| 2006 |
Purified Wnt5a protein inhibits Wnt3a-induced canonical Wnt signaling by downregulating beta-catenin-induced reporter gene expression (not by influencing beta-catenin levels) via the orphan tyrosine kinase Ror2. In the presence of Frizzled 4, the same purified Wnt5a activates beta-catenin signaling, demonstrating that receptor context dictates signaling output. The Wnt5a signal is pertussis toxin insensitive and does not influence cellular calcium levels in this context. |
Purified Wnt5a protein (homogeneous), dose-response reporter assays, Ror2 requirement established by functional blocking, Frizzled 4 co-expression experiment |
PLoS biology |
High |
16602827
|
| 2002 |
Wnt5a overexpression in melanoma cells causes actin reorganization, increased cell adhesion, and increased PKC activation without increasing beta-catenin expression or nuclear translocation. Blocking Frizzled-5 with antibodies inhibits PKC activity and cellular invasion, placing Frizzled-5 upstream of PKC in Wnt5a-driven melanoma invasion. |
Constitutive overexpression vector transfection, anti-Frizzled-5 antibody blocking, PKC activity assay, invasion assay, actin staining |
Cancer cell |
Medium |
12086864
|
| 2008 |
Wnt5a induces homodimerization and tyrosine phosphorylation of Ror2 in U2OS osteoblastic cells. Wnt5a also causes phosphorylation of the Ror2 substrate 14-3-3beta. Wnt3a binds Ror2 but does not induce its homodimerization or activation. Ror2 knockdown abolishes Wnt5a-induced osteoblastic differentiation of human mesenchymal stem cells. |
Co-immunoprecipitation, tyrosine phosphorylation assay, 14-3-3beta phosphorylation assay, siRNA knockdown of Ror2, calvarial bone explant culture, MSC differentiation assay |
Journal of cellular biochemistry |
Medium |
18615587
|
| 2007 |
Wnt5a induces serine/threonine phosphorylation of Ror2 through GSK-3 (primarily GSK-3alpha). GSK-3 inhibitors (LiCl, SB216763) or siRNA against GSK-3 block Wnt5a-induced Ror2 phosphorylation. Purified GSK-3alpha and GSK-3beta phosphorylate immunoprecipitated Ror2 in vitro. Wnt5a-induced cell migration is inhibited by GSK-3 suppression. |
In vitro kinase assay with purified GSK-3 on immunoprecipitated Ror2, siRNA knockdown of GSK-3, pharmacological inhibitors (LiCl, SB216763), cell migration assay |
Genes to cells |
High |
17986005
|
| 2008 |
Wnt5a controls cell polarity and directional movement in melanoma by recruiting actin, myosin IIB, Frizzled 3, and MCAM into an intracellular Wnt-mediated receptor-actin-myosin polarity (W-RAMP) structure. In a chemokine gradient, W-RAMP accumulates asymmetrically at the cell periphery, triggering membrane contractility and nuclear movement. The process requires endosome trafficking, is associated with multivesicular bodies, and is regulated through small GTPases Rab4 and RhoB. |
Live imaging, immunofluorescence, dominant-negative Rab4/RhoB expression, chemokine gradient assay, subcellular fractionation/endosome trafficking analysis |
Science |
High |
18420933
|
| 2009 |
ROR2 is required for Wnt5a-mediated melanoma metastasis. Wnt5a increases ROR2 expression and promotes PKC-dependent, clathrin-mediated internalization of ROR2. siRNA knockdown of ROR2 decreases downstream Wnt5a signaling and reduces melanoma metastasis in vitro and in vivo. |
siRNA knockdown of ROR2, in vitro and in vivo metastasis assays, PKC inhibitor, clathrin inhibitor, melanoma tissue microarrays |
Oncogene |
Medium |
19802008
|
| 2010 |
Wnt5a promotes prostate cancer cell invasion through Frizzled2 and Ror2 receptors. Wnt5a activates JNK through protein kinase D (PKD), and PKD inhibition suppresses Wnt5a-dependent cell migration and invasion. Wnt5a induces MMP-1 expression through recruitment of JunD to the MMP-1 promoter. |
Wnt5a knockdown and overexpression in prostate cancer cell lines, invasion assay, PKD inhibitor, ChIP assay for JunD at MMP-1 promoter, siRNA for Frizzled2/Ror2 |
Oncogene |
Medium |
20101234
|
| 2013 |
CD146 acts as a functional Wnt5a receptor: CD146 binds Wnt5a with high affinity, and Wnt5a treatment enhances CD146-Dvl2 interaction. CD146-mediated Wnt5a signaling activates Dvl and JNK to promote cell migration and zebrafish convergent extension. Mutation of the Dvl2-binding region of CD146 impairs JNK activation, cell migration, and cell protrusion formation. CD146 also inhibits canonical Wnt signaling by promoting beta-catenin degradation. |
Binding affinity assay, co-immunoprecipitation (CD146-Dvl2), domain mutation, JNK activation assay, cell migration assay, zebrafish knockdown/convergent extension assay, beta-catenin degradation assay |
Nature communications |
High |
24335906
|
| 2016 |
Wnt5a induces ROR1/ROR2 heterooligomerization to recruit guanine exchange factors (GEFs), activating Rac1 and RhoA in CLL cells. The extracellular Kringle domain of ROR1 is required for ROR1/ROR2 heterooligomerization; the cysteine-rich domain or intracellular proline-rich domain is required for Wnt5a-induced GEF recruitment. Silencing either ROR1 or ROR2 blocks these effects. |
Co-immunoprecipitation (ROR1/ROR2 heterooligomers), siRNA silencing of ROR1/ROR2, domain mapping with truncation constructs, Rac1/RhoA activation assays, ectopic ROR1 expression in ROR1-deficient MEC1 cells, in vivo engraftment assays in immune-deficient and ROR1-transgenic mice |
The Journal of clinical investigation |
High |
26690702
|
| 2015 |
Wnt5a signals through DVL1 to repress ribosomal DNA (rDNA) transcription by RNA Polymerase I. DVL1 accumulates at nucleolar organizer regions and binds rDNA regions. Upon DVL1 binding, SIRT7 (a Pol I transcription activator/deacetylase) dissociates from rDNA loci, concomitant with disassembly of the Pol I transcription machinery at the rDNA promoter. |
ChIP of DVL1 at rDNA, co-immunoprecipitation, Pol I transcription assay, SIRT7 displacement assay, DVL1-specific requirement established by siRNA |
PLoS genetics |
Medium |
27500936
|
| 2015 |
Wnt5a promotes protein depalmitoylation of MCAM at Cys-590 via acyl protein thioesterase 1 (APT1), resulting in asymmetric MCAM localization. Mutation of Cys-590 to glycine is sufficient to polarize MCAM localization. Inhibition of APT1 blocks Wnt5a-induced depalmitoylation, asymmetric MCAM localization, and cell invasion. |
Site-directed mutagenesis (Cys-590), APT1 inhibitor treatment, palmitoylation assay, immunofluorescence for MCAM localization, invasion assay |
The Journal of biological chemistry |
Medium |
25944911
|
| 2018 |
Wnt5a signaling induces APT1 phosphorylation, which increases APT1 depalmitoylating activity and reduces APT1 dimerization. Phosphorylated APT1 promotes depalmitoylation of pro-metastatic cell adhesion molecules CD44 and MCAM, increasing melanoma invasion in vitro. |
Phosphorylation assay of APT1, depalmitoylation activity assay, dimerization assay, invasion assay, correlation with tumor grade |
eLife |
Medium |
29648538
|
| 2014 |
Wnt5a-induced noncanonical signaling enhances Wnt/beta-catenin signaling during osteoblastogenesis by upregulating expression of co-receptors Lrp5 and Lrp6. Wnt5a knockdown reduces Lrp5/6 expression and impairs osteoblast differentiation while enhancing adipocyte differentiation. Adenovirus-mediated gene transfer of Lrp5 into Wnt5a-deficient osteoblast cells rescues the differentiation phenotype. |
Conditional Wnt5a knockout (osteoblast-lineage), shRNA knockdown, adenoviral Lrp5 rescue, Tcf/Lef reporter assay, differentiation assays |
Scientific reports |
Medium |
24670389
|
| 2016 |
Wnt5a regulates AQP2 water channel expression, phosphorylation, and apical membrane trafficking in renal collecting duct cells via calcineurin signaling, independently of the vasopressin/cAMP/PKA pathway. Wnt5a increases apical membrane AQP2 localization and urine osmolality in an NDI mouse model. Calcineurin activator arachidonic acid mimics Wnt5a effects. |
In vitro Wnt5a stimulation of collecting duct cells, AQP2 phosphorylation and trafficking assays, calcineurin activity assay, cAMP/PKA measurement, NDI mouse model in vivo |
Nature communications |
High |
27892464
|
| 2017 |
Wnt5a-Ror2 signaling in osteoclasts activates RhoA, which engages the Rho effector kinase Pkn3. Pkn3 binds to and enhances the activity of c-Src, promoting actin ring formation and bone resorption. Osteoclast-specific Ror2 knockout mice have increased bone mass and impaired actin ring formation and bone resorption, defects rescued by constitutively active RhoA. Pkn3 global knockout mice show similar increased bone mass phenotype. |
Osteoclast-specific Ror2 conditional knockout mice, Pkn3 global knockout mice, constitutively active RhoA rescue experiment, c-Src kinase activity assay, co-immunoprecipitation (Pkn3-c-Src), actin ring formation assay, bone resorption assay |
Science signaling |
High |
28851822
|
| 2016 |
Wnt5a signals through the Ryk receptor to regulate hematopoietic stem cell (HSC) quiescence and hematopoietic repopulation. Ryk inhibition blocks Wnt5a-induced HSC quiescence and suppression of reactive oxygen species (ROS) production, which is a known driver of HSC proliferation. |
Ryk blocking antibody or dominant-negative Ryk, HSC quiescence assay, ROS measurement, short- and long-term hematopoietic repopulation assay |
Stem cells |
Medium |
23939973
|
| 2016 |
Wnt5a signals through the Ryk receptor in vascular endothelial cells to activate ROCK, leading to phosphorylation of LIMK2 and inactivation of cofilin-1, resulting in enhanced actin stress fiber formation and increased endothelial permeability. Ryk silencing completely prevents Wnt5a-induced endothelial hyperpermeability. WIF1 (which specifically interferes with the WIF domain of Ryk) prevents actin polymerization. |
siRNA knockdown of Ryk, trans-endothelial resistance measurement (live), ROCK/LIMK2/cofilin phosphorylation assays, WIF1 antagonist, cytoskeletal live-cell imaging |
Cell adhesion & migration |
Medium |
27159116
|
| 2018 |
Wnt5a induces dormancy of prostate cancer cells in bone via induction of SIAH2 expression, which represses Wnt/beta-catenin signaling. This dormancy effect depends on ROR2, as ROR2 knockdown abolishes Wnt5a-induced dormancy both in vitro and in vivo. |
ROR2 siRNA knockdown, SIAH2 expression analysis, beta-catenin activity reporter, in vitro dormancy assay, in vivo bone metastasis mouse model |
The Journal of experimental medicine |
Medium |
30593464
|
| 2021 |
RNF43, an E3 ubiquitin ligase, negatively regulates Wnt5a-induced noncanonical signaling. RNF43 interacts with ROR1, ROR2, VANGL1, and VANGL2. RNF43 triggers VANGL2 ubiquitination and proteasomal degradation and promotes clathrin-dependent internalization of ROR1, and inhibits ROR2 activation. These activities suppress Wnt5a-driven melanoma invasion and resistance to BRAF/MEK inhibitors. |
BioID proximity labeling, co-immunoprecipitation (RNF43 with ROR1/2/VANGL1/2), ubiquitination assay, proteasome inhibitor rescue, clathrin inhibitor, ROR2 phosphorylation assay, invasion assay, in vivo melanoma model |
eLife |
High |
34702444
|
| 2023 |
WNT5a signaling through ROR2 activates the Hippo pathway (increased phosphorylation of MST1/2, LATS1, MOB1, and YAP1) to suppress YAP1 activity and tumor growth. Deletion of MST1/2 abrogates the WNT5a response. In ROR2-negative cells, WNT5a suppresses Hippo via increased inhibitory phosphorylation of NF2/Merlin. YAP1 activity also stimulates ROR2 and WNT5a expression, forming a negative feedback loop. |
WNT5a/Foxy5 treatment, MST1/2 deletion, ROR2 expression/knockdown, phosphorylation of Hippo components by Western blot, prostate cancer organoid and xenograft models, correlation with clinical datasets |
Cancer research |
Medium |
36622276
|
| 2019 |
In melanoma cells, Wnt5a activates the NF-κB pathway through ROR1, Dvl2, and Akt (Wnt5a pathway) and TRAF2 and RIP (NF-κB pathway). Wnt5a induces IKK phosphorylation, IκB degradation, p65 phosphorylation, and p65 nuclear translocation. An autocrine Wnt5a loop is a major regulator of NF-κB and cytokine/chemokine secretion (IL-6, IL-8, IL-11, MCP-1, TNF-sRI) in melanoma. |
cDNA overexpression, RNA interference, dominant-negative mutants, NF-κB reporter assay, IκB/IKK phosphorylation Western blot, p65 nuclear translocation imaging, cytokine ELISA, endogenous Wnt5a inhibition |
Cells |
Medium |
31510045
|
| 2017 |
Wnt5a activates Daam1, which in turn activates RhoA to promote stress fiber formation and glioblastoma cell invasion. siRNA targeting Daam1 inhibits Wnt5a-induced RhoA activation, stress fiber formation, and invasion. The Wnt5a antagonist sFRP2 blocks Wnt5a-induced Daam1 and RhoA activation and invasion. |
Daam1 pull-down activation assay, RhoA activation assay, siRNA knockdown of Daam1, RhoA inhibitor (CCG-1423), sFRP2 Wnt5a antagonist, invasion assay |
Oncology reports |
Medium |
29207169
|
| 2019 |
Wnt5a promotes invasion of esophageal squamous cell carcinoma cells via ROR1 and ROR2 receptors through DAAM1/RhoA signaling. ROR1 and ROR2 associate with each other. Downstream of ROR1/ROR2, DAAM1 activates RhoA (not Rac1/Rac2), and RhoA promotes actin filament reorganization and invasion. DAAM1 shRNA inhibits RhoA activation, which is rescued by wild-type DAAM1 overexpression. |
Co-immunoprecipitation (ROR1-ROR2), DAAM1 activity pull-down, RhoA/Rac activation assay, shRNA knockdown, wild-type DAAM1 rescue, RhoA inhibitor CCG-1423, invasion assay, phalloidin actin staining |
Cancer management and research |
Medium |
31114334
|
| 2015 |
Wnt5a inhibits hepatocyte proliferation and concludes beta-catenin signaling during liver regeneration via the Wnt5a/Frizzled-2 axis. Treatment of primary hepatocytes with Wnt5a decreases beta-catenin-TCF4 activity and cyclin-D1 expression and reduces proliferation. Liver-specific Wntless knockout (impairing Wnt secretion) reduces Wnt5a secretion and prolongs hepatocyte proliferation with increased beta-catenin/TCF4 interaction. |
Liver-specific Wntless knockout mice, partial hepatectomy model, Wnt5a treatment of primary hepatocytes, beta-catenin/TCF4 co-immunoprecipitation, cyclin-D1 Western blot, BrdU proliferation assay |
The American journal of pathology |
Medium |
26100214
|
| 2020 |
Wnt5a promotes lysosomal cholesterol egress by binding to NPC1 and NPC2 lysosomal cholesterol transporters, increasing lysosomal acid lipase expression, and decreasing mTORC1 signaling. Loss of Wnt5a in vascular smooth muscle cells (CRISPR/Cas9 or conditional knockout) disrupts lysosomal function, decreases cholesterol in the endoplasmic reticulum, and promotes atherosclerosis. |
Conditional Wnt5a knockout in vascular smooth muscle cells (mouse), CRISPR/Cas9 Wnt5a knockout in human cells, NPC1/NPC2 binding assay, lysosomal acid lipase expression, mTORC1 activity assay, cholesterol trafficking assay |
Circulation research |
High |
34886684
|
| 2020 |
High-salt diet increases vascular Wnt5a expression, which is required for Ang II-induced RhoA/ROCK activation (measured by p-MYPT1). In cultured vascular smooth muscle cells, siRNA knockdown of Wnt5a abolishes Ang II-induced p-MYPT1 upregulation. Klotho supplementation inhibits Wnt5a- and Ang II-induced p-MYPT1. Wnt5a antagonist Box5 and Wnt inhibitor LGK974 reduce salt-induced hypertension similarly to ROCK inhibitor fasudil. |
Wnt5a siRNA in VSMCs, pharmacological Wnt5a antagonist Box5, Klotho supplementation, p-MYPT1 as RhoA/ROCK activity readout, blood pressure measurement, renal blood flow assay, Klotho-knockout and aged mouse models |
The Journal of clinical investigation |
Medium |
32597829
|
| 2019 |
STAT3 transcriptionally activates Wnt5a in CLL cells by directly binding to the Wnt5a gene promoter. ChIP and EMSA confirmed STAT3 binding to four putative STAT3 binding sites near the Wnt5a promoter. STAT3 shRNA reduces Wnt5a mRNA and protein levels, and Wnt5a siRNA increases spontaneous apoptosis of CLL cells. |
ChIP of STAT3 at Wnt5a promoter, EMSA, luciferase reporter assay, STAT3 shRNA, Wnt5a siRNA, apoptosis assay, flow cytometry |
Journal of immunology |
Medium |
31645416
|
| 2023 |
PRMT2 mediates H3R8 asymmetric dimethylation (H3R8me2a) at the WNT5A promoter to enhance WNT5A transcriptional expression, leading to activation of Wnt signaling and promotion of RCC cell proliferation and motility. ChIP confirmed H3R8me2a enrichment at the WNT5A promoter by PRMT2. |
ChIP for H3R8me2a at WNT5A promoter, PRMT2 overexpression and knockdown, in vitro proliferation and motility assays, in vivo xenograft model |
Cell death & disease |
Medium |
37173306
|
| 2019 |
Wnt5a transcription in chondrocytes is activated by transcription factors Trps1 and Gli3A (activator form). ChIP and reporter gene assays identified two upstream regulatory sequences (URS) in the Wnt5a promoter; URS1 is bound by Trps1 and Gli3A in vitro and in vivo. Loss of both transcription factors decreases endogenous Wnt5a mRNA and protein levels during chondrocyte differentiation. |
ChIP of Trps1 and Gli3A at Wnt5a promoter URS1, reporter gene assay, Trps1/Gli3 mutant mouse models (double mutants), qRT-PCR and Western blot of Wnt5a |
Developmental biology |
Medium |
31550480
|
| 2020 |
In PGC1alpha-silenced melanoma, TCF12 is activated and increases WNT5A expression. WNT5A in turn stabilizes YAP protein levels to promote melanoma migration and metastasis. Inhibition of TCF12, WNT5A, or YAP individually blocks melanoma migration in vitro and metastasis in vivo. |
EZH2 inhibitor (H3K27me3 removal), TCF12 knockdown/overexpression, WNT5A inhibition, YAP inhibition, in vitro migration assay, in vivo metastasis assay |
The Journal of clinical investigation |
Medium |
31929186
|
| 2022 |
In HIV gp120-induced pain, neurons secrete Wnt5a which signals to astrocytes via ROR2 to induce astrogliosis. Conditional knockout of Wnt5a in neurons or ROR2 in astrocytes abolishes gp120-induced astrogliosis, mechanical hyperalgesia, and neural circuit polarization. Astrogliosis promotes hyperalgesia via IL-1beta regulated by a Wnt5a-ROR2-MMP2 axis. |
Conditional knockout of Wnt5a in neurons and ROR2 in astrocytes, gp120 mouse pain model, IL-1beta measurement, MMP2 inhibition, mechanical hyperalgesia testing |
Brain |
Medium |
35040478
|
| 2021 |
In the fourth ventricle choroid plexus, WNT5A is expressed by Foxj1-positive epithelial cells at villous tips and acts locally to activate non-canonical Wnt signaling via ROR1 and ROR2 receptors. Conditional Wnt5a deletion in these epithelial cells impairs branched villous morphology. MEIS1 transcription factor binds the proximal Wnt5a promoter and regulates Wnt5a expression in 4V ChP development. |
Conditional Wnt5a knockout (Foxj1-Cre), systemic Wnt5a knockout and overexpression, ChIP of MEIS1 at Wnt5a promoter, MEIS gain/loss-of-function, immunofluorescence for ROR1/ROR2 activation |
Development |
Medium |
34032267
|
| 2020 |
Conditional Wnt5a loss-of-function (Wnt5aCAG) during saccular/alveolar lung development reveals that Wnt5a is required for myofibroblast differentiation and migration but not endothelial or epithelial differentiation in alveologenesis. Conditional inactivation of both Ror1 and Ror2 in alveolar myofibroblasts recapitulates the Wnt5aCAG phenotype, demonstrating that myofibroblast defects are the primary cause of arrested alveologenesis downstream of Wnt5a. |
Conditional Wnt5a knockout (Wnt5aCAG), conditional Ror1/Ror2 double knockout in myofibroblasts, histological and immunofluorescence analysis, in vitro isolated lung fibroblast assays |
Cells |
Medium |
32046118
|
| 2022 |
Microglial WNT5A is the most abundant Wnt expressed in microglia and promotes neuronal spine maturation. Co-culture of microglia with neurons increases spine density and reduces turnover; silencing microglial Wnt5a diminishes these effects. Microglial Wnt5a knockdown reduces PSD95 levels, synaptic density, and neuronal firing rate. |
shRNA silencing of Wnt5a in microglia, co-culture with Thy1-YFP+ neurons, dendritic spine imaging (density, turnover), PSD95/VGLUT2 co-localization, multi-electrode array neuronal firing |
Brain, behavior, and immunity |
Medium |
36395958
|