The Experts below are selected from a list of 7848 Experts worldwide ranked by ideXlab platform
Akira Kikuchi - One of the best experts on this subject based on the ideXlab platform.
-
Wnt5b-associated exosomes promote cancer cell migration and proliferation
Cancer Science, 2017Co-Authors: Takeshi Harada, Chihiro Awada, Shosei Kishida, Michiko Kishida, Toshifumi Takao, Hideki Yamamoto, Akira KikuchiAbstract:© 2016 The Authors. Cancer Science published by John Wiley & Sons Australia, Ltd on behalf of Japanese Cancer Association. Wnt5b is a member of the same family of proteins as WNT5A, the overexpression of which is associated with cancer aggressiveness. Wnt5b is also suggested to be involved in cancer progression, however, details remain unclarified. We analyzed the biochemical properties of purified Wnt5b and the mode of secretion of Wnt5b by cancer cells. Wnt5b was glycosylated at three asparagine residues and lipidated at one serine residue, and these post-translational modifications of Wnt5b were essential for secretion. Purified Wnt5b showed Dvl2 phosphorylation and Rac activation abilities to a similar extent as WNT5A. In cultured-cell conditioned medium, Wnt5b was detected in supernatant or precipitation fractions that were separated by centrifugation at 100 000 g. In PANC-1 pancreatic cancer cells, 55% of secreted endogenous Wnt5b was associated with exosomes. Exosomes from wild-type PANC-1 cells, but not those from Wnt5b-knockout PANC-1 cells, activated Wnt5b signaling in CHO cells and stimulated migration and proliferation of A549 lung adenocarcinoma cells, suggesting that endogenous, Wnt5b-associated exosomes are active. The exosomes were taken up by CHO cells and immunoelectron microscopy revealed that Wnt5b is indeed associated with exosomes. In Caco-2 colon cancer cells, most Wnt5b was recovered in precipitation fractions when Wnt5b was ectopically expressed (Caco-2/Wnt5b cells). Knockdown of TSG101, an exosome marker, decreased the secretion of Wnt5b-associated exosomes from Caco-2/Wnt5b cells and inhibited Wnt5b-dependent cell proliferation. Exosomes secreted from Caco-2/Wnt5b cells stimulated migration and proliferation of A549 cells. These results suggest that Wnt5b-associated exosomes promote cancer cell migration and proliferation in a paracrine manner.
-
an anti WNT5A antibody suppresses metastasis of gastric cancer cells in vivo by inhibiting receptor mediated endocytosis
Molecular Cancer Therapeutics, 2012Co-Authors: Hideaki Hanaki, Hideki Yamamoto, Akira Sato, Hiroshi Sakane, Shinji Matsumoto, Hideki Ohdan, Akira KikuchiAbstract:WNT5A is a representative ligand that activates the β-catenin–independent pathway in Wnt signaling. It was reported that the expression of WNT5A in human gastric cancer is associated with aggressiveness and poor prognosis and that knockdown of WNT5A reduces the ability of gastric cancer cells to metastasize in nude mice. Therefore, WNT5A and its signaling pathway might be important targets for the therapy of gastric cancer. The aim of this study was to examine whether an anti-WNT5A antibody affects metastasis of gastric cancer cells. One anti-WNT5A polyclonal antibody (pAb5a-5) inhibited migration and invasion activities in vitro of gastric cancer cells with a high expression level of WNT5A. Previously, it was shown that WNT5A induces the internalization of receptors, which is required for WNT5A-dependent activation of Rac1. pAb5a-5 inhibited WNT5A-dependent internalization of receptors, thereby suppressed WNT5A-dependent activation of Rac1. Laminin γ2 is one of target genes of WNT5A signaling and Rac1 was involved in its expression. pAb5a-5 also inhibited WNT5A-dependent expression of laminin γ2. In an experimental liver metastasis assay, gastric cancer cells were introduced into the spleens of nude mice. Laminin γ2 was required for liver metastatic ability of gastric cancer cells in vivo . Furthermore, intraperitoneal injection of pAb5a-5 inhibited the metastatic ability of gastric cancer cells. These results suggest that an anti-WNT5A antibody was capable of suppressing WNT5A-dependent internalization of receptors, resulting in the prevention of metastasis of gastric cancer cells by inhibiting the activation of Rac1 and the expression of laminin γ2. Mol Cancer Ther; 11(2); 298–307. ©2011 AACR .
-
Localization of glypican-4 in different membrane microdomains is involved in the regulation of Wnt signaling.
Journal of cell science, 2012Co-Authors: Hiroshi Sakane, Hideki Yamamoto, Akira Sato, Shinji Matsumoto, Akira KikuchiAbstract:Glypicans are members of the heparan sulfate proteoglycans (HSPGs) and are involved in various growth factor signaling mechanisms. Although HSPGs affect the β-catenin-dependent and -independent pathways of Wnt signaling, how they regulate distinct Wnt pathways is not clear. It has been suggested that the β-catenin-dependent pathway is initiated through receptor endocytosis in lipid raft microdomains and the independent pathway is activated through receptor endocytosis in non-lipid raft microdomains. Here, evidence is presented that glypican-4 (GPC4) is localized to both membrane microdomains and that the localization affects its ability to regulate distinct Wnt pathways. GPC4 bound to Wnt3a and WNT5A, which activate the β-catenin-dependent and -independent pathways, respectively, and colocalized with Wnts on the cell surface. LRP6, one of Wnt3a coreceptors, was present in lipid raft microdomains, whereas Ror2, one of WNT5A coreceptors, was localized to non-lipid raft microdomains. Expression of GPC4 enhanced the Wnt3a-dependent β-catenin pathway and the WNT5A-dependent β-catenin-independent pathway, and knockdown of GPC4 suppressed both pathways. A GPC4 mutant that was localized to only non-lipid raft microdomains inhibited the β-catenin-dependent pathway but enhanced the β-catenin-independent pathway. These results suggest that GPC4 concentrates Wnt3a and WNT5A to the vicinity of their specific receptors in different membrane microdomains, thereby regulating distinct Wnt signaling.
-
WNT5A its signalling functions and implication in diseases
Acta Physiologica, 2012Co-Authors: Akira Kikuchi, Akira Sato, Hirofumi Yamamoto, Shinji MatsumotoAbstract:: WNT5A is a representative ligand that activates the β-catenin-independent pathways. Because the β-catenin-independent pathway includes multiple signalling cascades in addition to the planar cell polarity and Ca(2+) pathway, WNT5A regulates a variety of cellular functions, such as proliferation, differentiation, migration, adhesion and polarity. Consistent with the multiple functions of WNT5A signalling, WNT5A knockout mice show various phenotypes, including an inability to extend the embryonic anterior-posterior and proximal-distal axes in outgrowth tissues. Thus, many important roles of WNT5A in developmental processes have been demonstrated. Moreover, recent reports suggest that the postnatal abnormalities in the WNT5A signalling are involved in various diseases, such as cancer, inflammatory diseases and metabolic disorders. Therefore, WNT5A and its signalling pathways could be important targets for the diagnosis and therapy for human diseases.
-
ror2 frizzled complex mediates WNT5A induced ap 1 activation by regulating dishevelled polymerization
Molecular and Cellular Biology, 2010Co-Authors: Michiru Nishita, Akira Kikuchi, Akira Nomachi, Shinji Takada, Daisuke Inaba, Mitsuharu Endo, Sumiyo Itsukushima, Zhichao Wang, Sen Qiao, Yasuhiro MinamiAbstract:The receptor tyrosine kinase Ror2 acts as a receptor or coreceptor for WNT5A to mediate WNT5A-induced activation of the Wnt/JNK pathway and inhibition of the β-catenin-dependent canonical Wnt pathway. However, little is known about how Ror2 cooperates with another receptor component(s) to mediate WNT5A signaling. We show here that Ror2 regulates WNT5A-induced polymerization of Dishevelled (Dvl) and that this Ror2-mediated regulation of Dvl is independent of the cytoplasmic region of Ror2. Ror2 can associate with Frizzled7 (Fz7) via its extracellular cysteine-rich domain to form a receptor complex that is required for the regulation of Dvl and activation of the AP-1 promoter after WNT5A stimulation. Suppressed expression of Fz7 indeed results in the inhibition of WNT5A-induced polymerization of Dvl and AP-1 activation. Interestingly, both the DIX and the DEP domains of Dvl are indispensable for Dvl polymerization and subsequent AP-1 activation after WNT5A stimulation. We further show that polymerized Dvl is colocalized with Rac1 and that suppressed expression of Rac1 inhibits WNT5A-induced AP-1 activation. Collectively, our results indicate that Ror2/Fz receptor complex plays an important role in the WNT5A/Rac1/AP-1 pathway by regulating the polymerization of Dvl.
Yasuhiro Minami - One of the best experts on this subject based on the ideXlab platform.
-
the WNT5A ror2 axis promotes the signaling circuit between interleukin 12 and interferon γ in colitis
Scientific Reports, 2015Co-Authors: Akira Sato, Hirofumi Koyama, Shinji Matsumoto, Yasuhiro Minami, Kensaku Shojima, Hisako Kayama, Satoshi Nojima, Eiichi Morii, Hiroaki Honda, Kiyoshi TakedaAbstract:WNT5A, which regulates various cellular functions in Wnt signaling, is involved in inflammatory responses, however the mechanism is not well understood. We examined the role of WNT5A signaling in intestinal immunity using conditional knockout mice for WNT5A and its receptor Ror2. Removing WNT5A or Ror2 in adult mice suppressed dextran sodium sulfate (DSS)-induced colitis. It also attenuated the DSS-dependent increase in inflammatory cytokine production and decreased interferon-γ (IFN-γ)-producing CD4+ Th1 cell numbers in the colon. WNT5A was highly expressed in stromal fibroblasts in ulcerative lesions in the DSS-treated mice and inflammatory bowel disease patients. Dendritic cells (DCs) isolated from the colon of WNT5A and Ror2 deficient mice reduced the ability to differentiate naive CD4+ T cells to IFN-γ-producing CD4+ Th1 cells. In vitro experiments demonstrated that the WNT5A-Ror2 signaling axis augmented the DCs priming effect of IFN-γ, leading to enhanced lipopolysaccharide (LPS)-induced interleukin (IL)-12 expression. Taken together, these results suggest that WNT5A promotes IFN-γ signaling, leading to IL-12 expression in DCs, and thereby inducing Th1 differentiation in colitis.
-
insight into the role of WNT5A induced signaling in normal and cancer cells
International Review of Cell and Molecular Biology, 2015Co-Authors: Mitsuharu Endo, Michiru Nishita, Masanori Fujii, Yasuhiro MinamiAbstract:Abstract WNT5A is involved in the activation of noncanonical Wnt signaling, including planar cell polarity (PCP) and Wnt-Ca 2+ pathways. The Ror-family of receptor tyrosine kinases is composed of Ror1 and Ror2 in mammals. Ror2 acts as a receptor or coreceptor for WNT5A and regulates WNT5A-induced activation of PCP pathway, and WNT5A–Ror2 axis indeed plays critical roles in the developmental morphogenesis by regulating cell polarity and migration. Furthermore, WNT5A–Ror2 axis is constitutively activated in cancer cells and confers highly motile and invasive properties on cancer cells through the expression of matrix metalloproteinase genes and enhanced formation of invadopodia. Meanwhile, WNT5A also exhibits a tumor-suppressive function in certain cancers, including breast and colorectal carcinomas. Thus, it is of great importance to understand the respective molecular mechanisms governing WNT5A-mediated tumor-progressive and tumor-suppressive functions, in order to develop novel and proper diagnostic and therapeutic strategies targeting WNT5A signaling for human cancers.
-
ror2 frizzled complex mediates WNT5A induced ap 1 activation by regulating dishevelled polymerization
Molecular and Cellular Biology, 2010Co-Authors: Michiru Nishita, Akira Kikuchi, Akira Nomachi, Shinji Takada, Daisuke Inaba, Mitsuharu Endo, Sumiyo Itsukushima, Zhichao Wang, Sen Qiao, Yasuhiro MinamiAbstract:The receptor tyrosine kinase Ror2 acts as a receptor or coreceptor for WNT5A to mediate WNT5A-induced activation of the Wnt/JNK pathway and inhibition of the β-catenin-dependent canonical Wnt pathway. However, little is known about how Ror2 cooperates with another receptor component(s) to mediate WNT5A signaling. We show here that Ror2 regulates WNT5A-induced polymerization of Dishevelled (Dvl) and that this Ror2-mediated regulation of Dvl is independent of the cytoplasmic region of Ror2. Ror2 can associate with Frizzled7 (Fz7) via its extracellular cysteine-rich domain to form a receptor complex that is required for the regulation of Dvl and activation of the AP-1 promoter after WNT5A stimulation. Suppressed expression of Fz7 indeed results in the inhibition of WNT5A-induced polymerization of Dvl and AP-1 activation. Interestingly, both the DIX and the DEP domains of Dvl are indispensable for Dvl polymerization and subsequent AP-1 activation after WNT5A stimulation. We further show that polymerized Dvl is colocalized with Rac1 and that suppressed expression of Rac1 inhibits WNT5A-induced AP-1 activation. Collectively, our results indicate that Ror2/Fz receptor complex plays an important role in the WNT5A/Rac1/AP-1 pathway by regulating the polymerization of Dvl.
-
cell tissue tropic functions of WNT5A signaling in normal and cancer cells
Trends in Cell Biology, 2010Co-Authors: Michiru Nishita, Masahiro Enomoto, Kaoru Yamagata, Yasuhiro MinamiAbstract:Correct spatio-temporal regulation of WNT5A signaling is required for normal developmental morphogenesis, and defects in this pathway are linked to tumorigenesis. The precise role of WNT5A signaling in cancer has, however, been a matter of controversy. Loss of WNT5A signaling is related to development of lymphoid malignancies, whereas constitutively active WNT5A signaling is involved in invasion or metastasis of several cancers. Interestingly, recent studies in Drosophila and mouse have revealed that disrupted cell polarity might contribute to invasion/metastasis of cancers. WNT5A activates the planar cell polarity (PCP) pathway, partly through the receptor tyrosine kinase Ror2. Here, we review developments in our understanding of the molecular mechanisms underlying WNT5A signaling, with an emphasis on the role of Ror2 in cancer. We also propose a model where the outcomes of normal and aberrant WNT5A/Ror2 signaling depend on cell/tissue-tropic contexts.
-
receptor tyrosine kinase ror2 mediates WNT5A induced polarized cell migration by activating c jun n terminal kinase via actin binding protein filamin a
Journal of Biological Chemistry, 2008Co-Authors: Akira Nomachi, Michiru Nishita, Daisuke Inaba, Masahiro Enomoto, Mayumi Hamasaki, Yasuhiro MinamiAbstract:Abstract The receptor tyrosine kinase Ror2 has recently been shown to act as an alternative receptor or coreceptor for WNT5A and to mediate WNT5A-induced migration of cultured cells. However, little is known about the molecular mechanism underlying this migratory process. Here we show by wound-healing assays that Ror2 plays critical roles in WNT5A-induced cell migration by regulating formation of lamellipodia and reorientation of microtubule-organizing center (MTOC). WNT5A stimulation induces activation of the c-Jun N-terminal kinase JNK at the wound edge in a Ror2-dependent manner, and inhibiting JNK activity abrogates WNT5A-induced lamellipodia formation and MTOC reorientation. Additionally, the association of Ror2 with the actin-binding protein filamin A is required for WNT5A-induced JNK activation and polarized cell migration. We further show that WNT5A-induced JNK activation and MTOC reorientation can be suppressed by inhibiting PKCζ. Taken together, our findings indicate that WNT5A/Ror2 activates JNK, through a process involving filamin A and PKCζ, to regulate polarized cell migration.
Masaru Katoh - One of the best experts on this subject based on the ideXlab platform.
-
transcriptional mechanisms of WNT5A based on nf κb hedgehog tgfβ and notch signaling cascades
International Journal of Molecular Medicine, 2009Co-Authors: Masuko Katoh, Masaru KatohAbstract:: WNT5A is a cancer-associated gene involved in invasion and metastasis of melanoma, breast cancer, pancreatic cancer, and gastric cancer. WNT5A transduces signals through Frizzled, ROR1, ROR2 or RYK receptors to beta-catenin-TCF/LEF, DVL-RhoA-ROCK, DVL-RhoB-Rab4, DVL-Rac-JNK, DVL-aPKC, Calcineurin-NFAT, MAP3K7-NLK, MAP3K7-NF-kappaB, and DAG-PKC signaling cascades in a context-dependent manner. SNAI1 (Snail), CD44, G3BP2, and YAP1 are WNT5A target genes. We and other groups previously reported that IL6- or LIF-induced signaling through JAK-STAT3 signaling cascade is involved in WNT5A upregulation (STAT3-WNT5A signaling loop). Here, refined integrative genomic analyses of WNT5A were carried out to elucidate other mechanisms of WNT5A transcription. The WNT5A gene was found to encode two isoforms by using alternative first exons 1A and 1B. Quadruple Smad-binding elements (SBEs), single Sp1-binding site (GC-box), PPARgamma-binding site, C/EBP-binding site and bHLH-binding site within the promoter A region, 5'-adjacent to exon 1A, were conserved in human WNT5A, chimpanzee WNT5A, mouse WNT5A, and rat WNT5A. NF-kappaB-binding site, CUX1-binding site, double SBEs and double GC-boxes within the promoter B region, 5'-adjacent to exon 1B, were conserved in mammalian WNT5A orthologs. Quadruple FOX-binding sites and double SBEs within ultra-conserved intron 1 were also conserved in mammalian WNT5A orthologs. Conserved NF-kappaB-binding site within the WNT5A promoter B region elucidated the mechanisms that TNFalpha and toll-like receptor (TLR) signals upregulate WNT5A via MAP3K7. Quadruple FOX-binding sites rather than GLI-binding site revealed that Hedgehog signals induce WNT5A upregulation indirectly via FOX family members, such as FOXA2, FOXC2, FOXE1, FOXF1 and FOXL1. TGFbeta signals were found to upregulate WNT5A expression directly through the Smad complex, and also indirectly through Smad-induced CUX1 and MAP3K7-mediated NF-kappaB. Together these facts indicate that WNT5A is transcribed based on multiple mechanisms, such as NF-kappaB, Hedgehog, TGFbeta, and Notch signaling cascades.
-
stat3 induced WNT5A signaling loop in embryonic stem cells adult normal tissues chronic persistent inflammation rheumatoid arthritis and cancer review
International Journal of Molecular Medicine, 2007Co-Authors: Masuko Katoh, Masaru KatohAbstract:: Leukemia inhibitory factor (LIF), oncostatin M, leptin, ciliary neurotrophic factor, cardiotrophin 1, cardiotrophin-like cytokine factor 1, interleukin 6 (IL6), interleukin 11 and interleukin 27 activate the gp130-JAK-STAT3 signaling cascade. Here, WNT5A was characterized as the evolutionarily conserved target of the STAT3 signaling cascade based on 11-bp-spaced tandem STAT3-binding sites within intron 4 of human, chimpanzee, cow, mouse and rat WNT5A orthologs. Canonical WNT5A signaling through Frizzled and LRP5/LRP6 receptors activates FGF20, WISP1, MYC and CCND1 transcription for the maintenance of stem/progenitor cells, while non-canonical WNT5A signaling through Frizzled and ROR2/PTK7/RYK receptors activates the RHOA, JNK, NLK and NFAT signaling cascades for the control of tissue polarity, cell adhesion or movement. LIF-induced WNT5A activates canonical Wnt signaling in mouse embryonic stem cells for self-renewal. STAT3-induced WNT5A activates non-canonical Wnt signaling in rat cardiac myocytes for N-cadherin-dependent aggregation. IL6, secreted from epithelial cells or macrophages, induces WNT5A upregulation in mesenchymal cells. WNT5A then activates canonical WNT signaling in epithelial cells. IL6-induced WNT5A activates canonical WNT signaling for autocrine proliferation of human synovial fibroblasts in rheumatoid arthritis. IL-6 signaling is activated during human chronic atrophic gastritis with Helicobacter pylori infection, and aberrant Stat3 signaling activation gives rise to mouse gastric tumors. WNT5A is frequently upregulated in human primary gastric cancer due to tumor-stromal interaction. WNT5A might be downregulated in advanced cancer with poorer prognosis due to genetic alterations compensating WNT5A signaling. Oncogenic WNT5A activates canonical WNT signaling in cancer stem cells for self-renewal, and non-canonical WNT signaling at the tumor-stromal interface for invasion and metastasis. SNP of genes encoding components of the cytokine-induced WNT5A signaling loop is a predicted risk factor for RA and cancer, especially diffuse-type gastric and pancreatic cancer. Humanized anti-IL6 receptor antibody and WNT5A mimetic small-molecule antagonist could be applied to personalized medicine for RA and cancer driven by the IL6-induced WNT5A signaling loop.
-
Networking of WNT, FGF, Notch, BMP, and Hedgehog Signaling Pathways during Carcinogenesis
Stem Cell Reviews, 2007Co-Authors: Masaru KatohAbstract:The biological functions of some orthologs within the human genome and model-animal genomes are evolutionarily conserved, but those of others are divergent due to protein evolution and promoter evolution. Because WNT signaling molecules play key roles during embryogenesis, tissue regeneration and carcinogenesis, the author’s group has carried out a human WNT-ome project for the comprehensive characterization of human genes encoding WNT signaling molecules. From 1996 to 2002, we cloned and characterized WNT2B/WNT13, WNT3, WNT3A, WNT5B, WNT6, WNT7B, WNT8A, WNT8B, WNT9A/WNT14, WNT9B/WNT14B, WNT10A, WNT10B, WNT11, FZD1, FZD2, FZD3, FZD4, FZD5, FZD6, FZD7, FZD8, FZD10, FRAT1, FRAT2, NKD1, NKD2, VANGL1, RHOU/ARHU, RHOV/ARHV, GIPC2, GIPC3, FBXW11/βTRCP2, SOX17, TCF7L1/TCF3 , and established a cDNA-PCR system for snap-shot and dynamic analyses on the WNT-transcriptome. In 2003, we identified and characterized PRICKLE1, PRICKLE2, DACT1/DAPPER1, DACT2/DAPPER2, DAAM2 , and BCL9L . After completion of the human WNT-ome project, we have been working on the stem cell signaling network. WNT signals are transduced to β-catenin, NLK, NFAT, PKC, JNK and RhoA signaling cascades. FGF20, JAG1 and DKK1 are target genes of the WNT-β-catenin signaling cascade. Cross-talk of WNT and FGF signaling pathways potentiates β-catenin and NFAT signaling cascades. BMP signals induce IHH upregulation in co-operation with RUNX. Hedgehog signals induce upregulation of SFRP1, JAG2 and FOXL1 , and then FOXL1 induces BMP4 upregulation. The balance between WNT-FGF-Notch and BMP-Hedgehog signaling networks is important for the maintenance of homoestasis among stem and progenitor cells. Disruption of the stem cell signaling network results in pathological conditions, such as congenital diseases and cancer.
-
Comparative genomics on WNT5A and Wnt5b genes
International Journal of Molecular Medicine, 2005Co-Authors: Masuko Katoh, Masaru KatohAbstract:: Canonical WNTs (WNT2, WNT2B, etc) activate the beta-catenin-TCF pathway to induce carcinogenesis, while non-canonical WNTs (WNT5A, WNT11, etc) activate the planar cell polarity (PCP) pathway to induce cell motility and metastasis. WNT5A gene at chromosome 3p14.3 and WNT5B gene at chromosome 12p13.33 are paralogs within the human genome. Here, we identified and characterized rat WNT5A and Wnt5b genes by using bioinformatics. Rat WNT5A and Wnt5b genes, consisting of five exons, were identified within AC095764.5 and AC112027.3 genome sequences, respectively. Rat WNT5A (380 aa) and Wnt5b (359 aa) were secreted proteins with 24 conserved Cys residues and four Asn-linked glycosylation sites, which showed 75.8% total-amino-acid identity. Nucleotide position 182586-183836 of AC095764.5 genome sequence and nucleotide position 161044-159886 of AC121764.2 genome sequence were identified as evolutionarily conserved rat WNT5A and human WNT5A promoters, respectively. Nucleotide identity between rat WNT5A and human WNT5A promoters was 72.5%. E47 and NKX2-5-binding sites were evolutionarily conserved among rat WNT5A, mouse WNT5A, and human WNT5A promoters. On the other hand, rodent Wnt5b promoters and human WNT5B promoter were significantly divergent. Up-regulation of Wnt5b during rodent adipocytic differentiation does not simply indicate the implication of WNT5B in human adipogenesis. Real susceptibility gene for type 2 diabetes, associated with SNP within intron 3 of human WNT5B gene (IMS-JST024404), remains to be identified. This is the first report on rat WNT5A and Wnt5b genes as well as on comparative genomics for WNT5A and Wnt5b orthologs.
Michiru Nishita - One of the best experts on this subject based on the ideXlab platform.
-
insight into the role of WNT5A induced signaling in normal and cancer cells
International Review of Cell and Molecular Biology, 2015Co-Authors: Mitsuharu Endo, Michiru Nishita, Masanori Fujii, Yasuhiro MinamiAbstract:Abstract WNT5A is involved in the activation of noncanonical Wnt signaling, including planar cell polarity (PCP) and Wnt-Ca 2+ pathways. The Ror-family of receptor tyrosine kinases is composed of Ror1 and Ror2 in mammals. Ror2 acts as a receptor or coreceptor for WNT5A and regulates WNT5A-induced activation of PCP pathway, and WNT5A–Ror2 axis indeed plays critical roles in the developmental morphogenesis by regulating cell polarity and migration. Furthermore, WNT5A–Ror2 axis is constitutively activated in cancer cells and confers highly motile and invasive properties on cancer cells through the expression of matrix metalloproteinase genes and enhanced formation of invadopodia. Meanwhile, WNT5A also exhibits a tumor-suppressive function in certain cancers, including breast and colorectal carcinomas. Thus, it is of great importance to understand the respective molecular mechanisms governing WNT5A-mediated tumor-progressive and tumor-suppressive functions, in order to develop novel and proper diagnostic and therapeutic strategies targeting WNT5A signaling for human cancers.
-
ror2 frizzled complex mediates WNT5A induced ap 1 activation by regulating dishevelled polymerization
Molecular and Cellular Biology, 2010Co-Authors: Michiru Nishita, Akira Kikuchi, Akira Nomachi, Shinji Takada, Daisuke Inaba, Mitsuharu Endo, Sumiyo Itsukushima, Zhichao Wang, Sen Qiao, Yasuhiro MinamiAbstract:The receptor tyrosine kinase Ror2 acts as a receptor or coreceptor for WNT5A to mediate WNT5A-induced activation of the Wnt/JNK pathway and inhibition of the β-catenin-dependent canonical Wnt pathway. However, little is known about how Ror2 cooperates with another receptor component(s) to mediate WNT5A signaling. We show here that Ror2 regulates WNT5A-induced polymerization of Dishevelled (Dvl) and that this Ror2-mediated regulation of Dvl is independent of the cytoplasmic region of Ror2. Ror2 can associate with Frizzled7 (Fz7) via its extracellular cysteine-rich domain to form a receptor complex that is required for the regulation of Dvl and activation of the AP-1 promoter after WNT5A stimulation. Suppressed expression of Fz7 indeed results in the inhibition of WNT5A-induced polymerization of Dvl and AP-1 activation. Interestingly, both the DIX and the DEP domains of Dvl are indispensable for Dvl polymerization and subsequent AP-1 activation after WNT5A stimulation. We further show that polymerized Dvl is colocalized with Rac1 and that suppressed expression of Rac1 inhibits WNT5A-induced AP-1 activation. Collectively, our results indicate that Ror2/Fz receptor complex plays an important role in the WNT5A/Rac1/AP-1 pathway by regulating the polymerization of Dvl.
-
cell tissue tropic functions of WNT5A signaling in normal and cancer cells
Trends in Cell Biology, 2010Co-Authors: Michiru Nishita, Masahiro Enomoto, Kaoru Yamagata, Yasuhiro MinamiAbstract:Correct spatio-temporal regulation of WNT5A signaling is required for normal developmental morphogenesis, and defects in this pathway are linked to tumorigenesis. The precise role of WNT5A signaling in cancer has, however, been a matter of controversy. Loss of WNT5A signaling is related to development of lymphoid malignancies, whereas constitutively active WNT5A signaling is involved in invasion or metastasis of several cancers. Interestingly, recent studies in Drosophila and mouse have revealed that disrupted cell polarity might contribute to invasion/metastasis of cancers. WNT5A activates the planar cell polarity (PCP) pathway, partly through the receptor tyrosine kinase Ror2. Here, we review developments in our understanding of the molecular mechanisms underlying WNT5A signaling, with an emphasis on the role of Ror2 in cancer. We also propose a model where the outcomes of normal and aberrant WNT5A/Ror2 signaling depend on cell/tissue-tropic contexts.
-
receptor tyrosine kinase ror2 mediates WNT5A induced polarized cell migration by activating c jun n terminal kinase via actin binding protein filamin a
Journal of Biological Chemistry, 2008Co-Authors: Akira Nomachi, Michiru Nishita, Daisuke Inaba, Masahiro Enomoto, Mayumi Hamasaki, Yasuhiro MinamiAbstract:Abstract The receptor tyrosine kinase Ror2 has recently been shown to act as an alternative receptor or coreceptor for WNT5A and to mediate WNT5A-induced migration of cultured cells. However, little is known about the molecular mechanism underlying this migratory process. Here we show by wound-healing assays that Ror2 plays critical roles in WNT5A-induced cell migration by regulating formation of lamellipodia and reorientation of microtubule-organizing center (MTOC). WNT5A stimulation induces activation of the c-Jun N-terminal kinase JNK at the wound edge in a Ror2-dependent manner, and inhibiting JNK activity abrogates WNT5A-induced lamellipodia formation and MTOC reorientation. Additionally, the association of Ror2 with the actin-binding protein filamin A is required for WNT5A-induced JNK activation and polarized cell migration. We further show that WNT5A-induced JNK activation and MTOC reorientation can be suppressed by inhibiting PKCζ. Taken together, our findings indicate that WNT5A/Ror2 activates JNK, through a process involving filamin A and PKCζ, to regulate polarized cell migration.
-
filopodia formation mediated by receptor tyrosine kinase ror2 is required for WNT5A induced cell migration
Journal of Cell Biology, 2006Co-Authors: Michiru Nishita, Akira Kikuchi, Akira Nomachi, Shuichi Kani, Nagako Sougawa, Yasutaka Ohta, Shinji Takada, Yasuhiro MinamiAbstract:The receptor tyrosine kinase Ror2 plays important roles in developmental morphogenesis. It has recently been shown that Ror2 mediates WNT5A-induced noncanonical Wnt signaling by activating the Wnt–JNK pathway and inhibiting the β-catenin–TCF pathway. However, the function of Ror2 in noncanonical Wnt signaling leading to cell migration is largely unknown. We show, using genetically different or manipulated cultured cells, that Ror2 is critical for WNT5A-induced, but not Wnt3a-induced, cell migration. Ror2-mediated cell migration requires the extracellular cysteine-rich domain (CRD), which is the binding site for WNT5A, and the cytoplasmic proline-rich domain (PRD) of Ror2. Furthermore, Ror2 can mediate filopodia formation via actin reorganization, irrespective of WNT5A, and this Ror2-mediated filopodia formation requires the actin-binding protein filamin A, which associates with the PRD of Ror2. Intriguingly, disruption of filopodia formation by suppressing the expression of either Ror2 or filamin A inhibits WNT5A-induced cell migration, indicating that Ror2-mediated filopodia formation is essential for WNT5A-induced cell migration.
Akira Sato - One of the best experts on this subject based on the ideXlab platform.
-
the WNT5A ror2 axis promotes the signaling circuit between interleukin 12 and interferon γ in colitis
Scientific Reports, 2015Co-Authors: Akira Sato, Hirofumi Koyama, Shinji Matsumoto, Yasuhiro Minami, Kensaku Shojima, Hisako Kayama, Satoshi Nojima, Eiichi Morii, Hiroaki Honda, Kiyoshi TakedaAbstract:WNT5A, which regulates various cellular functions in Wnt signaling, is involved in inflammatory responses, however the mechanism is not well understood. We examined the role of WNT5A signaling in intestinal immunity using conditional knockout mice for WNT5A and its receptor Ror2. Removing WNT5A or Ror2 in adult mice suppressed dextran sodium sulfate (DSS)-induced colitis. It also attenuated the DSS-dependent increase in inflammatory cytokine production and decreased interferon-γ (IFN-γ)-producing CD4+ Th1 cell numbers in the colon. WNT5A was highly expressed in stromal fibroblasts in ulcerative lesions in the DSS-treated mice and inflammatory bowel disease patients. Dendritic cells (DCs) isolated from the colon of WNT5A and Ror2 deficient mice reduced the ability to differentiate naive CD4+ T cells to IFN-γ-producing CD4+ Th1 cells. In vitro experiments demonstrated that the WNT5A-Ror2 signaling axis augmented the DCs priming effect of IFN-γ, leading to enhanced lipopolysaccharide (LPS)-induced interleukin (IL)-12 expression. Taken together, these results suggest that WNT5A promotes IFN-γ signaling, leading to IL-12 expression in DCs, and thereby inducing Th1 differentiation in colitis.
-
WNT5A promotes cancer cell invasion and proliferation by receptor mediated endocytosis dependent and independent mechanisms respectively
Scientific Reports, 2015Co-Authors: Kensaku Shojima, Akira Sato, Hideaki Hanaki, Ikuko Tsujimoto, Masahiro Nakamura, Kazunari Hattori, Yuji Sato, Keiji Dohi, Michinari Hirata, Hideki YamamotoAbstract:WNT5A activates the Wnt/β-catenin-independent pathway and its overexpression is associated with tumor aggressiveness enhancing invasive activity. For this action, WNT5A-induced receptor endocytosis with clathrin is required. WNT5A expression was previously believed to be associated with cancer cell motility but not proliferation. Recently, it was reported that WNT5A is also implicated in cancer cell proliferation, but the mechanism was not clear. In this study, we generated a neutralizing anti-WNT5A monoclonal antibody (mAb5A16) to investigate the mechanism by which WNT5A regulates cancer cell proliferation. WNT5A stimulated both invasion and proliferation of certain types of cancer cells, including HeLaS3 cervical cancer cells and A549 lung cancer cells although WNT5A promoted invasion but not proliferation in other cancer cells such as KKLS gastric cancer cells. mAb5A16 did not affect the binding of WNT5A to its receptor, but it suppressed WNT5A-induced receptor-mediated endocytosis. mAb5A16 inhibited invasion but not proliferation of HeLaS3 and A549 cells. WNT5A activated Src family kinases (SFKs) and WNT5A-dependent cancer cell proliferation was dependent on SFKs, yet blockade of receptor-mediated endocytosis did not affect cancer cell proliferation and SFK activity. These results suggest that WNT5A promotes invasion and proliferation of certain types of cancer cells through receptor-mediated endocytosis-dependent and -independent mechanisms, respectively.
-
an anti WNT5A antibody suppresses metastasis of gastric cancer cells in vivo by inhibiting receptor mediated endocytosis
Molecular Cancer Therapeutics, 2012Co-Authors: Hideaki Hanaki, Hideki Yamamoto, Akira Sato, Hiroshi Sakane, Shinji Matsumoto, Hideki Ohdan, Akira KikuchiAbstract:WNT5A is a representative ligand that activates the β-catenin–independent pathway in Wnt signaling. It was reported that the expression of WNT5A in human gastric cancer is associated with aggressiveness and poor prognosis and that knockdown of WNT5A reduces the ability of gastric cancer cells to metastasize in nude mice. Therefore, WNT5A and its signaling pathway might be important targets for the therapy of gastric cancer. The aim of this study was to examine whether an anti-WNT5A antibody affects metastasis of gastric cancer cells. One anti-WNT5A polyclonal antibody (pAb5a-5) inhibited migration and invasion activities in vitro of gastric cancer cells with a high expression level of WNT5A. Previously, it was shown that WNT5A induces the internalization of receptors, which is required for WNT5A-dependent activation of Rac1. pAb5a-5 inhibited WNT5A-dependent internalization of receptors, thereby suppressed WNT5A-dependent activation of Rac1. Laminin γ2 is one of target genes of WNT5A signaling and Rac1 was involved in its expression. pAb5a-5 also inhibited WNT5A-dependent expression of laminin γ2. In an experimental liver metastasis assay, gastric cancer cells were introduced into the spleens of nude mice. Laminin γ2 was required for liver metastatic ability of gastric cancer cells in vivo . Furthermore, intraperitoneal injection of pAb5a-5 inhibited the metastatic ability of gastric cancer cells. These results suggest that an anti-WNT5A antibody was capable of suppressing WNT5A-dependent internalization of receptors, resulting in the prevention of metastasis of gastric cancer cells by inhibiting the activation of Rac1 and the expression of laminin γ2. Mol Cancer Ther; 11(2); 298–307. ©2011 AACR .
-
Localization of glypican-4 in different membrane microdomains is involved in the regulation of Wnt signaling.
Journal of cell science, 2012Co-Authors: Hiroshi Sakane, Hideki Yamamoto, Akira Sato, Shinji Matsumoto, Akira KikuchiAbstract:Glypicans are members of the heparan sulfate proteoglycans (HSPGs) and are involved in various growth factor signaling mechanisms. Although HSPGs affect the β-catenin-dependent and -independent pathways of Wnt signaling, how they regulate distinct Wnt pathways is not clear. It has been suggested that the β-catenin-dependent pathway is initiated through receptor endocytosis in lipid raft microdomains and the independent pathway is activated through receptor endocytosis in non-lipid raft microdomains. Here, evidence is presented that glypican-4 (GPC4) is localized to both membrane microdomains and that the localization affects its ability to regulate distinct Wnt pathways. GPC4 bound to Wnt3a and WNT5A, which activate the β-catenin-dependent and -independent pathways, respectively, and colocalized with Wnts on the cell surface. LRP6, one of Wnt3a coreceptors, was present in lipid raft microdomains, whereas Ror2, one of WNT5A coreceptors, was localized to non-lipid raft microdomains. Expression of GPC4 enhanced the Wnt3a-dependent β-catenin pathway and the WNT5A-dependent β-catenin-independent pathway, and knockdown of GPC4 suppressed both pathways. A GPC4 mutant that was localized to only non-lipid raft microdomains inhibited the β-catenin-dependent pathway but enhanced the β-catenin-independent pathway. These results suggest that GPC4 concentrates Wnt3a and WNT5A to the vicinity of their specific receptors in different membrane microdomains, thereby regulating distinct Wnt signaling.
-
WNT5A its signalling functions and implication in diseases
Acta Physiologica, 2012Co-Authors: Akira Kikuchi, Akira Sato, Hirofumi Yamamoto, Shinji MatsumotoAbstract:: WNT5A is a representative ligand that activates the β-catenin-independent pathways. Because the β-catenin-independent pathway includes multiple signalling cascades in addition to the planar cell polarity and Ca(2+) pathway, WNT5A regulates a variety of cellular functions, such as proliferation, differentiation, migration, adhesion and polarity. Consistent with the multiple functions of WNT5A signalling, WNT5A knockout mice show various phenotypes, including an inability to extend the embryonic anterior-posterior and proximal-distal axes in outgrowth tissues. Thus, many important roles of WNT5A in developmental processes have been demonstrated. Moreover, recent reports suggest that the postnatal abnormalities in the WNT5A signalling are involved in various diseases, such as cancer, inflammatory diseases and metabolic disorders. Therefore, WNT5A and its signalling pathways could be important targets for the diagnosis and therapy for human diseases.
