{"gene":"WNT6","run_date":"2026-04-28T23:00:23","timeline":{"discoveries":[{"year":2011,"finding":"Wnt6, Wnt10a, and Wnt10b inhibit adipogenesis and stimulate osteoblastogenesis through a β-catenin-dependent mechanism; knockdown of β-catenin completely prevents these effects, establishing that downstream mechanisms of β-catenin are required for Wnt6 to influence mesenchymal stem cell fate.","method":"Gain- and loss-of-function in ST2 and 3T3-L1 cells; β-catenin knockdown epistasis; β-catenin stabilization assays","journal":"Bone","confidence":"High","confidence_rationale":"Tier 2 — reciprocal gain/loss-of-function with epistasis (β-catenin KD), replicated across two cell lines","pmids":["21872687"],"is_preprint":false},{"year":2001,"finding":"Human WNT6 encodes a 365-amino-acid secreted glycoprotein with an N-terminal signal peptide, WNT core domain, and RGD motif; the WNT6 and WNT10A genes are clustered head-to-tail within 7 kb on chromosome 2q35.","method":"Molecular cloning, sequence analysis, Northern blot expression analysis","journal":"Biochemical and biophysical research communications","confidence":"Medium","confidence_rationale":"Tier 2 — direct cloning and structural characterization, single lab","pmids":["11350055"],"is_preprint":false},{"year":2002,"finding":"Wnt-6 expressed in the ureter bud induces kidney tubulogenesis in vitro, rescues tubulogenesis in Wnt-4 mutant embryonic mesenchyme, and activates Wnt-4 transcription, placing Wnt-6 upstream of Wnt-4 in the tubulogenesis pathway; Wnt-6 also induces a secondary axis in Xenopus embryos, consistent with canonical pathway activation.","method":"Cell lines expressing Wnt-6 co-cultured with embryonic kidney mesenchyme; Wnt-4 mutant rescue assay; Xenopus secondary axis assay; in situ hybridization for tubulogenesis markers","journal":"Genesis","confidence":"High","confidence_rationale":"Tier 2 — functional rescue in mutant background plus multiple orthogonal assays (tubulogenesis, marker induction, secondary axis)","pmids":["11948913"],"is_preprint":false},{"year":2007,"finding":"Wnt6 induces specification and epithelialization of F9 embryonal carcinoma cells to primitive endoderm via a canonical Wnt6–β-catenin–Snail1 signaling cascade, involving nuclear translocation of β-catenin and Snail1 and appearance of cytokeratin intermediate filaments.","method":"Wnt6 conditioned medium treatment; Wnt6-expressing cell lines; LiCl GSK-3 inhibition; immunofluorescence for β-catenin and Snail1 nuclear translocation; cytokeratin staining; PKA activation for parietal endoderm EMT","journal":"Cellular signalling","confidence":"High","confidence_rationale":"Tier 2 — multiple orthogonal assays (conditioned medium, overexpression, pharmacological) with defined molecular pathway","pmids":["18160257"],"is_preprint":false},{"year":2007,"finding":"In chick embryos, Wnt6 derived from the surface ectoderm is necessary for neural crest induction and acts through the non-canonical (not β-catenin-dependent) signaling pathway; canonical Wnt signaling inhibited neural crest production in chick.","method":"In ovo Wnt6 loss-of-function and gain-of-function; canonical vs. non-canonical pathway reporter assays in chick embryos","journal":"Developmental dynamics","confidence":"High","confidence_rationale":"Tier 2 — loss-of-function with specific neural crest phenotype and pathway discrimination by reporter assays","pmids":["17685490"],"is_preprint":false},{"year":2008,"finding":"Wnt6 in Xenopus is required during organogenesis to restrict heart muscle development; Wnt6 loss-of-function results in abnormally large heart muscle, and this repressive role is mediated via repression of cardiogenic transcription factors (including GATA), since reinstating GATA function rescues cardiomyogenic differentiation.","method":"Xenopus loss-of-function (morpholinos); gain-of-function with activated canonical Wnt/β-catenin; GATA rescue epistasis; analysis of cardiogenic transcription factors and differentiation genes","journal":"Developmental biology","confidence":"High","confidence_rationale":"Tier 2 — loss-of-function with defined phenotype, epistasis with GATA rescue, gain-of-function validation","pmids":["18804460"],"is_preprint":false},{"year":2005,"finding":"Ectodermal Wnt-6 promotes limb myogenesis through a Pax3/Paraxis–Myf5 pathway in chick; ectopic Wnt-6 expression extends domains of Pax3, Paraxis, Myf5, Myogenin, Desmin, and MyHC while inhibiting MyoD, and rescues myogenic gene expression after ectoderm removal, identifying a distinct Wnt-6 pathway separate from a MyoD-dependent ectodermal signal.","method":"Ectopic Wnt-6 expression in chick limb bud in ovo; ectoderm removal; in situ hybridization for myogenic markers","journal":"Developmental biology","confidence":"High","confidence_rationale":"Tier 2 — gain-of-function and ectoderm ablation rescue with multiple marker readouts in vivo","pmids":["16271265"],"is_preprint":false},{"year":2006,"finding":"Neural tube-derived Wnt1 and Wnt3a inhibit Wnt6 expression in the medial surface ectoderm via dermomyotomal Wnt11; Wnt11 was identified as an epithelialization factor acting on the medial dermomyotome, placing Wnt6 and Wnt11 as factors maintaining the epithelial nature of the ventrolateral and dorsomedial dermomyotomal lips, respectively.","method":"Interference with neighboring signaling pathways in chick embryo; in ovo gain/loss-of-function; in situ hybridization","journal":"Development","confidence":"Medium","confidence_rationale":"Tier 2 — epistasis pathway placement in vivo, single lab","pmids":["16818447"],"is_preprint":false},{"year":2010,"finding":"Ectodermal Wnt6 acts as a negative regulator of limb chondrogenesis in the chick embryo; ectoderm ablation leads to ectopic Sox9 and Collagen II expression, and Wnt6 overexpression reduces Sox9 and Collagen II, placing Wnt6 upstream of Sox9 in the chondrogenic cascade.","method":"Ectoderm ablation in chick limb bud; ectopic Wnt6 expression; in situ hybridization for Sox9, Collagen II","journal":"BMC developmental biology","confidence":"High","confidence_rationale":"Tier 2 — reciprocal gain/loss-of-function with consistent upstream pathway placement, in vivo","pmids":["20334703"],"is_preprint":false},{"year":2013,"finding":"Wnt6 is essential for uterine stromal cell proliferation during decidualization in mice; Wnt6 deficiency impairs stromal cell proliferation by downregulating cyclin B1 expression and prolonging cell cycle length, without adversely affecting decidual polyploidization.","method":"Wnt6-mutant mouse analysis; primary stromal cell culture; cell cycle analysis; cyclin B1 expression assay","journal":"Biology of reproduction","confidence":"High","confidence_rationale":"Tier 2 — genetic KO with defined molecular mechanism (cyclin B1) and specific cellular phenotype","pmids":["23175771"],"is_preprint":false},{"year":2013,"finding":"Wnt6 induces macrophage proliferation via a pertussis toxin-sensitive, ERK-mediated, β-catenin-independent induction of c-Myc; Wnt6 also drives macrophage polarization toward an M2-like phenotype by inducing Arginase-1 and downregulating TNF-α; Wnt6 expression in macrophages is induced via a TLR–MyD88–NF-κB-dependent pathway.","method":"Wnt6 conditioned medium treatment of macrophages; Wnt6-deficient macrophage analysis; thymidine incorporation proliferation assay; Ki-67 expression; pertussis toxin inhibition; ERK pathway analysis; Arginase-1 and TNF-α measurement","journal":"Journal of immunology","confidence":"High","confidence_rationale":"Tier 2 — loss-of-function, pharmacological inhibition (pertussis toxin), multiple orthogonal pathway readouts","pmids":["24123681"],"is_preprint":false},{"year":2021,"finding":"WNT6 promotes foam cell formation in macrophages by regulating acetyl-CoA carboxylase 2 (ACC2) and triacylglycerol (TAG) storage; genetic or pharmacological loss of WNT6 or ACC2 significantly reduced intracellular TAG levels and Mycobacterium tuberculosis survival in macrophages.","method":"Genetic (Wnt6-deficient macrophages) and pharmacological (ACC2 inhibitor) approaches; intracellular TAG measurement; Mtb survival assay; in vivo mouse TB model","journal":"Journal of Clinical Investigation","confidence":"High","confidence_rationale":"Tier 1–2 — genetic KO plus pharmacological inhibition with defined lipid metabolic mechanism and in vivo validation","pmids":["34255743"],"is_preprint":false},{"year":2015,"finding":"Sfrp2 binds Wnt6 and inhibits Wnt6 canonical signaling in cardiac progenitor cells (CPCs), which is required for inhibition of CPC proliferation; this canonical Wnt6 inhibition by Sfrp2 activates the non-canonical Wnt/PCP pathway through JNK, inducing cardiac transcription factors and CPC differentiation.","method":"Sfrp2-Wnt6 binding assay; canonical Wnt pathway reporter; JNK activation assay; CPC proliferation and differentiation assays; loss-of-function experiments","journal":"Journal of molecular and cellular cardiology","confidence":"High","confidence_rationale":"Tier 2 — direct binding demonstrated, pathway switching (canonical to non-canonical) with functional readouts","pmids":["26071893"],"is_preprint":false},{"year":2016,"finding":"Wnt6 signals through the canonical pathway in renal epithelial cells via the FZD7 receptor, as evidenced by increased GSK3β phosphorylation (Ser9), nuclear accumulation of β-catenin, and increased TCF/Lef transcriptional activity; Wnt6 also inhibits TGF-β-mediated p65-NF-κB nuclear translocation and rescues epithelial dedifferentiation.","method":"TCF/Lef reporter assay; GSK3β phosphorylation Western blot; β-catenin nuclear fractionation; 3D tubulogenesis assay; TGF-β rescue experiments; p65-NF-κB localization; p65(-/-) and IKKα/β(-/-) fibroblast experiments","journal":"American journal of physiology. Renal physiology","confidence":"High","confidence_rationale":"Tier 2 — multiple orthogonal assays identifying receptor (FZD7), downstream signaling, and functional consequence","pmids":["27122540"],"is_preprint":false},{"year":2009,"finding":"Wnt6 overexpression in human dental papilla cells enhances odontoblast-like differentiation (increased ALPase activity, upregulation of ALP, Col I, ON, OPN, BSP, DMP-1 gene expression, and mineral deposition) without significant effect on cell proliferation.","method":"Recombinant adenovirus Wnt6 overexpression; BrdU incorporation assay; flow cytometry; ALPase activity assay; mineralization assay; gene expression analysis","journal":"Journal of endodontics","confidence":"Medium","confidence_rationale":"Tier 2 — gain-of-function with multiple differentiation markers, single lab","pmids":["20113781"],"is_preprint":false},{"year":2014,"finding":"WNT6 activates the JNK (c-Jun N-terminal kinase) non-canonical signaling pathway in human dental pulp cells, promoting cell migration and odontoblast-like differentiation; blockage of JNK with SP600125 decreases but does not completely abolish WNT6-induced migration and differentiation, indicating additional pathway involvement.","method":"WNT6 conditioned medium treatment; JNK inhibitor SP600125; Western blot for JNK activation; RT-PCR for c-Jun; wound healing and transwell migration assays; ALP and Alizarin red staining; odontogenic gene expression","journal":"Journal of endodontics","confidence":"Medium","confidence_rationale":"Tier 2 — pharmacological inhibition with multiple functional readouts, single lab","pmids":["24935540"],"is_preprint":false},{"year":2012,"finding":"GATA6 and FOXA2 transcription factors directly bind the Wnt6 promoter and are sufficient to activate Wnt6 gene expression, leading to canonical WNT-β-catenin pathway activation and primitive endoderm induction in F9 cells.","method":"Chromatin immunoprecipitation (ChIP); Wnt6 promoter reporter assay; GATA6/FOXA2 overexpression; TCF-dependent transcription assay","journal":"Stem cells and development","confidence":"High","confidence_rationale":"Tier 1–2 — ChIP demonstrating direct promoter binding plus reporter and overexpression validation","pmids":["22607194"],"is_preprint":false},{"year":2009,"finding":"Coordinate Gα13–RhoA and Wnt6–β-catenin signaling are both required for retinoic acid-induced differentiation of F9 cells to primitive endoderm; Gα13/p115RhoGEF (but not RhoA alone) causes β-catenin nuclear translocation, and two independent pathways (canonical Wnt-β-catenin and Gα13-ERM) must both be active for primitive endoderm formation.","method":"Constitutively active Gα13(Q226L), p115RhoGEF, and RhoA(L63) overexpression; dominant-negative constructs; ROCK inhibitor Y-27632; β-catenin nuclear localization assay; F9 differentiation markers","journal":"Biochemistry and cell biology","confidence":"Medium","confidence_rationale":"Tier 2 — epistasis with multiple pathway interventions, single lab","pmids":["19767821"],"is_preprint":false},{"year":2012,"finding":"Caveolin-1 (Cav1) promotes WNT6 expression in gastric cancer cells; epirubicin increases WNT6 promoter activity through Cav1-dependent binding of β-catenin to the proximal WNT6 promoter; WNT6 increases resistance to epirubicin/doxorubicin-induced apoptosis.","method":"WNT6 knockdown and overexpression; WNT6 promoter reporter assay; β-catenin ChIP on WNT6 promoter; apoptosis assays; Cav1 manipulation","journal":"Oncogene","confidence":"High","confidence_rationale":"Tier 2 — ChIP demonstrating direct β-catenin binding to WNT6 promoter plus loss/gain-of-function functional assays","pmids":["22370641"],"is_preprint":false},{"year":2018,"finding":"PLAGL2 transcriptionally activates WNT6 by directly binding to its promoter region, thereby activating the Wnt/β-catenin pathway and promoting colorectal cancer cell proliferation.","method":"ChIP assay; dual-luciferase reporter assay; PLAGL2 knockdown; Wnt/β-catenin pathway gene expression analysis; xenograft model","journal":"Oncology reports","confidence":"High","confidence_rationale":"Tier 1–2 — ChIP plus reporter assay demonstrating direct transcription factor–promoter interaction with functional validation","pmids":["30535429"],"is_preprint":false},{"year":2019,"finding":"Secreted Wnt6 binds to receptor FZD4 to activate canonical β-catenin signaling, which is upstream of ROCK1 and 14-3-3σ, mediating diabetes-associated centrosome amplification; antibodies against Wnt6 or FZD4, or siRNA knockdown of either, attenuate centrosome amplification induced by high glucose/insulin/palmitic acid or advanced glycation end products (AGEs).","method":"siRNA knockdown of Wnt6, FZD4, β-catenin; neutralizing antibodies against Wnt6 and FZD4; β-catenin nuclear translocation assay; centrosome amplification quantification; in vivo diabetic mouse colon tissue analysis","journal":"American journal of physiology. Cell physiology","confidence":"High","confidence_rationale":"Tier 2 — receptor identification with multiple loss-of-function approaches (siRNA + antibody) and downstream pathway epistasis","pmids":["31618077"],"is_preprint":false},{"year":2020,"finding":"Wnt6 overexpression in the amygdala of MeCP2 T158A mice ameliorates behavioral deficits; Wnt6 restores GSK-3β phosphorylation and reduces β-catenin phosphorylation, restores MeCP2 SUMOylation, and increases BDNF and IGF-1 expression through activation of their promoters in a dose-dependent manner.","method":"Lentiviral Wnt6 delivery to amygdala; behavioral testing; Western blot for GSK-3β/β-catenin phosphorylation; SUMOylation assay; Bdnf/Igf-1 promoter reporter in HEK293T cells; RT-PCR and protein expression","journal":"Scientific reports","confidence":"Medium","confidence_rationale":"Tier 2 — in vivo lentiviral rescue with molecular pathway readouts, single lab","pmids":["31974426"],"is_preprint":false},{"year":2019,"finding":"HBCD (hexabromocyclododecane) promotes adipogenesis by specifically inhibiting Wnt6 gene expression and thereby suppressing the Wnt/β-catenin pathway at an early stage of adipogenesis; this was demonstrated both in cultured preadipocytes and in vivo in mice.","method":"RNA sequencing; Wnt6 expression analysis; adipogenesis assays in 3T3-L1 and HPA-V cells; in vivo epididymal fat analysis in C57BL/6 mice; Wnt/β-catenin pathway markers","journal":"The Science of the total environment","confidence":"Medium","confidence_rationale":"Tier 2 — RNA-seq plus in vitro/in vivo validation, mechanism via Wnt/β-catenin pathway, single lab","pmids":["31865202"],"is_preprint":false},{"year":2018,"finding":"In Drosophila ovaries, Wnt6 produced by cap cells is required to maintain anterior escort cells as a niche component; loss of Wnt6 signaling causes anterior escort cell death, leading to loss of germline stem cells; anterior escort cells maintain GSCs by promoting DE-cadherin anchorage and transient Dpp/BMP signaling.","method":"Drosophila genetic loss-of-function; hid expression-induced escort cell death; GSC counting; immunostaining for DE-cadherin, Dpp; live imaging","journal":"Development","confidence":"High","confidence_rationale":"Tier 2 — genetic loss-of-function with defined cellular and molecular mechanism in defined niche, Drosophila ortholog","pmids":["29361569"],"is_preprint":false},{"year":2021,"finding":"In Drosophila testis, Wnt6 regulates germline stem cell differentiation and cyst stem cell self-renewal via non-canonical Wnt signaling through Rac1 and Cdc42; Wnt6 downregulates Arm (β-catenin), Rac1, and Cdc42 in S2 cells, and Rac1/Cdc42 act downstream of Wnt6 in the niche.","method":"Drosophila genetic manipulation; Schneider 2 (S2) cell Wnt6 manipulation; Rac1, Cdc42, Arm protein level measurement; rescue epistasis experiments","journal":"Experimental cell research","confidence":"Medium","confidence_rationale":"Tier 2 — genetic epistasis in Drosophila with in vitro mechanistic validation, Drosophila ortholog","pmids":["33582096"],"is_preprint":false},{"year":2013,"finding":"Wnt6 is required for maxillary palp development in Drosophila; Wnt6 knockout flies lack maxillary palps but show no defects in wing size or planar cell polarity.","method":"CRISPR/homologous recombination Wnt6 knockout in Drosophila; morphological analysis of maxillary palps and wings","journal":"BMC biology","confidence":"High","confidence_rationale":"Tier 2 — clean genetic KO with specific organ-loss phenotype, Drosophila ortholog","pmids":["24090348"],"is_preprint":false},{"year":2011,"finding":"Maternal Wnt6 is necessary for endoderm specification in sea urchin embryos via canonical Wnt/β-catenin pathway activation; Wnt6 knockdown produces embryos lacking endoderm but retaining mesoderm markers; Wnt6 acts together with a localized vegetal cortical Dishevelled (Dsh) concentration to activate the endomesoderm gene regulatory network.","method":"Morpholino knockdown in sea urchin; vegetal cortex ablation and rescue with Wnt6; activated β-catenin/DN-GSK-3β/Dsh rescue; in situ hybridization for endoderm and mesoderm markers","journal":"Development","confidence":"Medium","confidence_rationale":"Tier 2 — morpholino KD with rescue experiments and pathway epistasis, invertebrate model","pmids":["21750039"],"is_preprint":false},{"year":2012,"finding":"Sulf1A is required for Wnt6-induced cellular hyper-elongation and inhibition of proliferation in myoblasts; Sulf1A neutralizing antibodies abolish both Wnt6-induced effects, demonstrating Sulf1A as a regulator of Wnt6 signaling in skeletal muscle.","method":"Wnt6 conditioned medium treatment of satellite cells on single fibres; Sulf1A neutralizing antibody treatment; proliferation and morphology assays","journal":"FEBS open bio","confidence":"Medium","confidence_rationale":"Tier 3 — single functional assay with antibody perturbation, single lab","pmids":["23772371"],"is_preprint":false},{"year":2022,"finding":"Wnt6 activates the β-catenin-dependent canonical pathway in human dental papilla cells, as shown by β-catenin nuclear accumulation, LEF1 upregulation, and enhanced TCF/Lef transcriptional activity; Wnt6 inhibits odontogenic differentiation in vitro but promotes mineralization after subcutaneous transplantation in vivo; XAV939 (Wnt/β-catenin inhibitor) attenuates Wnt6-induced mineralization.","method":"Wnt6 overexpression and knockdown; immunofluorescence for nuclear β-catenin; qPCR for LEF1; TOPflash/FOPflash dual-luciferase reporter; ALP staining; Alizarin Red; subcutaneous transplantation; XAV939 inhibitor treatment","journal":"Archives of oral biology","confidence":"Medium","confidence_rationale":"Tier 2 — multiple orthogonal assays confirming canonical pathway activation and functional consequence, single lab","pmids":["35691114"],"is_preprint":false},{"year":2017,"finding":"Wnt6 promotes viability and cell cycle progression (increasing S and G2/M phases) of mouse embryonic palatal mesenchymal cells through activation of the β-catenin signaling pathway; blockade of Wnt/β-catenin reduces Wnt6-induced cell viability.","method":"Recombinant Wnt6 treatment; MTT assay; flow cytometric cell cycle analysis; dual luciferase assay for β-catenin pathway; β-catenin pathway blockade","journal":"Experimental and therapeutic medicine","confidence":"Medium","confidence_rationale":"Tier 2 — pharmacological pathway blockade with functional readouts, single lab","pmids":["29285061"],"is_preprint":false},{"year":2021,"finding":"In mouse embryonic stem cells, WNT6 silencing partially impairs endothelial differentiation induced by BMP4, and this dysfunction is rescued by addition of recombinant WNT6, placing WNT6 downstream of BMP4 in the endothelial commitment pathway.","method":"WNT6 siRNA silencing in ES/PA6 co-culture; recombinant WNT6 rescue; microarray gene expression profiling; endothelial differentiation assays","journal":"Angiogenesis","confidence":"Medium","confidence_rationale":"Tier 2 — loss-of-function plus rescue in BMP4 pathway context, single lab","pmids":["34478025"],"is_preprint":false},{"year":2022,"finding":"Siglec6 overexpression in trophoblasts downregulates WNT6 secretion, impairing vascular endothelial cell functions (proliferation, migration, invasion, tube formation); recombinant human Wnt6 reverses Siglec6-conditioned medium-mediated HUVEC suppression by reactivating the Wnt/β-catenin signaling pathway.","method":"Siglec6 overexpression; transcriptome sequencing; qPCR and ELISA for Wnt6; HUVEC functional assays; recombinant Wnt6 rescue; Wnt/β-catenin pathway analysis","journal":"Archives of biochemistry and biophysics","confidence":"Medium","confidence_rationale":"Tier 2 — transcriptome identification plus functional rescue with recombinant protein, single lab","pmids":["36113626"],"is_preprint":false},{"year":2020,"finding":"Eif2s3y promotes spermatogonial stem cell (SSC) proliferation via activation of the Wnt6/β-catenin signaling pathway; transgenic Eif2s3y mice have increased active β-catenin and elevated Wnt6 expression, identifying Eif2s3y–Wnt6–β-catenin as a regulatory axis in spermatogenesis.","method":"Transgenic Eif2s3y mouse generation; SSC proliferation assay; active β-catenin and Wnt6 protein/mRNA expression analysis by Western blot and qPCR","journal":"Biochimica et biophysica acta. Molecular cell research","confidence":"Medium","confidence_rationale":"Tier 2 — transgenic gain-of-function with pathway readouts, single lab","pmids":["32621839"],"is_preprint":false}],"current_model":"WNT6 is a secreted cysteine-rich glycoprotein that primarily activates canonical β-catenin–dependent signaling (through receptors including FZD4 and FZD7) to regulate mesenchymal stem cell fate (inhibiting adipogenesis, promoting osteoblastogenesis), epithelialization, kidney tubulogenesis, uterine decidualization, and macrophage lipid metabolism/polarization, while also engaging non-canonical JNK and Rac1/Cdc42 pathways in specific contexts (neural crest induction, dental pulp cell migration, Drosophila niche maintenance); its expression is transcriptionally regulated by GATA6, FOXA2, PLAGL2, and Caveolin-1/β-catenin, and it can function both cell-autonomously and as a secreted paracrine ligand upstream of downstream effectors including cyclin B1, c-Myc (via ERK), ACC2, ROCK1, and cardiogenic transcription factors."},"narrative":{"teleology":[{"year":2001,"claim":"Establishing the molecular identity of WNT6 as a secreted glycoprotein with a signal peptide and WNT domain, clustered with WNT10A on chromosome 2q35, provided the foundation for functional studies.","evidence":"Molecular cloning and sequence analysis of human WNT6 cDNA","pmids":["11350055"],"confidence":"Medium","gaps":["No functional assays performed","Post-translational lipid modifications not characterized","Receptor specificity unknown"]},{"year":2002,"claim":"Demonstrating that Wnt6 induces kidney tubulogenesis and rescues Wnt4-mutant mesenchyme established WNT6 as a functional canonical Wnt ligand acting upstream of Wnt4 in nephrogenesis.","evidence":"Co-culture of Wnt6-expressing cells with embryonic kidney mesenchyme; Wnt4-mutant rescue; Xenopus secondary axis assay","pmids":["11948913"],"confidence":"High","gaps":["Receptor mediating Wnt6 signaling in kidney not identified","Endogenous requirement (loss-of-function in mammalian kidney) not tested"]},{"year":2005,"claim":"Identification of Wnt6 as an ectodermal signal promoting limb myogenesis via Pax3/Myf5 and simultaneously inhibiting chondrogenesis (via Sox9 repression) revealed WNT6 as a key ectodermal organizer of limb mesenchymal fate.","evidence":"Ectopic Wnt6 expression and ectoderm ablation in chick limb bud with in situ hybridization for myogenic and chondrogenic markers","pmids":["16271265","20334703"],"confidence":"High","gaps":["Canonical vs. non-canonical pathway usage in limb myogenesis not resolved","Direct Wnt6 receptor in limb mesenchyme unknown"]},{"year":2007,"claim":"Two parallel discoveries showed WNT6 activates canonical β-catenin–Snail1 signaling for primitive endoderm epithelialization in F9 cells, yet uses a non-canonical pathway for neural crest induction in chick, establishing context-dependent pathway selection.","evidence":"Wnt6 conditioned medium and overexpression in F9 cells with β-catenin/Snail1 nuclear translocation; in ovo Wnt6 gain/loss-of-function with canonical vs. non-canonical reporters in chick","pmids":["18160257","17685490"],"confidence":"High","gaps":["Molecular basis for pathway switching between canonical and non-canonical unknown","Non-canonical receptor and transducer in neural crest not identified"]},{"year":2008,"claim":"Wnt6 loss-of-function in Xenopus causing enlarged hearts, rescued by GATA, established WNT6 as a repressor of cardiomyogenesis via suppression of cardiogenic transcription factors.","evidence":"Xenopus morpholino knockdown; activated canonical Wnt gain-of-function; GATA rescue epistasis","pmids":["18804460"],"confidence":"High","gaps":["Mammalian heart phenotype of Wnt6 loss not reported","Direct vs. indirect repression of GATA factors unresolved"]},{"year":2011,"claim":"β-catenin epistasis experiments confirmed that WNT6 inhibits adipogenesis and promotes osteoblastogenesis through an obligate β-catenin-dependent mechanism in mesenchymal progenitors.","evidence":"Gain/loss-of-function in ST2 and 3T3-L1 cells; β-catenin knockdown abolishes effects","pmids":["21872687"],"confidence":"High","gaps":["Receptor identity for Wnt6 in mesenchymal stem cells not determined","In vivo bone/fat phenotype of Wnt6 knockout not examined"]},{"year":2012,"claim":"ChIP demonstrated that GATA6 and FOXA2 directly bind the WNT6 promoter, and separately that Caveolin-1/β-catenin binds the WNT6 promoter in gastric cancer, establishing the transcriptional regulation of WNT6 itself.","evidence":"ChIP and promoter reporter assays in F9 cells (GATA6/FOXA2) and gastric cancer cells (Cav1/β-catenin)","pmids":["22607194","22370641"],"confidence":"High","gaps":["Whether WNT6 autoregulation via β-catenin creates a positive feedback loop in normal tissue not tested","Chromatin accessibility and epigenetic regulation of WNT6 locus unexplored"]},{"year":2013,"claim":"Three advances defined WNT6 functions in uterine decidualization (via cyclin B1), macrophage biology (ERK–c-Myc proliferation and M2 polarization), and Drosophila appendage development, broadening WNT6's known tissue repertoire.","evidence":"Wnt6-mutant mouse uterine stromal cells; Wnt6 conditioned medium on macrophages with pertussis toxin/ERK inhibition; Drosophila Wnt6 CRISPR knockout","pmids":["23175771","24123681","24090348"],"confidence":"High","gaps":["Macrophage receptor for Wnt6 not identified","How Wnt6 activates pertussis toxin-sensitive Gαi in macrophages is mechanistically unclear"]},{"year":2015,"claim":"Sfrp2 was shown to bind Wnt6 and switch signaling from canonical to non-canonical JNK/PCP in cardiac progenitor cells, providing a molecular mechanism for the canonical/non-canonical pathway toggle.","evidence":"Sfrp2-Wnt6 binding assay; canonical Wnt reporter inhibition; JNK activation and CPC differentiation assays","pmids":["26071893"],"confidence":"High","gaps":["Structural basis of Sfrp2-Wnt6 interaction not determined","Whether other SFRPs similarly toggle Wnt6 signaling unknown"]},{"year":2016,"claim":"FZD7 was identified as a WNT6 receptor in renal epithelial cells, with Wnt6 signaling through GSK3β phosphorylation/β-catenin nuclear accumulation and additionally blocking TGF-β–NF-κB crosstalk.","evidence":"TCF/LEF reporter; GSK3β phosphorylation; β-catenin fractionation; p65 localization; p65−/− and IKKα/β−/− fibroblasts","pmids":["27122540"],"confidence":"High","gaps":["Whether FZD7 is the primary receptor in other WNT6-responsive tissues not established","Co-receptor (LRP5/6) requirement not formally tested"]},{"year":2019,"claim":"FZD4 was identified as a second WNT6 receptor, mediating canonical signaling upstream of ROCK1 and 14-3-3σ to drive centrosome amplification under diabetic conditions.","evidence":"siRNA and neutralizing antibodies against Wnt6/FZD4; β-catenin nuclear translocation; centrosome counting in diabetic mouse colon","pmids":["31618077"],"confidence":"High","gaps":["Whether FZD4 and FZD7 serve redundant or tissue-specific roles for WNT6 is unknown","Direct Wnt6–FZD4 physical binding not shown by biophysical methods"]},{"year":2021,"claim":"WNT6 was shown to promote macrophage foam cell formation via ACC2-dependent triacylglycerol storage, directly linking WNT6 to lipid metabolism and Mycobacterium tuberculosis intracellular survival.","evidence":"Wnt6-deficient macrophages; ACC2 pharmacological inhibition; intracellular TAG quantification; Mtb survival in vitro and in vivo","pmids":["34255743"],"confidence":"High","gaps":["How WNT6 regulates ACC2 expression or activity at the molecular level not defined","Whether this pathway operates in human TB macrophages in patients not confirmed"]},{"year":null,"claim":"Key unresolved questions include the structural basis of WNT6–Frizzled selectivity, the molecular switch determining canonical versus non-canonical pathway engagement, the phenotype of mammalian Wnt6 complete knockout across tissues, and whether WNT6 lipid modifications govern receptor specificity.","evidence":"","pmids":[],"confidence":"Low","gaps":["No crystal structure or cryo-EM of WNT6–receptor complex","No comprehensive mammalian Wnt6 knockout phenotyping across organ systems","Mechanism of canonical/non-canonical switch at the receptor level unresolved"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0048018","term_label":"receptor ligand activity","supporting_discovery_ids":[1,2,3,10,13,20,31]},{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[0,5,8,9,11]}],"localization":[{"term_id":"GO:0005576","term_label":"extracellular region","supporting_discovery_ids":[1,2,10,13,20,31]}],"pathway":[{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[0,3,4,10,12,13,15,20,24]},{"term_id":"R-HSA-1266738","term_label":"Developmental Biology","supporting_discovery_ids":[2,4,5,6,8,26,30]},{"term_id":"R-HSA-168256","term_label":"Immune System","supporting_discovery_ids":[10,11]},{"term_id":"R-HSA-1430728","term_label":"Metabolism","supporting_discovery_ids":[0,11,22]}],"complexes":[],"partners":["FZD4","FZD7","SFRP2","CTNNB1","ACC2","SULF1"],"other_free_text":[]},"mechanistic_narrative":"WNT6 is a secreted cysteine-rich glycoprotein that functions as a paracrine and autocrine ligand activating primarily canonical β-catenin–dependent signaling to regulate cell fate decisions across diverse developmental and adult tissue contexts. WNT6 signals through Frizzled receptors (FZD4, FZD7) to stabilize β-catenin, activate TCF/LEF transcription, and control mesenchymal stem cell fate by inhibiting adipogenesis and promoting osteoblastogenesis, while also driving kidney tubulogenesis (upstream of Wnt4), primitive endoderm epithelialization (via β-catenin–Snail1), uterine decidualization (via cyclin B1), and restriction of cardiomyogenesis [PMID:21872687, PMID:11948913, PMID:18160257, PMID:23175771, PMID:18804460]. In macrophages, WNT6 promotes proliferation through a β-catenin-independent ERK–c-Myc axis and drives M2 polarization and foam cell formation via ACC2-dependent lipid accumulation, facilitating intracellular Mycobacterium tuberculosis survival [PMID:24123681, PMID:34255743]. Context-dependent engagement of non-canonical pathways occurs in neural crest induction (chick ectoderm), dental pulp cell migration (JNK), and Drosophila niche maintenance (Rac1/Cdc42), and WNT6 transcription is directly regulated by GATA6, FOXA2, PLAGL2, and Caveolin-1/β-catenin [PMID:17685490, PMID:24935540, PMID:33582096, PMID:22607194, PMID:30535429, PMID:22370641]."},"prefetch_data":{"uniprot":{"accession":"Q9Y6F9","full_name":"Protein Wnt-6","aliases":[],"length_aa":365,"mass_kda":39.7,"function":"Ligand for members of the frizzled family of seven transmembrane receptors. Probable developmental protein. May be a signaling molecule which affects the development of discrete regions of tissues. Is likely to signal over only few cell diameters. 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epistasis; β-catenin stabilization assays\",\n      \"journal\": \"Bone\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — reciprocal gain/loss-of-function with epistasis (β-catenin KD), replicated across two cell lines\",\n      \"pmids\": [\"21872687\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2001,\n      \"finding\": \"Human WNT6 encodes a 365-amino-acid secreted glycoprotein with an N-terminal signal peptide, WNT core domain, and RGD motif; the WNT6 and WNT10A genes are clustered head-to-tail within 7 kb on chromosome 2q35.\",\n      \"method\": \"Molecular cloning, sequence analysis, Northern blot expression analysis\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — direct cloning and structural characterization, single lab\",\n      \"pmids\": [\"11350055\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2002,\n      \"finding\": \"Wnt-6 expressed in the ureter bud induces kidney tubulogenesis in vitro, rescues tubulogenesis in Wnt-4 mutant embryonic mesenchyme, and activates Wnt-4 transcription, placing Wnt-6 upstream of Wnt-4 in the tubulogenesis pathway; Wnt-6 also induces a secondary axis in Xenopus embryos, consistent with canonical pathway activation.\",\n      \"method\": \"Cell lines expressing Wnt-6 co-cultured with embryonic kidney mesenchyme; Wnt-4 mutant rescue assay; Xenopus secondary axis assay; in situ hybridization for tubulogenesis markers\",\n      \"journal\": \"Genesis\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — functional rescue in mutant background plus multiple orthogonal assays (tubulogenesis, marker induction, secondary axis)\",\n      \"pmids\": [\"11948913\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"Wnt6 induces specification and epithelialization of F9 embryonal carcinoma cells to primitive endoderm via a canonical Wnt6–β-catenin–Snail1 signaling cascade, involving nuclear translocation of β-catenin and Snail1 and appearance of cytokeratin intermediate filaments.\",\n      \"method\": \"Wnt6 conditioned medium treatment; Wnt6-expressing cell lines; LiCl GSK-3 inhibition; immunofluorescence for β-catenin and Snail1 nuclear translocation; cytokeratin staining; PKA activation for parietal endoderm EMT\",\n      \"journal\": \"Cellular signalling\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — multiple orthogonal assays (conditioned medium, overexpression, pharmacological) with defined molecular pathway\",\n      \"pmids\": [\"18160257\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"In chick embryos, Wnt6 derived from the surface ectoderm is necessary for neural crest induction and acts through the non-canonical (not β-catenin-dependent) signaling pathway; canonical Wnt signaling inhibited neural crest production in chick.\",\n      \"method\": \"In ovo Wnt6 loss-of-function and gain-of-function; canonical vs. non-canonical pathway reporter assays in chick embryos\",\n      \"journal\": \"Developmental dynamics\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — loss-of-function with specific neural crest phenotype and pathway discrimination by reporter assays\",\n      \"pmids\": [\"17685490\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"Wnt6 in Xenopus is required during organogenesis to restrict heart muscle development; Wnt6 loss-of-function results in abnormally large heart muscle, and this repressive role is mediated via repression of cardiogenic transcription factors (including GATA), since reinstating GATA function rescues cardiomyogenic differentiation.\",\n      \"method\": \"Xenopus loss-of-function (morpholinos); gain-of-function with activated canonical Wnt/β-catenin; GATA rescue epistasis; analysis of cardiogenic transcription factors and differentiation genes\",\n      \"journal\": \"Developmental biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — loss-of-function with defined phenotype, epistasis with GATA rescue, gain-of-function validation\",\n      \"pmids\": [\"18804460\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2005,\n      \"finding\": \"Ectodermal Wnt-6 promotes limb myogenesis through a Pax3/Paraxis–Myf5 pathway in chick; ectopic Wnt-6 expression extends domains of Pax3, Paraxis, Myf5, Myogenin, Desmin, and MyHC while inhibiting MyoD, and rescues myogenic gene expression after ectoderm removal, identifying a distinct Wnt-6 pathway separate from a MyoD-dependent ectodermal signal.\",\n      \"method\": \"Ectopic Wnt-6 expression in chick limb bud in ovo; ectoderm removal; in situ hybridization for myogenic markers\",\n      \"journal\": \"Developmental biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — gain-of-function and ectoderm ablation rescue with multiple marker readouts in vivo\",\n      \"pmids\": [\"16271265\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"Neural tube-derived Wnt1 and Wnt3a inhibit Wnt6 expression in the medial surface ectoderm via dermomyotomal Wnt11; Wnt11 was identified as an epithelialization factor acting on the medial dermomyotome, placing Wnt6 and Wnt11 as factors maintaining the epithelial nature of the ventrolateral and dorsomedial dermomyotomal lips, respectively.\",\n      \"method\": \"Interference with neighboring signaling pathways in chick embryo; in ovo gain/loss-of-function; in situ hybridization\",\n      \"journal\": \"Development\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — epistasis pathway placement in vivo, single lab\",\n      \"pmids\": [\"16818447\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"Ectodermal Wnt6 acts as a negative regulator of limb chondrogenesis in the chick embryo; ectoderm ablation leads to ectopic Sox9 and Collagen II expression, and Wnt6 overexpression reduces Sox9 and Collagen II, placing Wnt6 upstream of Sox9 in the chondrogenic cascade.\",\n      \"method\": \"Ectoderm ablation in chick limb bud; ectopic Wnt6 expression; in situ hybridization for Sox9, Collagen II\",\n      \"journal\": \"BMC developmental biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — reciprocal gain/loss-of-function with consistent upstream pathway placement, in vivo\",\n      \"pmids\": [\"20334703\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"Wnt6 is essential for uterine stromal cell proliferation during decidualization in mice; Wnt6 deficiency impairs stromal cell proliferation by downregulating cyclin B1 expression and prolonging cell cycle length, without adversely affecting decidual polyploidization.\",\n      \"method\": \"Wnt6-mutant mouse analysis; primary stromal cell culture; cell cycle analysis; cyclin B1 expression assay\",\n      \"journal\": \"Biology of reproduction\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — genetic KO with defined molecular mechanism (cyclin B1) and specific cellular phenotype\",\n      \"pmids\": [\"23175771\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"Wnt6 induces macrophage proliferation via a pertussis toxin-sensitive, ERK-mediated, β-catenin-independent induction of c-Myc; Wnt6 also drives macrophage polarization toward an M2-like phenotype by inducing Arginase-1 and downregulating TNF-α; Wnt6 expression in macrophages is induced via a TLR–MyD88–NF-κB-dependent pathway.\",\n      \"method\": \"Wnt6 conditioned medium treatment of macrophages; Wnt6-deficient macrophage analysis; thymidine incorporation proliferation assay; Ki-67 expression; pertussis toxin inhibition; ERK pathway analysis; Arginase-1 and TNF-α measurement\",\n      \"journal\": \"Journal of immunology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — loss-of-function, pharmacological inhibition (pertussis toxin), multiple orthogonal pathway readouts\",\n      \"pmids\": [\"24123681\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"WNT6 promotes foam cell formation in macrophages by regulating acetyl-CoA carboxylase 2 (ACC2) and triacylglycerol (TAG) storage; genetic or pharmacological loss of WNT6 or ACC2 significantly reduced intracellular TAG levels and Mycobacterium tuberculosis survival in macrophages.\",\n      \"method\": \"Genetic (Wnt6-deficient macrophages) and pharmacological (ACC2 inhibitor) approaches; intracellular TAG measurement; Mtb survival assay; in vivo mouse TB model\",\n      \"journal\": \"Journal of Clinical Investigation\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 — genetic KO plus pharmacological inhibition with defined lipid metabolic mechanism and in vivo validation\",\n      \"pmids\": [\"34255743\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"Sfrp2 binds Wnt6 and inhibits Wnt6 canonical signaling in cardiac progenitor cells (CPCs), which is required for inhibition of CPC proliferation; this canonical Wnt6 inhibition by Sfrp2 activates the non-canonical Wnt/PCP pathway through JNK, inducing cardiac transcription factors and CPC differentiation.\",\n      \"method\": \"Sfrp2-Wnt6 binding assay; canonical Wnt pathway reporter; JNK activation assay; CPC proliferation and differentiation assays; loss-of-function experiments\",\n      \"journal\": \"Journal of molecular and cellular cardiology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — direct binding demonstrated, pathway switching (canonical to non-canonical) with functional readouts\",\n      \"pmids\": [\"26071893\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"Wnt6 signals through the canonical pathway in renal epithelial cells via the FZD7 receptor, as evidenced by increased GSK3β phosphorylation (Ser9), nuclear accumulation of β-catenin, and increased TCF/Lef transcriptional activity; Wnt6 also inhibits TGF-β-mediated p65-NF-κB nuclear translocation and rescues epithelial dedifferentiation.\",\n      \"method\": \"TCF/Lef reporter assay; GSK3β phosphorylation Western blot; β-catenin nuclear fractionation; 3D tubulogenesis assay; TGF-β rescue experiments; p65-NF-κB localization; p65(-/-) and IKKα/β(-/-) fibroblast experiments\",\n      \"journal\": \"American journal of physiology. Renal physiology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — multiple orthogonal assays identifying receptor (FZD7), downstream signaling, and functional consequence\",\n      \"pmids\": [\"27122540\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"Wnt6 overexpression in human dental papilla cells enhances odontoblast-like differentiation (increased ALPase activity, upregulation of ALP, Col I, ON, OPN, BSP, DMP-1 gene expression, and mineral deposition) without significant effect on cell proliferation.\",\n      \"method\": \"Recombinant adenovirus Wnt6 overexpression; BrdU incorporation assay; flow cytometry; ALPase activity assay; mineralization assay; gene expression analysis\",\n      \"journal\": \"Journal of endodontics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — gain-of-function with multiple differentiation markers, single lab\",\n      \"pmids\": [\"20113781\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"WNT6 activates the JNK (c-Jun N-terminal kinase) non-canonical signaling pathway in human dental pulp cells, promoting cell migration and odontoblast-like differentiation; blockage of JNK with SP600125 decreases but does not completely abolish WNT6-induced migration and differentiation, indicating additional pathway involvement.\",\n      \"method\": \"WNT6 conditioned medium treatment; JNK inhibitor SP600125; Western blot for JNK activation; RT-PCR for c-Jun; wound healing and transwell migration assays; ALP and Alizarin red staining; odontogenic gene expression\",\n      \"journal\": \"Journal of endodontics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — pharmacological inhibition with multiple functional readouts, single lab\",\n      \"pmids\": [\"24935540\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"GATA6 and FOXA2 transcription factors directly bind the Wnt6 promoter and are sufficient to activate Wnt6 gene expression, leading to canonical WNT-β-catenin pathway activation and primitive endoderm induction in F9 cells.\",\n      \"method\": \"Chromatin immunoprecipitation (ChIP); Wnt6 promoter reporter assay; GATA6/FOXA2 overexpression; TCF-dependent transcription assay\",\n      \"journal\": \"Stem cells and development\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 — ChIP demonstrating direct promoter binding plus reporter and overexpression validation\",\n      \"pmids\": [\"22607194\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"Coordinate Gα13–RhoA and Wnt6–β-catenin signaling are both required for retinoic acid-induced differentiation of F9 cells to primitive endoderm; Gα13/p115RhoGEF (but not RhoA alone) causes β-catenin nuclear translocation, and two independent pathways (canonical Wnt-β-catenin and Gα13-ERM) must both be active for primitive endoderm formation.\",\n      \"method\": \"Constitutively active Gα13(Q226L), p115RhoGEF, and RhoA(L63) overexpression; dominant-negative constructs; ROCK inhibitor Y-27632; β-catenin nuclear localization assay; F9 differentiation markers\",\n      \"journal\": \"Biochemistry and cell biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — epistasis with multiple pathway interventions, single lab\",\n      \"pmids\": [\"19767821\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"Caveolin-1 (Cav1) promotes WNT6 expression in gastric cancer cells; epirubicin increases WNT6 promoter activity through Cav1-dependent binding of β-catenin to the proximal WNT6 promoter; WNT6 increases resistance to epirubicin/doxorubicin-induced apoptosis.\",\n      \"method\": \"WNT6 knockdown and overexpression; WNT6 promoter reporter assay; β-catenin ChIP on WNT6 promoter; apoptosis assays; Cav1 manipulation\",\n      \"journal\": \"Oncogene\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — ChIP demonstrating direct β-catenin binding to WNT6 promoter plus loss/gain-of-function functional assays\",\n      \"pmids\": [\"22370641\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"PLAGL2 transcriptionally activates WNT6 by directly binding to its promoter region, thereby activating the Wnt/β-catenin pathway and promoting colorectal cancer cell proliferation.\",\n      \"method\": \"ChIP assay; dual-luciferase reporter assay; PLAGL2 knockdown; Wnt/β-catenin pathway gene expression analysis; xenograft model\",\n      \"journal\": \"Oncology reports\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 — ChIP plus reporter assay demonstrating direct transcription factor–promoter interaction with functional validation\",\n      \"pmids\": [\"30535429\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"Secreted Wnt6 binds to receptor FZD4 to activate canonical β-catenin signaling, which is upstream of ROCK1 and 14-3-3σ, mediating diabetes-associated centrosome amplification; antibodies against Wnt6 or FZD4, or siRNA knockdown of either, attenuate centrosome amplification induced by high glucose/insulin/palmitic acid or advanced glycation end products (AGEs).\",\n      \"method\": \"siRNA knockdown of Wnt6, FZD4, β-catenin; neutralizing antibodies against Wnt6 and FZD4; β-catenin nuclear translocation assay; centrosome amplification quantification; in vivo diabetic mouse colon tissue analysis\",\n      \"journal\": \"American journal of physiology. Cell physiology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — receptor identification with multiple loss-of-function approaches (siRNA + antibody) and downstream pathway epistasis\",\n      \"pmids\": [\"31618077\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"Wnt6 overexpression in the amygdala of MeCP2 T158A mice ameliorates behavioral deficits; Wnt6 restores GSK-3β phosphorylation and reduces β-catenin phosphorylation, restores MeCP2 SUMOylation, and increases BDNF and IGF-1 expression through activation of their promoters in a dose-dependent manner.\",\n      \"method\": \"Lentiviral Wnt6 delivery to amygdala; behavioral testing; Western blot for GSK-3β/β-catenin phosphorylation; SUMOylation assay; Bdnf/Igf-1 promoter reporter in HEK293T cells; RT-PCR and protein expression\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — in vivo lentiviral rescue with molecular pathway readouts, single lab\",\n      \"pmids\": [\"31974426\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"HBCD (hexabromocyclododecane) promotes adipogenesis by specifically inhibiting Wnt6 gene expression and thereby suppressing the Wnt/β-catenin pathway at an early stage of adipogenesis; this was demonstrated both in cultured preadipocytes and in vivo in mice.\",\n      \"method\": \"RNA sequencing; Wnt6 expression analysis; adipogenesis assays in 3T3-L1 and HPA-V cells; in vivo epididymal fat analysis in C57BL/6 mice; Wnt/β-catenin pathway markers\",\n      \"journal\": \"The Science of the total environment\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — RNA-seq plus in vitro/in vivo validation, mechanism via Wnt/β-catenin pathway, single lab\",\n      \"pmids\": [\"31865202\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"In Drosophila ovaries, Wnt6 produced by cap cells is required to maintain anterior escort cells as a niche component; loss of Wnt6 signaling causes anterior escort cell death, leading to loss of germline stem cells; anterior escort cells maintain GSCs by promoting DE-cadherin anchorage and transient Dpp/BMP signaling.\",\n      \"method\": \"Drosophila genetic loss-of-function; hid expression-induced escort cell death; GSC counting; immunostaining for DE-cadherin, Dpp; live imaging\",\n      \"journal\": \"Development\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — genetic loss-of-function with defined cellular and molecular mechanism in defined niche, Drosophila ortholog\",\n      \"pmids\": [\"29361569\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"In Drosophila testis, Wnt6 regulates germline stem cell differentiation and cyst stem cell self-renewal via non-canonical Wnt signaling through Rac1 and Cdc42; Wnt6 downregulates Arm (β-catenin), Rac1, and Cdc42 in S2 cells, and Rac1/Cdc42 act downstream of Wnt6 in the niche.\",\n      \"method\": \"Drosophila genetic manipulation; Schneider 2 (S2) cell Wnt6 manipulation; Rac1, Cdc42, Arm protein level measurement; rescue epistasis experiments\",\n      \"journal\": \"Experimental cell research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — genetic epistasis in Drosophila with in vitro mechanistic validation, Drosophila ortholog\",\n      \"pmids\": [\"33582096\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"Wnt6 is required for maxillary palp development in Drosophila; Wnt6 knockout flies lack maxillary palps but show no defects in wing size or planar cell polarity.\",\n      \"method\": \"CRISPR/homologous recombination Wnt6 knockout in Drosophila; morphological analysis of maxillary palps and wings\",\n      \"journal\": \"BMC biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — clean genetic KO with specific organ-loss phenotype, Drosophila ortholog\",\n      \"pmids\": [\"24090348\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"Maternal Wnt6 is necessary for endoderm specification in sea urchin embryos via canonical Wnt/β-catenin pathway activation; Wnt6 knockdown produces embryos lacking endoderm but retaining mesoderm markers; Wnt6 acts together with a localized vegetal cortical Dishevelled (Dsh) concentration to activate the endomesoderm gene regulatory network.\",\n      \"method\": \"Morpholino knockdown in sea urchin; vegetal cortex ablation and rescue with Wnt6; activated β-catenin/DN-GSK-3β/Dsh rescue; in situ hybridization for endoderm and mesoderm markers\",\n      \"journal\": \"Development\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — morpholino KD with rescue experiments and pathway epistasis, invertebrate model\",\n      \"pmids\": [\"21750039\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"Sulf1A is required for Wnt6-induced cellular hyper-elongation and inhibition of proliferation in myoblasts; Sulf1A neutralizing antibodies abolish both Wnt6-induced effects, demonstrating Sulf1A as a regulator of Wnt6 signaling in skeletal muscle.\",\n      \"method\": \"Wnt6 conditioned medium treatment of satellite cells on single fibres; Sulf1A neutralizing antibody treatment; proliferation and morphology assays\",\n      \"journal\": \"FEBS open bio\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 — single functional assay with antibody perturbation, single lab\",\n      \"pmids\": [\"23772371\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"Wnt6 activates the β-catenin-dependent canonical pathway in human dental papilla cells, as shown by β-catenin nuclear accumulation, LEF1 upregulation, and enhanced TCF/Lef transcriptional activity; Wnt6 inhibits odontogenic differentiation in vitro but promotes mineralization after subcutaneous transplantation in vivo; XAV939 (Wnt/β-catenin inhibitor) attenuates Wnt6-induced mineralization.\",\n      \"method\": \"Wnt6 overexpression and knockdown; immunofluorescence for nuclear β-catenin; qPCR for LEF1; TOPflash/FOPflash dual-luciferase reporter; ALP staining; Alizarin Red; subcutaneous transplantation; XAV939 inhibitor treatment\",\n      \"journal\": \"Archives of oral biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — multiple orthogonal assays confirming canonical pathway activation and functional consequence, single lab\",\n      \"pmids\": [\"35691114\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"Wnt6 promotes viability and cell cycle progression (increasing S and G2/M phases) of mouse embryonic palatal mesenchymal cells through activation of the β-catenin signaling pathway; blockade of Wnt/β-catenin reduces Wnt6-induced cell viability.\",\n      \"method\": \"Recombinant Wnt6 treatment; MTT assay; flow cytometric cell cycle analysis; dual luciferase assay for β-catenin pathway; β-catenin pathway blockade\",\n      \"journal\": \"Experimental and therapeutic medicine\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — pharmacological pathway blockade with functional readouts, single lab\",\n      \"pmids\": [\"29285061\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"In mouse embryonic stem cells, WNT6 silencing partially impairs endothelial differentiation induced by BMP4, and this dysfunction is rescued by addition of recombinant WNT6, placing WNT6 downstream of BMP4 in the endothelial commitment pathway.\",\n      \"method\": \"WNT6 siRNA silencing in ES/PA6 co-culture; recombinant WNT6 rescue; microarray gene expression profiling; endothelial differentiation assays\",\n      \"journal\": \"Angiogenesis\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — loss-of-function plus rescue in BMP4 pathway context, single lab\",\n      \"pmids\": [\"34478025\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"Siglec6 overexpression in trophoblasts downregulates WNT6 secretion, impairing vascular endothelial cell functions (proliferation, migration, invasion, tube formation); recombinant human Wnt6 reverses Siglec6-conditioned medium-mediated HUVEC suppression by reactivating the Wnt/β-catenin signaling pathway.\",\n      \"method\": \"Siglec6 overexpression; transcriptome sequencing; qPCR and ELISA for Wnt6; HUVEC functional assays; recombinant Wnt6 rescue; Wnt/β-catenin pathway analysis\",\n      \"journal\": \"Archives of biochemistry and biophysics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — transcriptome identification plus functional rescue with recombinant protein, single lab\",\n      \"pmids\": [\"36113626\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"Eif2s3y promotes spermatogonial stem cell (SSC) proliferation via activation of the Wnt6/β-catenin signaling pathway; transgenic Eif2s3y mice have increased active β-catenin and elevated Wnt6 expression, identifying Eif2s3y–Wnt6–β-catenin as a regulatory axis in spermatogenesis.\",\n      \"method\": \"Transgenic Eif2s3y mouse generation; SSC proliferation assay; active β-catenin and Wnt6 protein/mRNA expression analysis by Western blot and qPCR\",\n      \"journal\": \"Biochimica et biophysica acta. Molecular cell research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — transgenic gain-of-function with pathway readouts, single lab\",\n      \"pmids\": [\"32621839\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"WNT6 is a secreted cysteine-rich glycoprotein that primarily activates canonical β-catenin–dependent signaling (through receptors including FZD4 and FZD7) to regulate mesenchymal stem cell fate (inhibiting adipogenesis, promoting osteoblastogenesis), epithelialization, kidney tubulogenesis, uterine decidualization, and macrophage lipid metabolism/polarization, while also engaging non-canonical JNK and Rac1/Cdc42 pathways in specific contexts (neural crest induction, dental pulp cell migration, Drosophila niche maintenance); its expression is transcriptionally regulated by GATA6, FOXA2, PLAGL2, and Caveolin-1/β-catenin, and it can function both cell-autonomously and as a secreted paracrine ligand upstream of downstream effectors including cyclin B1, c-Myc (via ERK), ACC2, ROCK1, and cardiogenic transcription factors.\"\n}\n```","stage2_raw":"```json\n{\n  \"mechanistic_narrative\": \"WNT6 is a secreted cysteine-rich glycoprotein that functions as a paracrine and autocrine ligand activating primarily canonical β-catenin–dependent signaling to regulate cell fate decisions across diverse developmental and adult tissue contexts. WNT6 signals through Frizzled receptors (FZD4, FZD7) to stabilize β-catenin, activate TCF/LEF transcription, and control mesenchymal stem cell fate by inhibiting adipogenesis and promoting osteoblastogenesis, while also driving kidney tubulogenesis (upstream of Wnt4), primitive endoderm epithelialization (via β-catenin–Snail1), uterine decidualization (via cyclin B1), and restriction of cardiomyogenesis [PMID:21872687, PMID:11948913, PMID:18160257, PMID:23175771, PMID:18804460]. In macrophages, WNT6 promotes proliferation through a β-catenin-independent ERK–c-Myc axis and drives M2 polarization and foam cell formation via ACC2-dependent lipid accumulation, facilitating intracellular Mycobacterium tuberculosis survival [PMID:24123681, PMID:34255743]. Context-dependent engagement of non-canonical pathways occurs in neural crest induction (chick ectoderm), dental pulp cell migration (JNK), and Drosophila niche maintenance (Rac1/Cdc42), and WNT6 transcription is directly regulated by GATA6, FOXA2, PLAGL2, and Caveolin-1/β-catenin [PMID:17685490, PMID:24935540, PMID:33582096, PMID:22607194, PMID:30535429, PMID:22370641].\",\n  \"teleology\": [\n    {\n      \"year\": 2001,\n      \"claim\": \"Establishing the molecular identity of WNT6 as a secreted glycoprotein with a signal peptide and WNT domain, clustered with WNT10A on chromosome 2q35, provided the foundation for functional studies.\",\n      \"evidence\": \"Molecular cloning and sequence analysis of human WNT6 cDNA\",\n      \"pmids\": [\"11350055\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No functional assays performed\", \"Post-translational lipid modifications not characterized\", \"Receptor specificity unknown\"]\n    },\n    {\n      \"year\": 2002,\n      \"claim\": \"Demonstrating that Wnt6 induces kidney tubulogenesis and rescues Wnt4-mutant mesenchyme established WNT6 as a functional canonical Wnt ligand acting upstream of Wnt4 in nephrogenesis.\",\n      \"evidence\": \"Co-culture of Wnt6-expressing cells with embryonic kidney mesenchyme; Wnt4-mutant rescue; Xenopus secondary axis assay\",\n      \"pmids\": [\"11948913\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Receptor mediating Wnt6 signaling in kidney not identified\", \"Endogenous requirement (loss-of-function in mammalian kidney) not tested\"]\n    },\n    {\n      \"year\": 2005,\n      \"claim\": \"Identification of Wnt6 as an ectodermal signal promoting limb myogenesis via Pax3/Myf5 and simultaneously inhibiting chondrogenesis (via Sox9 repression) revealed WNT6 as a key ectodermal organizer of limb mesenchymal fate.\",\n      \"evidence\": \"Ectopic Wnt6 expression and ectoderm ablation in chick limb bud with in situ hybridization for myogenic and chondrogenic markers\",\n      \"pmids\": [\"16271265\", \"20334703\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Canonical vs. non-canonical pathway usage in limb myogenesis not resolved\", \"Direct Wnt6 receptor in limb mesenchyme unknown\"]\n    },\n    {\n      \"year\": 2007,\n      \"claim\": \"Two parallel discoveries showed WNT6 activates canonical β-catenin–Snail1 signaling for primitive endoderm epithelialization in F9 cells, yet uses a non-canonical pathway for neural crest induction in chick, establishing context-dependent pathway selection.\",\n      \"evidence\": \"Wnt6 conditioned medium and overexpression in F9 cells with β-catenin/Snail1 nuclear translocation; in ovo Wnt6 gain/loss-of-function with canonical vs. non-canonical reporters in chick\",\n      \"pmids\": [\"18160257\", \"17685490\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Molecular basis for pathway switching between canonical and non-canonical unknown\", \"Non-canonical receptor and transducer in neural crest not identified\"]\n    },\n    {\n      \"year\": 2008,\n      \"claim\": \"Wnt6 loss-of-function in Xenopus causing enlarged hearts, rescued by GATA, established WNT6 as a repressor of cardiomyogenesis via suppression of cardiogenic transcription factors.\",\n      \"evidence\": \"Xenopus morpholino knockdown; activated canonical Wnt gain-of-function; GATA rescue epistasis\",\n      \"pmids\": [\"18804460\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Mammalian heart phenotype of Wnt6 loss not reported\", \"Direct vs. indirect repression of GATA factors unresolved\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"β-catenin epistasis experiments confirmed that WNT6 inhibits adipogenesis and promotes osteoblastogenesis through an obligate β-catenin-dependent mechanism in mesenchymal progenitors.\",\n      \"evidence\": \"Gain/loss-of-function in ST2 and 3T3-L1 cells; β-catenin knockdown abolishes effects\",\n      \"pmids\": [\"21872687\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Receptor identity for Wnt6 in mesenchymal stem cells not determined\", \"In vivo bone/fat phenotype of Wnt6 knockout not examined\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"ChIP demonstrated that GATA6 and FOXA2 directly bind the WNT6 promoter, and separately that Caveolin-1/β-catenin binds the WNT6 promoter in gastric cancer, establishing the transcriptional regulation of WNT6 itself.\",\n      \"evidence\": \"ChIP and promoter reporter assays in F9 cells (GATA6/FOXA2) and gastric cancer cells (Cav1/β-catenin)\",\n      \"pmids\": [\"22607194\", \"22370641\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether WNT6 autoregulation via β-catenin creates a positive feedback loop in normal tissue not tested\", \"Chromatin accessibility and epigenetic regulation of WNT6 locus unexplored\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Three advances defined WNT6 functions in uterine decidualization (via cyclin B1), macrophage biology (ERK–c-Myc proliferation and M2 polarization), and Drosophila appendage development, broadening WNT6's known tissue repertoire.\",\n      \"evidence\": \"Wnt6-mutant mouse uterine stromal cells; Wnt6 conditioned medium on macrophages with pertussis toxin/ERK inhibition; Drosophila Wnt6 CRISPR knockout\",\n      \"pmids\": [\"23175771\", \"24123681\", \"24090348\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Macrophage receptor for Wnt6 not identified\", \"How Wnt6 activates pertussis toxin-sensitive Gαi in macrophages is mechanistically unclear\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Sfrp2 was shown to bind Wnt6 and switch signaling from canonical to non-canonical JNK/PCP in cardiac progenitor cells, providing a molecular mechanism for the canonical/non-canonical pathway toggle.\",\n      \"evidence\": \"Sfrp2-Wnt6 binding assay; canonical Wnt reporter inhibition; JNK activation and CPC differentiation assays\",\n      \"pmids\": [\"26071893\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Structural basis of Sfrp2-Wnt6 interaction not determined\", \"Whether other SFRPs similarly toggle Wnt6 signaling unknown\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"FZD7 was identified as a WNT6 receptor in renal epithelial cells, with Wnt6 signaling through GSK3β phosphorylation/β-catenin nuclear accumulation and additionally blocking TGF-β–NF-κB crosstalk.\",\n      \"evidence\": \"TCF/LEF reporter; GSK3β phosphorylation; β-catenin fractionation; p65 localization; p65−/− and IKKα/β−/− fibroblasts\",\n      \"pmids\": [\"27122540\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether FZD7 is the primary receptor in other WNT6-responsive tissues not established\", \"Co-receptor (LRP5/6) requirement not formally tested\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"FZD4 was identified as a second WNT6 receptor, mediating canonical signaling upstream of ROCK1 and 14-3-3σ to drive centrosome amplification under diabetic conditions.\",\n      \"evidence\": \"siRNA and neutralizing antibodies against Wnt6/FZD4; β-catenin nuclear translocation; centrosome counting in diabetic mouse colon\",\n      \"pmids\": [\"31618077\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether FZD4 and FZD7 serve redundant or tissue-specific roles for WNT6 is unknown\", \"Direct Wnt6–FZD4 physical binding not shown by biophysical methods\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"WNT6 was shown to promote macrophage foam cell formation via ACC2-dependent triacylglycerol storage, directly linking WNT6 to lipid metabolism and Mycobacterium tuberculosis intracellular survival.\",\n      \"evidence\": \"Wnt6-deficient macrophages; ACC2 pharmacological inhibition; intracellular TAG quantification; Mtb survival in vitro and in vivo\",\n      \"pmids\": [\"34255743\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How WNT6 regulates ACC2 expression or activity at the molecular level not defined\", \"Whether this pathway operates in human TB macrophages in patients not confirmed\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"Key unresolved questions include the structural basis of WNT6–Frizzled selectivity, the molecular switch determining canonical versus non-canonical pathway engagement, the phenotype of mammalian Wnt6 complete knockout across tissues, and whether WNT6 lipid modifications govern receptor specificity.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"No crystal structure or cryo-EM of WNT6–receptor complex\", \"No comprehensive mammalian Wnt6 knockout phenotyping across organ systems\", \"Mechanism of canonical/non-canonical switch at the receptor level unresolved\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0048018\", \"supporting_discovery_ids\": [1, 2, 3, 10, 13, 20, 31]},\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [0, 5, 8, 9, 11]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005576\", \"supporting_discovery_ids\": [1, 2, 10, 13, 20, 31]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"GO:0016055\", \"supporting_discovery_ids\": [0, 2, 3, 13, 20]},\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [0, 3, 4, 10, 12, 13, 15, 20, 24]},\n      {\"term_id\": \"R-HSA-1266738\", \"supporting_discovery_ids\": [2, 4, 5, 6, 8, 26, 30]},\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [10, 11]},\n      {\"term_id\": \"R-HSA-1430728\", \"supporting_discovery_ids\": [0, 11, 22]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\n      \"FZD4\",\n      \"FZD7\",\n      \"SFRP2\",\n      \"CTNNB1\",\n      \"ACC2\",\n      \"SULF1\"\n    ],\n    \"other_free_text\": []\n  }\n}\n```"}