},{"pmid":"31111621","id":"PMC_31111621","title":"Wnt3a promotes radioresistance via autophagy in squamous cell carcinoma of the head and neck.","date":"2019","source":"Journal of cellular and molecular medicine","url":"https://pubmed.ncbi.nlm.nih.gov/31111621","citation_count":35,"is_preprint":false},{"pmid":"29740905","id":"PMC_29740905","title":"Involvement of miR-140-3p in Wnt3a and TGFβ3 signaling pathways during osteoblast differentiation in MC3T3-E1 cells.","date":"2018","source":"Genes to cells : devoted to molecular & cellular mechanisms","url":"https://pubmed.ncbi.nlm.nih.gov/29740905","citation_count":35,"is_preprint":false},{"pmid":"20589426","id":"PMC_20589426","title":"Wnt3a promotes hippocampal neurogenesis by shortening cell cycle duration of neural progenitor cells.","date":"2010","source":"Cellular and molecular neurobiology","url":"https://pubmed.ncbi.nlm.nih.gov/20589426","citation_count":34,"is_preprint":false},{"pmid":"34642323","id":"PMC_34642323","title":"Joint single-cell multiomic analysis in Wnt3a induced asymmetric stem cell division.","date":"2021","source":"Nature communications","url":"https://pubmed.ncbi.nlm.nih.gov/34642323","citation_count":33,"is_preprint":false},{"pmid":"25809880","id":"PMC_25809880","title":"Wnt8a and Wnt3a cooperate in the axial stem cell niche to promote mammalian body axis extension.","date":"2015","source":"Developmental dynamics : an official publication of the American Association of Anatomists","url":"https://pubmed.ncbi.nlm.nih.gov/25809880","citation_count":32,"is_preprint":false},{"pmid":"29176883","id":"PMC_29176883","title":"Wnt co-receptors Lrp5 and Lrp6 differentially mediate Wnt3a signaling in osteoblasts.","date":"2017","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/29176883","citation_count":32,"is_preprint":false},{"pmid":"34315898","id":"PMC_34315898","title":"Cryo-EM structure of human Wntless in complex with Wnt3a.","date":"2021","source":"Nature communications","url":"https://pubmed.ncbi.nlm.nih.gov/34315898","citation_count":32,"is_preprint":false},{"pmid":"30720151","id":"PMC_30720151","title":"MicroRNA-9-5p promotes osteoporosis development through inhibiting osteogenesis and promoting adipogenesis via targeting Wnt3a.","date":"2019","source":"European review for medical and pharmacological sciences","url":"https://pubmed.ncbi.nlm.nih.gov/30720151","citation_count":32,"is_preprint":false},{"pmid":"25084050","id":"PMC_25084050","title":"WNT3A promotes myogenesis of human embryonic stem cells and enhances in vivo engraftment.","date":"2014","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/25084050","citation_count":31,"is_preprint":false},{"pmid":"33388549","id":"PMC_33388549","title":"RhoA GTPase phosphorylated at tyrosine 42 by src kinase binds to β-catenin and contributes transcriptional regulation of vimentin upon Wnt3A.","date":"2020","source":"Redox biology","url":"https://pubmed.ncbi.nlm.nih.gov/33388549","citation_count":30,"is_preprint":false},{"pmid":"28607150","id":"PMC_28607150","title":"Wnt3a induces the expression of acetylcholinesterase during osteoblast differentiation via the Runx2 transcription factor.","date":"2017","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/28607150","citation_count":29,"is_preprint":false},{"pmid":"25277232","id":"PMC_25277232","title":"R-spondin1 regulates cell proliferation of corneal endothelial cells via the Wnt3a/β-catenin pathway.","date":"2014","source":"Investigative ophthalmology & visual science","url":"https://pubmed.ncbi.nlm.nih.gov/25277232","citation_count":29,"is_preprint":false},{"pmid":"18242164","id":"PMC_18242164","title":"Effects of Wnt3a on proliferation and differentiation of human epidermal stem cells.","date":"2008","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/18242164","citation_count":29,"is_preprint":false},{"pmid":"26112214","id":"PMC_26112214","title":"Wnt5a attenuates Wnt3a-induced alkaline phosphatase expression in dental follicle cells.","date":"2015","source":"Experimental cell research","url":"https://pubmed.ncbi.nlm.nih.gov/26112214","citation_count":28,"is_preprint":false},{"pmid":"24535659","id":"PMC_24535659","title":"Wnt3a activates β1-integrin and regulates migration and adhesion of vascular smooth muscle cells.","date":"2014","source":"Molecular medicine reports","url":"https://pubmed.ncbi.nlm.nih.gov/24535659","citation_count":28,"is_preprint":false},{"pmid":"29588395","id":"PMC_29588395","title":"Substratum stiffness tunes proliferation downstream of Wnt3a in part by regulating integrin-linked kinase and frizzled-1.","date":"2018","source":"Journal of cell science","url":"https://pubmed.ncbi.nlm.nih.gov/29588395","citation_count":28,"is_preprint":false},{"pmid":"26006230","id":"PMC_26006230","title":"MicroRNA-27a-3p Inhibits Melanogenesis in Mouse Skin Melanocytes by Targeting Wnt3a.","date":"2015","source":"International journal of molecular sciences","url":"https://pubmed.ncbi.nlm.nih.gov/26006230","citation_count":28,"is_preprint":false},{"pmid":"31336380","id":"PMC_31336380","title":"Therapeutic Potential of Wnt-3a in Neurological Recovery after Spinal Cord Injury.","date":"2019","source":"European neurology","url":"https://pubmed.ncbi.nlm.nih.gov/31336380","citation_count":27,"is_preprint":false},{"pmid":"7660319","id":"PMC_7660319","title":"Interactions of Wnt-1 and Wnt-3a are essential for neural tube patterning.","date":"1995","source":"Teratology","url":"https://pubmed.ncbi.nlm.nih.gov/7660319","citation_count":27,"is_preprint":false},{"pmid":"24922070","id":"PMC_24922070","title":"Wnt3a mitigates acute lung injury by reducing P2X7 receptor-mediated alveolar epithelial type I cell death.","date":"2014","source":"Cell death & disease","url":"https://pubmed.ncbi.nlm.nih.gov/24922070","citation_count":26,"is_preprint":false},{"pmid":"20802155","id":"PMC_20802155","title":"Sox17 modulates Wnt3A/beta-catenin-mediated transcriptional activation of the Lef-1 promoter.","date":"2010","source":"American journal of physiology. Lung cellular and molecular physiology","url":"https://pubmed.ncbi.nlm.nih.gov/20802155","citation_count":26,"is_preprint":false},{"pmid":"30018042","id":"PMC_30018042","title":"Wnt3a Neutralization Enhances T-cell Responses through Indirect Mechanisms and Restrains Tumor Growth.","date":"2018","source":"Cancer immunology research","url":"https://pubmed.ncbi.nlm.nih.gov/30018042","citation_count":25,"is_preprint":false},{"pmid":"37072558","id":"PMC_37072558","title":"Wnt3a-Loaded Extracellular Vesicles Promote Alveolar Epithelial Regeneration after Lung Injury.","date":"2023","source":"Advanced science (Weinheim, Baden-Wurttemberg, Germany)","url":"https://pubmed.ncbi.nlm.nih.gov/37072558","citation_count":25,"is_preprint":false},{"pmid":"36657317","id":"PMC_36657317","title":"Achyranthoside D attenuates chondrocyte loss and inflammation in osteoarthritis via targeted regulation of Wnt3a.","date":"2023","source":"Phytomedicine : international journal of phytotherapy and phytopharmacology","url":"https://pubmed.ncbi.nlm.nih.gov/36657317","citation_count":25,"is_preprint":false},{"pmid":"21442027","id":"PMC_21442027","title":"Heparanase Modulates Shh and Wnt3a Signaling in Human Medulloblastoma Cells.","date":"2011","source":"Experimental and therapeutic medicine","url":"https://pubmed.ncbi.nlm.nih.gov/21442027","citation_count":24,"is_preprint":false},{"pmid":"30320232","id":"PMC_30320232","title":"Assembly of protein complexes restricts diffusion of Wnt3a proteins.","date":"2018","source":"Communications biology","url":"https://pubmed.ncbi.nlm.nih.gov/30320232","citation_count":24,"is_preprint":false},{"pmid":"31884387","id":"PMC_31884387","title":"TLR4 induced Wnt3a-Dvl3 restrains the intensity of inflammation and protects against endotoxin-driven organ failure through GSK3β/β-catenin signaling.","date":"2019","source":"Molecular immunology","url":"https://pubmed.ncbi.nlm.nih.gov/31884387","citation_count":24,"is_preprint":false},{"pmid":"27158593","id":"PMC_27158593","title":"Arsenic inhibits stem cell differentiation by altering the interplay between the Wnt3a and Notch signaling pathways.","date":"2016","source":"Toxicology reports","url":"https://pubmed.ncbi.nlm.nih.gov/27158593","citation_count":23,"is_preprint":false},{"pmid":"26414930","id":"PMC_26414930","title":"Expression of Wnt3a, Wnt10b, β-catenin and DKK1 in periodontium during orthodontic tooth movement in rats.","date":"2015","source":"Acta odontologica Scandinavica","url":"https://pubmed.ncbi.nlm.nih.gov/26414930","citation_count":23,"is_preprint":false},{"pmid":"30896984","id":"PMC_30896984","title":"Wnt3a Activates the WNT-YAP/TAZ Pathway to Sustain CDX2 Expression in Bovine Trophoblast Stem Cells.","date":"2019","source":"DNA and cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/30896984","citation_count":23,"is_preprint":false},{"pmid":"34502380","id":"PMC_34502380","title":"Piezoelectric Microvibration Mitigates Estrogen Loss-Induced Osteoporosis and Promotes Piezo1, MicroRNA-29a, and Wnt3a Signaling in Osteoblasts.","date":"2021","source":"International journal of molecular sciences","url":"https://pubmed.ncbi.nlm.nih.gov/34502380","citation_count":23,"is_preprint":false},{"pmid":"33128175","id":"PMC_33128175","title":"Blockers of Wnt3a, Wnt10a, or β-Catenin Prevent Chemotherapy-Induced Neuropathic Pain In Vivo.","date":"2020","source":"Neurotherapeutics : the journal of the American Society for Experimental NeuroTherapeutics","url":"https://pubmed.ncbi.nlm.nih.gov/33128175","citation_count":22,"is_preprint":false},{"pmid":"33684507","id":"PMC_33684507","title":"LGR5/R-Spo1/Wnt3a axis promotes stemness and aggressive phenotype in hepatoblast-like hepatocellular carcinoma cell lines.","date":"2021","source":"Cellular signalling","url":"https://pubmed.ncbi.nlm.nih.gov/33684507","citation_count":22,"is_preprint":false},{"pmid":"31412883","id":"PMC_31412883","title":"Apigenin inhibits fibroblast proliferation and reduces epidural fibrosis by regulating Wnt3a/β-catenin signaling pathway.","date":"2019","source":"Journal of orthopaedic surgery and research","url":"https://pubmed.ncbi.nlm.nih.gov/31412883","citation_count":22,"is_preprint":false},{"pmid":"26846714","id":"PMC_26846714","title":"Wnt3a suppresses Pseudomonas aeruginosa-induced inflammation and promotes bacterial killing in macrophages.","date":"2016","source":"Molecular medicine reports","url":"https://pubmed.ncbi.nlm.nih.gov/26846714","citation_count":22,"is_preprint":false},{"pmid":"31737122","id":"PMC_31737122","title":"Oncogenic Wnt3a: A Candidate Specific Marker and Novel Molecular Target for Hepatocellular Carcinoma.","date":"2019","source":"Journal of Cancer","url":"https://pubmed.ncbi.nlm.nih.gov/31737122","citation_count":22,"is_preprint":false},{"pmid":"32865704","id":"PMC_32865704","title":"CircDHDDS/miR-361-3p/WNT3A Axis Promotes the Development of Retinoblastoma by Regulating Proliferation, Cell Cycle, Migration, and Invasion of Retinoblastoma Cells.","date":"2020","source":"Neurochemical research","url":"https://pubmed.ncbi.nlm.nih.gov/32865704","citation_count":22,"is_preprint":false},{"pmid":"31613171","id":"PMC_31613171","title":"MiR-27a regulates WNT3A and KITLG expression in Cashmere goats with different coat colors.","date":"2019","source":"Animal biotechnology","url":"https://pubmed.ncbi.nlm.nih.gov/31613171","citation_count":21,"is_preprint":false},{"pmid":"22927028","id":"PMC_22927028","title":"Predominant role of PDGF receptor transactivation in Wnt3a-induced osteoblastic cell proliferation.","date":"2013","source":"Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research","url":"https://pubmed.ncbi.nlm.nih.gov/22927028","citation_count":21,"is_preprint":false},{"pmid":"37563362","id":"PMC_37563362","title":"LINC00665 activating Wnt3a/β-catenin signaling by bond with YBX1 promotes gastric cancer proliferation and metastasis.","date":"2023","source":"Cancer gene therapy","url":"https://pubmed.ncbi.nlm.nih.gov/37563362","citation_count":21,"is_preprint":false},{"pmid":"22399680","id":"PMC_22399680","title":"A hindbrain-repressive Wnt3a/Meis3/Tsh1 circuit promotes neuronal differentiation and coordinates tissue maturation.","date":"2012","source":"Development (Cambridge, England)","url":"https://pubmed.ncbi.nlm.nih.gov/22399680","citation_count":21,"is_preprint":false},{"pmid":"32231574","id":"PMC_32231574","title":"Nintedanib Inhibits Wnt3a-Induced Myofibroblast Activation by Suppressing the Src/β-Catenin Pathway.","date":"2020","source":"Frontiers in pharmacology","url":"https://pubmed.ncbi.nlm.nih.gov/32231574","citation_count":21,"is_preprint":false},{"pmid":"28463594","id":"PMC_28463594","title":"Effects of WNT3A and WNT16 on the Osteogenic and Adipogenic Differentiation of Perivascular Stem/Stromal Cells.","date":"2017","source":"Tissue engineering. Part A","url":"https://pubmed.ncbi.nlm.nih.gov/28463594","citation_count":20,"is_preprint":false},{"pmid":"32433819","id":"PMC_32433819","title":"JAK3 restrains inflammatory responses and protects against periodontal disease through Wnt3a signaling.","date":"2020","source":"FASEB journal : official publication of the Federation of American Societies for Experimental Biology","url":"https://pubmed.ncbi.nlm.nih.gov/32433819","citation_count":19,"is_preprint":false},{"pmid":"36633501","id":"PMC_36633501","title":"Wnt3a promotes odonto/osteogenic differentiation in vitro and tertiary dentin formation in a rat model.","date":"2023","source":"International endodontic journal","url":"https://pubmed.ncbi.nlm.nih.gov/36633501","citation_count":19,"is_preprint":false},{"pmid":"37307963","id":"PMC_37307963","title":"S1PR2/Wnt3a/RhoA/ROCK1/β-catenin signaling pathway promotes diabetic nephropathy by inducting endothelial mesenchymal transition and impairing endothelial barrier function.","date":"2023","source":"Life sciences","url":"https://pubmed.ncbi.nlm.nih.gov/37307963","citation_count":19,"is_preprint":false},{"pmid":"31236761","id":"PMC_31236761","title":"Role of Wnt3a in the pathogenesis of cancer, current status and prospective.","date":"2019","source":"Molecular biology reports","url":"https://pubmed.ncbi.nlm.nih.gov/31236761","citation_count":18,"is_preprint":false},{"pmid":"32570120","id":"PMC_32570120","title":"BAIAP2L2 promotes the progression of gastric cancer via AKT/mTOR and Wnt3a/β-catenin signaling pathways.","date":"2020","source":"Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie","url":"https://pubmed.ncbi.nlm.nih.gov/32570120","citation_count":18,"is_preprint":false},{"pmid":"25038457","id":"PMC_25038457","title":"Predicting and validating the pathway of Wnt3a-driven suppression of osteoclastogenesis.","date":"2014","source":"Cellular signalling","url":"https://pubmed.ncbi.nlm.nih.gov/25038457","citation_count":17,"is_preprint":false},{"pmid":"25651906","id":"PMC_25651906","title":"Wnt3a signal pathways activate MyoD expression by targeting cis-elements inside and outside its distal enhancer.","date":"2015","source":"Bioscience reports","url":"https://pubmed.ncbi.nlm.nih.gov/25651906","citation_count":17,"is_preprint":false},{"pmid":"28534940","id":"PMC_28534940","title":"The Wnt regulator SFRP4 inhibits mesothelioma cell proliferation, migration, and antagonizes Wnt3a via its netrin-like domain.","date":"2017","source":"International journal of oncology","url":"https://pubmed.ncbi.nlm.nih.gov/28534940","citation_count":17,"is_preprint":false},{"pmid":"27557659","id":"PMC_27557659","title":"Pigment Epithelium-derived Factor (PEDF) Blocks Wnt3a Protein-induced Autophagy in Pancreatic Intraepithelial Neoplasms.","date":"2016","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/27557659","citation_count":17,"is_preprint":false},{"pmid":"28901526","id":"PMC_28901526","title":"MicroRNA‑214 targets Wnt3a to suppress liver cancer cell proliferation.","date":"2017","source":"Molecular medicine reports","url":"https://pubmed.ncbi.nlm.nih.gov/28901526","citation_count":16,"is_preprint":false},{"pmid":"33190089","id":"PMC_33190089","title":"CBP-mediated Wnt3a/β-catenin signaling promotes cervical oncogenesis initiated by Piwil2.","date":"2020","source":"Neoplasia (New York, N.Y.)","url":"https://pubmed.ncbi.nlm.nih.gov/33190089","citation_count":15,"is_preprint":false},{"pmid":"32252152","id":"PMC_32252152","title":"Wnt3a and wnt5a as Potential Chondrogenic Stimulators for Nucleus Pulposus Cell Induction: A Comprehensive Review.","date":"2020","source":"Neurospine","url":"https://pubmed.ncbi.nlm.nih.gov/32252152","citation_count":15,"is_preprint":false},{"pmid":"36924567","id":"PMC_36924567","title":"Protective effects of oridonin against osteoporosis by regulating immunity and activating the Wnt3a/β-catenin/VEGF pathway in ovariectomized mice.","date":"2023","source":"International immunopharmacology","url":"https://pubmed.ncbi.nlm.nih.gov/36924567","citation_count":15,"is_preprint":false},{"pmid":"28587752","id":"PMC_28587752","title":"Dickkopf-1 inhibits Wnt3a-induced migration and epithelial-mesenchymal transition of human lens epithelial cells.","date":"2017","source":"Experimental eye research","url":"https://pubmed.ncbi.nlm.nih.gov/28587752","citation_count":15,"is_preprint":false},{"pmid":"28694190","id":"PMC_28694190","title":"Non-canonical pathway induced by Wnt3a regulates β-catenin via Pyk2 in differentiating human neural progenitor cells.","date":"2017","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/28694190","citation_count":15,"is_preprint":false},{"pmid":"28351228","id":"PMC_28351228","title":"PEGylated liposomes associate with Wnt3A protein and expand putative stem cells in human bone marrow populations.","date":"2017","source":"Nanomedicine (London, England)","url":"https://pubmed.ncbi.nlm.nih.gov/28351228","citation_count":15,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":54338,"output_tokens":8808,"usd":0.147567},"stage2":{"model":"claude-opus-4-6","input_tokens":12906,"output_tokens":4630,"usd":0.27042},"total_usd":0.417987,"stage1_batch_id":"msgbatch_01BparvgRZ9S2LMidvaoo86K","stage2_batch_id":"msgbatch_01NkBfv9SVZWthhS7j6mU6G3","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 1999,\n      \"finding\": \"Genetic epistasis shows that LEF-1 and TCF-1 transcription factors act downstream of WNT3A; double knockout of Lef1 and Tcf1 phenocopies Wnt3a-deficient mice (paraxial mesoderm defects, extra neural tubes, limb bud failure), demonstrating that LEF-1/TCF-1 are the primary nuclear effectors of WNT3A signaling in early mouse embryogenesis.\",\n      \"method\": \"Genetic double-knockout mouse (Lef1−/−Tcf1−/−) phenotypic analysis and epistasis\",\n      \"journal\": \"Genes & Development\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — reciprocal genetic epistasis in vivo; strong phenocopy evidence; replicated by independent labs studying Wnt3a\",\n      \"pmids\": [\"10090727\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1998,\n      \"finding\": \"Soluble WNT3A secreted from transfected L cells stabilizes β-catenin in recipient cells and reorganizes the actin cytoskeleton (directional alignment of stress fibers, redistribution of villin to leading edges) in C57MG mammary epithelial cells, establishing that WNT3A acts as a diffusible paracrine signal that both activates β-catenin and remodels cytoskeletal architecture.\",\n      \"method\": \"Conditioned medium transfer assay, immunostaining for actin/adhesion proteins, β-catenin Western blot\",\n      \"journal\": \"Genes to Cells\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — multiple orthogonal readouts (β-catenin stabilization, cytoskeletal reorganization) in defined cell-based assay; foundational paper\",\n      \"pmids\": [\"9893023\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2004,\n      \"finding\": \"WNT3A activates cell proliferation via two parallel pathways: (1) the canonical β-catenin/TCF-4 pathway and (2) a β-catenin-independent Raf-1–MEK–ERK cascade; ERK activation by WNT3A persists even when β-catenin is depleted by siRNA, and β-catenin can secondarily activate ERK through a TCF-4-dependent transcriptional event.\",\n      \"method\": \"siRNA knockdown of β-catenin and ERK, MEK inhibitor (U0126), dominant-negative TCF-4 transfection, proliferation and cell cycle assays in NIH3T3 cells\",\n      \"journal\": \"Journal of Cell Science\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — multiple orthogonal genetic and pharmacological perturbations; two independent pathway arms defined\",\n      \"pmids\": [\"15615777\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"WNT3A secretion requires PORCN-dependent lipid (palmitoyleoyl) modification at Ser209 of WNT3A, and the WNT3A–WLS complex accumulates when vacuolar acidification is inhibited; WLS is ER-resident in human cells and binds WNT3A through its lipid-binding domain, with vacuolar acidification needed to release palmitoylated WNT3A from WLS into secretory vesicles.\",\n      \"method\": \"V-ATPase inhibitor treatment, mutagenesis of Ser209, co-immunoprecipitation of WNT3A–WLS, cell fractionation, Xenopus convergent extension assay, structural modeling\",\n      \"journal\": \"Journal of Cell Science\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — mutagenesis of lipidation site combined with pharmacological dissection and co-IP; multiple orthogonal methods\",\n      \"pmids\": [\"20826466\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2005,\n      \"finding\": \"WNT3A acts as a long-range signaling molecule from the primitive streak/dorsal posterior node that activates the Delta/Notch pathway to regulate perinodal Nodal expression (left-right determination) while simultaneously controlling the segmentation clock oscillations of both Wnt/β-catenin and Notch pathways, genetically linking the segmentation clock to left-right axis specification.\",\n      \"method\": \"Wnt3a knockout mouse analysis, in situ hybridization, genetic epistasis with Notch/Delta pathway components\",\n      \"journal\": \"Development\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — in vivo knockout with defined molecular pathway placement; multiple target genes assessed\",\n      \"pmids\": [\"16291790\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"Integrin-linked kinase (ILK) activity is required for WNT3A-induced β-catenin stabilization and nuclear translocation; ILK inhibition suppresses GSK-3β Ser9 phosphorylation triggered by WNT3A conditioned medium and reverses β-catenin/LEF-mediated transcription; ILK co-immunoprecipitates with APC and GSK-3β as part of a Wnt pathway complex.\",\n      \"method\": \"Pharmacological ILK inhibition (QLT-0267), molecular inhibition constructs, purified WNT3A protein treatment, co-IP of ILK with APC/GSK-3β, β-catenin stabilization and nuclear translocation assays\",\n      \"journal\": \"Oncogene\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — reciprocal co-IP, pharmacological and molecular inhibition with multiple readouts; purified protein used to exclude secondary effects\",\n      \"pmids\": [\"16799642\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"WNT3A stimulation, via Frizzled and Dishevelled, activates PI4-kinase type IIα to produce PtdIns(4)P; Dvl directly interacts with and activates PI4KIIα (increasing its Vmax for ATP and PtdIns), and Dvl, PI4KIIα, and PIP5KI form a ternary complex upon WNT3A stimulation to efficiently produce PtdIns(4,5)P2 from PtdIns.\",\n      \"method\": \"Co-immunoprecipitation, in vitro kinase assay with purified components, lipid measurement in HEK293T cells, dominant-negative Fz and Dvl constructs\",\n      \"journal\": \"Journal of Biological Chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — direct binding and in vitro kinase activation assay combined with cellular co-IP; ternary complex reconstituted\",\n      \"pmids\": [\"19561074\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"WNT3A-stimulated RhoA GTPase activity is required for full induction of a subset of canonical WNT3A target genes in a β-catenin-dependent transcriptional program, but RhoA does not affect β-catenin stabilization or nuclear translocation itself; Rho activation is an independent co-requirement for WNT3A-driven osteoblastic differentiation.\",\n      \"method\": \"RhoA inhibition (C3 toxin, dominant-negative RhoA), microarray mRNA expression, real-time RT-PCR, C3H10T1/2 osteoblastic differentiation assay\",\n      \"journal\": \"Cellular Signalling\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — genome-wide transcriptomics plus pharmacological epistasis; single lab\",\n      \"pmids\": [\"19482078\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"WNT3A stimulation of mammalian Frizzled receptors triggers guanine-nucleotide exchange on heterotrimeric Go/Gi proteins; this response is blocked by pertussis toxin (which ADP-ribosylates Go/Gi) and by a Wnt antagonist, demonstrating that mammalian Frizzled receptors function as bona fide GPCRs coupling to Go/i upon WNT3A binding.\",\n      \"method\": \"GTPγS-binding assay on rat brain membranes and cultured cells, pertussis toxin treatment, Wnt antagonist blockade\",\n      \"journal\": \"Biochemical Journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — direct biochemical GEF assay on native membranes; pertussis toxin provides mechanistic specificity; multiple cellular contexts tested\",\n      \"pmids\": [\"21128903\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"WNT3A undergoes basolateral secretion in polarized epithelial cells via a clathrin/adaptor protein-1 (AP-1)-dependent route, and this requires WLS recycling by AP-2; WNT3A carries two high-mannose-type N-glycans (at Asn87 and Asn298), and glycosylation processing at these sites does not redirect WNT3A to apical secretion (unlike WNT11, which uses complex-type glycan at Asn40 for apical targeting).\",\n      \"method\": \"Mass spectrometric glycan analysis, clathrin/AP-1 knockdown, AP-2 disruption, polarized MDCK cell secretion assay, WLS localization by immunostaining\",\n      \"journal\": \"Journal of Cell Science\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — mass spectrometry for glycan identification combined with genetic knockdown of trafficking machinery; orthogonal readouts\",\n      \"pmids\": [\"23613470\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"WNT3A activates the PI3K–Akt pathway independently of β-catenin in NIH3T3 fibroblasts; Wnt3a-induced Akt activation is abrogated by PI3K inhibitors (LY294002, wortmannin) but not by MEK inhibitor U0126, and activated Akt transiently accumulates in the nucleus; PI3K–Akt activation contributes to WNT3A-induced cell proliferation.\",\n      \"method\": \"PI3K inhibitors, Akt siRNA, β-catenin siRNA, subcellular fractionation/nuclear Akt detection, proliferation assay in NIH3T3 cells\",\n      \"journal\": \"Cellular Signalling\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — pharmacological and siRNA epistasis; single lab but multiple perturbations\",\n      \"pmids\": [\"17011750\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"WNT3A induces GSK-3β Ser9 phosphorylation and β-catenin accumulation through activation of the RhoA/ROCK axis; shRNA knockdown of RhoA or ROCK inhibition (Y27632) abolishes both GSK-3β phosphorylation and β-catenin accumulation induced by WNT3A; purified active ROCK directly phosphorylates recombinant GSK-3β in vitro.\",\n      \"method\": \"shRNA against RhoA, ROCK inhibitor Y27632, Tat-C3 (RhoA inhibitor), in vitro kinase assay with purified ROCK and recombinant GSK-3β, Western blot in RAW264.7 and HEK293 cells\",\n      \"journal\": \"Journal of Cellular Physiology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — in vitro kinase reconstitution of ROCK→GSK-3β plus genetic (shRNA) and pharmacological epistasis\",\n      \"pmids\": [\"27575935\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"WNT3A promotes superoxide generation leading to Src-mediated Tyr42 phosphorylation of RhoA; phospho-Tyr42-RhoA binds to β-catenin via β-catenin's N-terminal domain and the complex translocates to the nucleus, where it associates with the Vimentin (Vim) gene promoter to drive vimentin transcription.\",\n      \"method\": \"Co-immunoprecipitation of p-Tyr42 RhoA with β-catenin, ChIP on Vim promoter, domain mapping (N-terminal β-catenin deletion), ROS detection, NADPH oxidase inhibition, ROCK2 inhibition\",\n      \"journal\": \"Redox Biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — reciprocal co-IP plus ChIP; single lab but multiple mechanistic readouts\",\n      \"pmids\": [\"33388549\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"Cryo-EM structure of human WLS in complex with WNT3A at 2.2 Å reveals that the WLS transmembrane domain adopts a GPCR-like fold with a conserved core cavity and lateral opening; the palmitoyleoyl lipid moiety on WNT3A (at its conserved palmitoleoylation site on a β-hairpin) threads through a hydrophobic tunnel of the WLS transmembrane domain and inserts into the membrane; this WNT3A β-hairpin–WLS interaction is essential for WLS-mediated Wnt secretion.\",\n      \"method\": \"Cryo-EM structure determination (2.2 Å resolution), functional validation of lipid-binding contacts, mutagenesis of WNT3A palmitoleoylation site\",\n      \"journal\": \"Nature Communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — high-resolution cryo-EM structure with mutagenesis validation; defines atomic mechanism of WNT3A–WLS interaction\",\n      \"pmids\": [\"34315898\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"WNT3A assembles into high-molecular-weight homo-trimeric and larger complexes in serum-containing media; binding to the extracellular CRD of Frizzled8 or to sFRP2 dissociates these complexes; assembly restricts WNT3A diffusion range in Xenopus embryos as shown by fluorescence correlation spectroscopy and immunohistochemistry.\",\n      \"method\": \"Analytical ultracentrifugation with fluorescence detection (AUC-FDS), gel filtration + cross-linking + single-particle analysis, fluorescence correlation spectroscopy (FCS), immunohistochemistry in Xenopus embryos\",\n      \"journal\": \"Communications Biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — multiple biophysical methods (AUC, FCS, single-particle EM) combined with in vivo diffusion measurement; mechanistically rigorous\",\n      \"pmids\": [\"30320232\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"Mechanical strain and WNT3A/β-catenin signaling synergize to drive cells through mitosis: strain alone induces Src-dependent Y654 phosphorylation of β-catenin and S/G2 accumulation without division; addition of WNT3A (or CKI inhibition) raises β-catenin-mediated transcription to a threshold required to trigger mitosis in strained cells.\",\n      \"method\": \"Mechanical strain device on MDCK cells, Casein Kinase I inhibitor, WNT3A addition, β-catenin Y654 phospho-specific antibody, cell cycle analysis, β-catenin reporter assay\",\n      \"journal\": \"eLife\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — quantitative mechanistic dissection combining mechanical and biochemical perturbations; single lab\",\n      \"pmids\": [\"27782880\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"PDGF receptor transactivation is the predominant β-catenin-independent mechanism by which WNT3A stimulates osteoblastic cell proliferation: WNT3A activates Src family kinases (SFKs) via Dishevelled, which transactivate PDGF receptors; soluble decoy PDGF-R excludes autocrine PDGF as the source; siRNA knockdown of PDGF-R confirms its predominant role.\",\n      \"method\": \"Selective kinase inhibitors (SFK, PDGF-R, PKC), DKK1 to exclude β-catenin contribution, PDGF-R siRNA, soluble decoy PDGF-R, time-course phosphorylation analysis in MC3T3-E1 cells\",\n      \"journal\": \"Journal of Bone and Mineral Research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — multiple pharmacological and genetic epistasis approaches; single lab but internally consistent\",\n      \"pmids\": [\"22927028\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"WNT3A transcriptionally induces Lef-1 gene expression in airway submucosal gland progenitor cells in vivo through TCF4/β-catenin binding to a Wnt-responsive region in the Lef-1 promoter; ChIP confirmed TCF4 occupancy, and Wnt3a-deficient mice show loss of Lef-1 induction in forming gland buds.\",\n      \"method\": \"ChIP for TCF4 on Lef-1 promoter in primary airway epithelial cells, Lef-1 promoter–GFP reporter mice crossed to Wnt3a-deficient mice, dominant-active β-catenin + TCF4 transfection luciferase assay\",\n      \"journal\": \"Developmental Biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — ChIP with in vivo genetic validation in Wnt3a KO mice; mechanistic target gene regulation established\",\n      \"pmids\": [\"17335794\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"Sox17 directly represses WNT3A/β-catenin-mediated transcriptional activation of the Lef-1 promoter; EMSA and ChIP identified four Sox17-binding sites in the Lef-1 promoter that collaborate with TCF4 sites; Sox17–TCF complexes form through protein–protein interaction via Sox17's DNA- or β-catenin-binding domains.\",\n      \"method\": \"EMSA, ChIP, site-directed mutagenesis of Sox17 and TCF binding sites, luciferase reporter assay in primary airway epithelial cells and cell lines\",\n      \"journal\": \"American Journal of Physiology – Lung Cellular and Molecular Physiology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — ChIP plus EMSA plus combinatorial site mutagenesis; single lab\",\n      \"pmids\": [\"20802155\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"WNT3A induces AChE expression in osteoblasts through a β-catenin–Runx2 axis; Wnt3a treatment increases Runx2 protein which binds a defined Runx2-binding site in the ACHE promoter (confirmed by ChIP), and deletion of this site reduces promoter activity; DKK-1 blocks the WNT3A-induced AChE expression.\",\n      \"method\": \"ChIP for Runx2 on ACHE promoter, ACHE promoter luciferase reporter with deletion mutants, Runx2 overexpression, DKK-1 inhibition, primary rat osteoblast culture\",\n      \"journal\": \"Journal of Biological Chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — ChIP plus promoter mutagenesis and functional rescue; single lab\",\n      \"pmids\": [\"28607150\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"LRP6, not LRP5, is the primary co-receptor mediating WNT3A canonical signaling in osteoblasts; genome-wide RNA-seq in Lrp5- and Lrp6-deficient primary calvarial osteoblasts treated with WNT3A identified 782 WNT3A-regulated genes, with LRP6 loss abolishing the majority of WNT3A transcriptional responses while LRP5 loss had minor effects.\",\n      \"method\": \"RNA-seq in Lrp5-KO and Lrp6-KO primary osteoblasts treated with WNT3A, receptor-specific genetic deletion\",\n      \"journal\": \"PLOS ONE\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — genome-wide transcriptomics with co-receptor-specific knockouts; single lab\",\n      \"pmids\": [\"29176883\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"WNT3A non-canonically triggers Ca2+ influx and CaMKII activation in differentiating human neural progenitor cells; elevated intracellular Ca2+ activates Pyk2, which in turn phosphorylates GSK-3β and stabilizes β-catenin, revealing a Wnt/Ca2+→Pyk2→GSK-3β→β-catenin cross-talk pathway.\",\n      \"method\": \"Intracellular Ca2+ imaging, CaMKII activity assay, Pyk2 phosphorylation Western blot, GSK-3β and β-catenin phosphorylation analysis in differentiating hNPCs\",\n      \"journal\": \"Biochemical and Biophysical Research Communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — multiple sequential pathway components measured; single lab\",\n      \"pmids\": [\"28694190\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"TLR4 activation increases Wnt3a and Dvl3 expression in monocytes; Wnt3a–Dvl3 signaling restrains TLR4-induced inflammatory responses by stabilizing β-catenin and suppressing NF-κB activity; siRNA or ectopic expression of Dvl3, GSK3β, and β-catenin confirmed the pathway; in vivo, Wnt3a inhibition during endotoxemia enhances systemic cytokines and neutrophil infiltration.\",\n      \"method\": \"siRNA knockdown and ectopic expression of Wnt3a, Dvl3, GSK3β, β-catenin in primary monocytes; ELISA, Western blot, NF-κB DNA binding assay; murine endotoxemia model with Wnt3a inhibition\",\n      \"journal\": \"Molecular Immunology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — gain- and loss-of-function in primary cells plus in vivo model; single lab\",\n      \"pmids\": [\"31884387\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"JAK3 phospho-inactivates the E3 ubiquitin ligase Nedd4-2, reducing ubiquitination of Wnt3 protein and thereby increasing Wnt3a protein levels; elevated Wnt3a then signals through Dvl3 and β-catenin to suppress NF-κB and pro-inflammatory cytokines in innate immune cells; JAK3 or Wnt3a inhibition in vivo worsens P. gingivalis-induced periodontal disease.\",\n      \"method\": \"Gain- and loss-of-function of JAK3, Nedd4-2 ubiquitination assay, Wnt3 co-immunoprecipitation with Nedd4-2, Dvl3 and β-catenin downstream analysis, in vivo periodontal disease mouse model\",\n      \"journal\": \"FASEB Journal\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — ubiquitination assay plus co-IP plus in vivo model; single lab but multiple mechanistic steps defined\",\n      \"pmids\": [\"32433819\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"WNT3A activates MyoD expression by directly targeting cis-regulatory elements: WNT3A highly activates a composite proximal promoter + distal enhancer reporter and targets an upstream −8 to −9 kb Wnt-response element (L fragment) through Pax3/Pax7-binding sites; β-catenin and Pax7 binding to their respective sites in the distal enhancer and L fragment was confirmed by ChIP.\",\n      \"method\": \"Luciferase reporter deletion analysis of MyoD cis-elements, ChIP for β-catenin and Pax7, dominant-negative pathway constructs in HEK293T cells\",\n      \"journal\": \"Bioscience Reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — ChIP plus systematic promoter/enhancer deletion mapping; single lab\",\n      \"pmids\": [\"25651906\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"Intranasal WNT3A activates the Frizzled-1/PIWIL1a/FOXM1 pathway to reduce neuronal apoptosis after ischemic stroke; Frizzled-1 siRNA and PIWI1a siRNA reverse WNT3A neuroprotection, while PIWI1a CRISPR restoration after Frizzled-1 knockdown rescues FOXM1 and reduces cleaved caspase-3, placing Frizzled-1 upstream of PIWIL1a and FOXM1.\",\n      \"method\": \"Intranasal WNT3A administration in MCAO rats, Frizzled-1 siRNA, PIWI1a siRNA, PIWI1a CRISPR restoration, Western blot, immunohistochemistry, neurobehavioral testing, infarct volume measurement\",\n      \"journal\": \"Journal of Neuroscience\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — genetic epistasis (siRNA + CRISPR rescue) with defined pathway order; single lab\",\n      \"pmids\": [\"29954850\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"Rmb WNT3A normalizes altered hepatic lipogenesis and cholesterol biosynthesis in LRP6(R611C) mutant mice by reversing the Sp1-dependent IGF1/AKT/mTORC1/mTORC2 signaling activation that causes combined hyperlipidemia; in vitro rmWNT3A treatment of LRP6-mutant hepatocytes also restores pathway signaling to normal.\",\n      \"method\": \"In vivo rmWNT3A administration to LRP6(R611C) mice, in vitro primary hepatocyte treatment, IGF1R antagonist and rapamycin comparisons, plasma lipid and gene expression analysis\",\n      \"journal\": \"Cell Metabolism\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — in vivo and in vitro rescue with rmWNT3A; pathway normalization validated by multiple enzyme expression readouts; single lab\",\n      \"pmids\": [\"24506864\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"Localized WNT3A signaling induces asymmetric division of mouse embryonic stem cells; H3K27me3 (but not H3K4me3) modifications are correlated with gene expression changes that determine daughter cell fate after WNT3A-induced asymmetric division, as revealed by joint single-cell epigenome+transcriptome profiling.\",\n      \"method\": \"Same-cell epigenome and transcriptome sequencing (H3K27me3/H3K4me3 + gene expression), localized WNT3A stimulation of mouse ESCs\",\n      \"journal\": \"Nature Communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — novel single-cell multiomic method applied to mechanistic question; single lab\",\n      \"pmids\": [\"34642323\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"WNT3A-loaded exosomes activate canonical WNT signaling in intact cartilage in vivo (where soluble recombinant WNT3A fails), and a single injection improves osteochondral defect repair in mice over 8 weeks, demonstrating that exosomal delivery overcomes the hydrophobic barrier to WNT3A tissue penetration.\",\n      \"method\": \"TOPFlash reporter assay for in vitro WNT activity, TCF/Lef:H2B-GFP reporter mice for in vivo signaling, osteochondral defect mouse model, histological scoring\",\n      \"journal\": \"Journal of Extracellular Vesicles\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — in vitro and in vivo comparison of soluble vs. exosomal WNT3A with pathway reporter; single lab\",\n      \"pmids\": [\"34025953\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"WNT3A protein requires association with a lipid vesicle for in vivo stability and activity; liposome-reconstituted WNT3A (L-WNT3A) associates with the external lipid membrane surface, prolonging pathway activation in primary cells, and activates mesenchymal/skeletal stem cell populations in bone marrow engraftment assays in vivo.\",\n      \"method\": \"Liposome reconstitution of WNT3A, primary cell Wnt reporter assays, bone marrow engraftment in vivo assay, stem cell population analysis\",\n      \"journal\": \"PLOS ONE\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — biophysical protein-lipid association combined with in vivo functional validation; single lab\",\n      \"pmids\": [\"24400074\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"Nintedanib inhibits WNT3A-induced myofibroblast activation by blocking Src kinase activation and Src-dependent Y654 phosphorylation of β-catenin, thereby preventing β-catenin nuclear translocation; Src knockdown phenocopies nintedanib treatment, placing Src downstream of WNT3A and upstream of β-catenin nuclear entry in pulmonary fibroblasts.\",\n      \"method\": \"Nintedanib treatment, Src-specific siRNA knockdown, Src inhibitor KX2-391, β-catenin Y654 phosphorylation Western blot, nuclear fractionation, downstream Wnt target gene expression in lung fibroblasts\",\n      \"journal\": \"Frontiers in Pharmacology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — pharmacological and genetic (siRNA) epistasis; single lab but multiple orthogonal approaches\",\n      \"pmids\": [\"32231574\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"Heparanase modulates WNT3A signaling in medulloblastoma cells by altering syndecan co-receptor function and GEF-H1 (a Rho-GEF) distribution; HPSE pretreatment differentially regulates Rac1/RhoA activities after WNT3A exposure, and the HPSE/HSPG axis modulates WNT3A-dependent β-catenin and N-Myc distribution as well as Gli transcription factor gene expression.\",\n      \"method\": \"Exogenous active heparanase treatment, Rac1/RhoA activity assays, immunofluorescence for GEF-H1/β-catenin, RT-PCR for Wnt target and HSPG genes, MB cell proliferation and invasion assays\",\n      \"journal\": \"Experimental and Therapeutic Medicine\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 — single lab, correlative mechanistic data without direct binding/reconstitution of WNT3A–HSPG interaction\",\n      \"pmids\": [\"21442027\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"WNT3A is a palmitoyleoylated (Ser209), N-glycosylated secreted ligand that binds WLS via its lipid moiety within a hydrophobic transmembrane tunnel (cryo-EM structure resolved at 2.2 Å); vacuolar acidification releases WNT3A from the WLS complex for basolateral, clathrin/AP-1-dependent secretion, after which WNT3A assembles into diffusion-restricted homo-trimeric complexes that are dissociated by Frizzled-CRD or sFRP binding; upon receptor engagement, WNT3A acts as a bona fide GPCR ligand for Frizzled/Go/i proteins and signals through canonical β-catenin stabilization (requiring LEF-1/TCF nuclear effectors, and facilitated by ILK, RhoA/ROCK-mediated GSK-3β phosphorylation, and Pyk2-mediated Ca2+-dependent cross-talk), as well as β-catenin-independent activation of Raf-MEK-ERK, PI3K-Akt, RhoA/ROCK, and phosphoinositide (PI4K/PIP5K) pathways, collectively regulating paraxial mesoderm formation, left-right axis determination, segmentation, neural progenitor proliferation, osteoblast differentiation, and immune modulation.\"\n}\n```","stage2_raw":"```json\n{\n  \"mechanistic_narrative\": \"WNT3A is a palmitoyleoylated, N-glycosylated secreted ligand that activates both canonical β-catenin and multiple β-catenin-independent signaling cascades to regulate paraxial mesoderm formation, left-right axis determination, somite segmentation, osteoblast differentiation, neural progenitor proliferation, and innate immune modulation [PMID:10090727, PMID:16291790, PMID:19482078, PMID:31884387]. The lipid moiety at Ser209, added by PORCN, threads through a hydrophobic tunnel in the WLS transmembrane domain (resolved at 2.2 Å by cryo-EM) to mediate ER-to-plasma-membrane trafficking via clathrin/AP-1-dependent basolateral secretion; extracellularly, WNT3A assembles into diffusion-restricted homo-trimeric complexes that are dissociated by Frizzled-CRD or sFRP binding [PMID:20826466, PMID:34315898, PMID:23613470, PMID:30320232]. Upon receptor engagement, WNT3A functions as a GPCR ligand coupling Frizzled to Go/Gi proteins and signals through LRP6-dependent β-catenin stabilization (requiring LEF-1/TCF nuclear effectors and facilitated by ILK, RhoA/ROCK-mediated GSK-3β phosphorylation, and Pyk2-mediated Ca²⁺ cross-talk), as well as β-catenin-independent Raf-MEK-ERK, PI3K-Akt, Src-PDGFR transactivation, and PI4KIIα/PIP5KI phosphoinositide pathways [PMID:21128903, PMID:15615777, PMID:16799642, PMID:27575935, PMID:28694190, PMID:19561074, PMID:22927028, PMID:29176883]. WNT3A-driven β-catenin transcriptional activity, modulated by H3K27me3 epigenetic marks, also directs asymmetric stem cell division and cooperates with mechanical strain to trigger mitosis [PMID:34642323, PMID:27782880].\",\n  \"teleology\": [\n    {\n      \"year\": 1998,\n      \"claim\": \"Establishing that WNT3A functions as a diffusible paracrine signal: conditioned medium transfer showed that secreted WNT3A stabilizes β-catenin and remodels the actin cytoskeleton in recipient cells, providing the first demonstration that WNT3A has both canonical and cytoskeletal outputs.\",\n      \"evidence\": \"Conditioned medium from WNT3A-transfected L cells applied to C57MG cells; β-catenin Western blot and actin/adhesion protein immunostaining\",\n      \"pmids\": [\"9893023\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Receptor identity for WNT3A not yet determined\", \"Mechanism of cytoskeletal reorganization downstream of WNT3A undefined\", \"Whether WNT3A acts at long range in vivo unknown\"]\n    },\n    {\n      \"year\": 1999,\n      \"claim\": \"Identifying the nuclear effectors: genetic epistasis showed that LEF-1 and TCF-1 double knockout phenocopies Wnt3a deficiency in mice, establishing them as essential downstream transcription factors for WNT3A in paraxial mesoderm and neural tube patterning.\",\n      \"evidence\": \"Lef1−/−;Tcf1−/− double-knockout mouse phenotypic comparison with Wnt3a−/− mice\",\n      \"pmids\": [\"10090727\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether additional TCF family members contribute in other tissues unknown\", \"Signal transduction steps between WNT3A and LEF-1/TCF nuclear activation undefined\"]\n    },\n    {\n      \"year\": 2004,\n      \"claim\": \"Resolving pathway bifurcation: WNT3A was shown to activate cell proliferation through two parallel arms — canonical β-catenin/TCF-4 and a β-catenin-independent Raf-1–MEK–ERK cascade — demonstrating that WNT3A is not solely a canonical Wnt ligand.\",\n      \"evidence\": \"β-catenin siRNA, MEK inhibitor U0126, dominant-negative TCF-4 in NIH3T3 cells with proliferation readouts\",\n      \"pmids\": [\"15615777\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Proximal receptor-level mechanism activating ERK independently of β-catenin not identified\", \"Whether the two arms converge on shared gene targets unclear\"]\n    },\n    {\n      \"year\": 2005,\n      \"claim\": \"Linking WNT3A to developmental patterning beyond mesoderm: Wnt3a knockout mice revealed that WNT3A from the primitive streak/dorsal node activates Delta/Notch to regulate perinodal Nodal expression for left-right axis determination while simultaneously controlling the segmentation clock.\",\n      \"evidence\": \"Wnt3a knockout mouse with in situ hybridization and genetic epistasis with Notch/Delta components\",\n      \"pmids\": [\"16291790\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Direct versus indirect activation of Delta/Notch by WNT3A not resolved\", \"Whether WNT3A protein acts at long range to reach perinodal cells or requires relay unknown\"]\n    },\n    {\n      \"year\": 2006,\n      \"claim\": \"Expanding the non-canonical signaling repertoire: WNT3A was found to activate PI3K–Akt independently of β-catenin and to require ILK for GSK-3β Ser9 phosphorylation and β-catenin stabilization, adding two new signaling branches to the WNT3A network.\",\n      \"evidence\": \"PI3K inhibitors and Akt siRNA in NIH3T3 cells; ILK pharmacological inhibition plus co-IP of ILK with APC/GSK-3β\",\n      \"pmids\": [\"17011750\", \"16799642\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How ILK is activated by WNT3A receptor complex unknown\", \"Whether PI3K–Akt and ILK pathways cross-talk in WNT3A context untested\"]\n    },\n    {\n      \"year\": 2009,\n      \"claim\": \"Defining phosphoinositide and RhoA co-requirement: WNT3A was shown to activate PI4KIIα through Dvl to produce PtdIns(4,5)P₂ via a ternary Dvl–PI4KIIα–PIP5KI complex, and separately RhoA GTPase activity was found to be required for a subset of β-catenin-dependent WNT3A target genes and osteoblast differentiation.\",\n      \"evidence\": \"In vitro kinase reconstitution with purified PI4KIIα and Dvl; RhoA inhibition (C3 toxin) with microarray in C3H10T1/2 cells\",\n      \"pmids\": [\"19561074\", \"19482078\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How Dvl is selectively directed to PI4KIIα versus other effectors unknown\", \"Whether RhoA-dependent gene subset is tissue-specific untested\"]\n    },\n    {\n      \"year\": 2010,\n      \"claim\": \"Elucidating the secretory mechanism: PORCN-dependent palmitoyleoylation at Ser209 was shown to be required for WNT3A–WLS binding, and vacuolar acidification was found necessary to release WNT3A from WLS into secretory vesicles, establishing the ER-to-vesicle transit mechanism.\",\n      \"evidence\": \"Ser209 mutagenesis, V-ATPase inhibitor treatment, WNT3A–WLS co-immunoprecipitation, cell fractionation\",\n      \"pmids\": [\"20826466\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Structural basis of pH-dependent WNT3A–WLS dissociation unknown at this stage\", \"Post-WLS carriers of WNT3A not identified\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Establishing Frizzled as a bona fide GPCR for WNT3A: direct GTPγS-binding assays demonstrated that WNT3A triggers Frizzled-mediated guanine-nucleotide exchange on Go/Gi proteins, blocked by pertussis toxin, proving GPCR-like signaling.\",\n      \"evidence\": \"GTPγS-binding assay on rat brain membranes and cultured cells with pertussis toxin and Wnt antagonist\",\n      \"pmids\": [\"21128903\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Which specific Frizzled family member(s) couple to Go/Gi upon WNT3A not resolved\", \"Downstream effectors of Go/Gi in this context not identified\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Defining polarized secretion and receptor transactivation: WNT3A was shown to undergo basolateral secretion via clathrin/AP-1 with two high-mannose N-glycans, and separately WNT3A was found to stimulate osteoblast proliferation through Src/Dishevelled-mediated PDGFR transactivation independent of β-catenin.\",\n      \"evidence\": \"Mass spectrometry glycan analysis with AP-1/AP-2 knockdown in MDCK cells; PDGFR siRNA and Src inhibitors in MC3T3-E1 osteoblasts\",\n      \"pmids\": [\"23613470\", \"22927028\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether basolateral polarity of WNT3A secretion is universal across epithelia unknown\", \"PDGFR transactivation mechanism (direct phosphorylation vs. adaptor) not molecularly defined\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Reconstituting the RhoA/ROCK–GSK-3β axis and mechanical synergy: in vitro kinase assays showed ROCK directly phosphorylates GSK-3β Ser9, establishing a RhoA/ROCK branch that feeds into canonical β-catenin stabilization; concurrently, mechanical strain was shown to synergize with WNT3A to drive mitosis via a β-catenin transcriptional threshold mechanism.\",\n      \"evidence\": \"Purified ROCK kinase assay on recombinant GSK-3β with shRNA validation; mechanical strain device on MDCK cells with WNT3A addition and cell cycle analysis\",\n      \"pmids\": [\"27575935\", \"27782880\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether ROCK-mediated GSK-3β phosphorylation operates in all WNT3A-responsive tissues untested\", \"How mechanical strain integrates with WNT3A at the receptor level unknown\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Clarifying co-receptor specificity and Ca²⁺ cross-talk: genome-wide RNA-seq in Lrp5/Lrp6-knockout osteoblasts revealed LRP6 as the primary canonical co-receptor for WNT3A, and in neural progenitors WNT3A was found to trigger Ca²⁺ influx activating Pyk2, which phosphorylates GSK-3β to stabilize β-catenin, bridging non-canonical Ca²⁺ and canonical pathways.\",\n      \"evidence\": \"RNA-seq in Lrp5-KO and Lrp6-KO primary osteoblasts; Ca²⁺ imaging and Pyk2 phosphorylation analysis in human neural progenitor cells\",\n      \"pmids\": [\"29176883\", \"28694190\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether LRP6 preference over LRP5 extends beyond osteoblasts unknown\", \"Ca²⁺ channel identity activated by WNT3A not identified\", \"Whether Pyk2-mediated cross-talk operates in non-neural cell types untested\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Revealing oligomeric state and ROS-dependent nuclear RhoA–β-catenin complex: biophysical analyses demonstrated WNT3A forms homo-trimeric complexes that restrict diffusion and are dissociated by Frizzled-CRD or sFRP binding; separately, WNT3A-induced superoxide was shown to drive Src-mediated Tyr42 phosphorylation of RhoA, enabling RhoA–β-catenin nuclear translocation to the Vimentin promoter.\",\n      \"evidence\": \"Analytical ultracentrifugation, fluorescence correlation spectroscopy in Xenopus embryos; co-IP of p-Tyr42 RhoA with β-catenin and ChIP on Vim promoter\",\n      \"pmids\": [\"30320232\", \"33388549\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Stoichiometry and lipid requirements for WNT3A trimerization not fully defined\", \"Whether ROS-dependent RhoA–β-catenin complex is a general WNT3A mechanism or context-specific unknown\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Defining WNT3A's anti-inflammatory role: WNT3A/Dvl3/β-catenin signaling was found to restrain TLR4-induced NF-κB activation in monocytes, with JAK3-mediated stabilization of Wnt3 protein (via Nedd4-2 inactivation) amplifying this pathway; in vivo Wnt3a inhibition exacerbated endotoxemia and periodontal disease.\",\n      \"evidence\": \"siRNA/ectopic expression of Wnt3a, Dvl3, β-catenin in primary monocytes; JAK3–Nedd4-2 ubiquitination assays; murine endotoxemia and periodontal disease models\",\n      \"pmids\": [\"31884387\", \"32433819\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether WNT3A anti-inflammatory function is Frizzled subtype-specific untested\", \"Direct binding of Nedd4-2 to WNT3A protein not demonstrated\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Atomic resolution of the WNT3A–WLS complex and epigenetic outputs: cryo-EM at 2.2 Å revealed the palmitoyleoyl lipid threading through a GPCR-like WLS transmembrane tunnel, establishing the structural basis of WNT secretion; concurrently, single-cell multi-omic profiling showed WNT3A-driven asymmetric stem cell division correlates with H3K27me3 redistribution.\",\n      \"evidence\": \"Cryo-EM structure with mutagenesis validation; same-cell epigenome+transcriptome sequencing in mouse ESCs\",\n      \"pmids\": [\"34315898\", \"34642323\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Structural basis of pH-dependent WNT3A release from WLS not captured in current structure\", \"Whether H3K27me3 changes are directly instructed by β-catenin/TCF or secondary unknown\", \"Structure of WNT3A bound to Frizzled CRD together with LRP6 not available\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"Major unresolved questions include: the structural basis of WNT3A engagement with the Frizzled–LRP6 holoreceptor complex, the mechanism by which Frizzled–Go/Gi coupling is selected over β-catenin-dependent versus -independent outputs, how WNT3A homo-trimers are formed and regulated in vivo, and whether the multiple β-catenin-independent branches (ERK, PI3K-Akt, RhoA, PDGFR transactivation, Ca²⁺/Pyk2) operate simultaneously or are context-selectively activated.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No ternary WNT3A–Frizzled–LRP6 structure exists\", \"Selectivity mechanism for canonical versus non-canonical pathway activation undefined\", \"In vivo regulation of WNT3A oligomeric state not established\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0048018\", \"supporting_discovery_ids\": [1, 8, 14]},\n      {\"term_id\": \"GO:0060089\", \"supporting_discovery_ids\": [8]},\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [2, 7, 11, 22]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005576\", \"supporting_discovery_ids\": [1, 14, 28, 29]},\n      {\"term_id\": \"GO:0005783\", \"supporting_discovery_ids\": [3, 13]},\n      {\"term_id\": \"GO:0031410\", \"supporting_discovery_ids\": [9, 28]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [0, 1, 2, 5, 6, 8, 10, 11, 21]},\n      {\"term_id\": \"R-HSA-1266738\", \"supporting_discovery_ids\": [0, 4, 24, 27]},\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [22, 23]},\n      {\"term_id\": \"R-HSA-9609507\", \"supporting_discovery_ids\": [3, 9, 13]}\n    ],\n    \"complexes\": [\n      \"WNT3A–WLS secretory complex\",\n      \"WNT3A homo-trimer\"\n    ],\n    \"partners\": [\n      \"WLS\",\n      \"LRP6\",\n      \"FZD1\",\n      \"FZD8\",\n      \"DVL3\",\n      \"ILK\",\n      \"SFRP2\"\n    ],\n    \"other_free_text\": []\n  }\n}\n```"}