Wnt signaling comprises a group of pathways emanating from the extracellular environment through cell-surface receptors into the intracellular milieu. Wnt signaling cascades can be divided into two main branches, the canonical/β-catenin pathway and the non-canonical pathways containing the Wnt/planar cell polarity and Wnt/calcium signaling. Syndecans are type I transmembrane proteoglycans with a long evolutionary history, being expressed in all Bilateria and in almost all cell types. Both Wnt pathways have been extensively studied over the past 30 years and shown to have roles during development and in a multitude of diseases. Although the first evidence for interactions between syndecans and Wnts dates back to 1997, the number of studies connecting these pathways is low, and many open questions remained unanswered. In this review, syndecan's involvement in Wnt signaling pathways as well as some of the pathologies resulting from dysregulation of the components of these pathways are summarized.

• MeSH
• lidé MeSH
• nemoc MeSH
• signální dráha Wnt * MeSH
• syndekany chemie   metabolismus   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• přehledy MeSH

Kolorektální karcinom je jednou z nejčastějších malignit v hospodářsky vyspělé části světa včetně České republiky. Na molekulárně biologické úrovni je ve většině případů jeho vznik spojený s patologicky aktivovanou kanonickou Wnt signalizací. Tato rešeršní práce si klade za cíl představit čtenářům tuto mezibuněčnou signalizační dráhu, její roli v kolorektální karcinogenezi i možná terapeutická východiska vyplývající z našich aktuálních znalostí.

Colorectal cancer is among the most frequent malignancies in the economically developed part of the world including the Czech Republic. From the molecular biological point of view, the development of colorectal cancer is in most cases linked to the pathologically activated canonical Wnt signalling pathway. The aim of this review is to present this intercellular signalling pathway, its role in the colorectal carcinogenesis and possible therapeutic implications according to our current knowledge.

• Klíčová slova
• nádorová transformace buňky,
• MeSH
• antiflogistika nesteroidní farmakologie   terapeutické užití   MeSH
• beta-katenin genetika   MeSH
• DNA vazebné proteiny genetika   MeSH
• financování organizované MeSH
• geny APC MeSH
• kolorektální nádory etiologie   farmakoterapie   genetika   MeSH
• lidé MeSH
• monoklonální protilátky farmakologie   terapeutické užití   MeSH
• nádorová transformace buněk MeSH
• proteiny Wnt fyziologie   genetika   MeSH
• signální dráha Wnt fyziologie   genetika   MeSH
• signální komplex axin genetika   MeSH
• střevní sliznice fyziologie   patofyziologie   MeSH
• transkripční faktory genetika   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• přehledy MeSH

Chemotherapy-induced neuropathic pain (CNP) is the major dose-limiting factor in cancer chemotherapy. However, the mechanisms underlying CNP remain elusive. In the present study, CNP was induced by repeated intraperitoneal injection of vincristine (VCR) into male C57BL/6J mice. VCR administration caused significant activation of Wnt/beta-catenin signaling, which led to the activation of astrocytes, microglia, the release of inflammatory cytokines tumour necrosis factor (TNF)-alpha, monocyte chemoattractant protein-1 (MCP-1) and the activation of subsequent mitogen-activated protein kinase (MAPK)/extracellular signal-regulated protein kinase (ERK) signaling pathway in CNP mice. Blocking Wnt/beta-catenin signaling by intrathecal administration of the inhibitors of Wnt response (IWR) effectively attenuated VCR-induced neuropathic pain. Furthermore, IWR inhibited the activation of astrocytes, microglia, TNF-alpha, MCP-1 and MAPK/ERK signaling in the spinal cord, which was triggered by VCR-induced Wnt/beta-catenin signaling upregulation. These results suggest that Wnt/beta-catenin signaling plays a critical role in VCR-induced neuropathic pain and provides evidence for potential interfering with Wnt/beta-catenin signaling to ameliorate VCR-induced neuropathic pain.

• MeSH
• beta-katenin metabolismus   MeSH
• fytogenní protinádorové látky škodlivé účinky   farmakologie   MeSH
• modely nemocí na zvířatech MeSH
• myši inbrední C57BL MeSH
• myši MeSH
• nádory farmakoterapie   metabolismus   patologie   MeSH
• neuralgie chemicky indukované   metabolismus   patologie   MeSH
• proteiny Wnt metabolismus   MeSH
• signální dráha Wnt * MeSH
• signální transdukce MeSH
• vinkristin škodlivé účinky   farmakologie   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

This themed section of the British Journal of Pharmacology stems from the EMBO Conference: Wnt Meeting 2016 held from the 14th to 16th September 2016 in Brno, Czech Republic. LINKED ARTICLES: This article is part of a themed section on WNT Signalling: Mechanisms and Therapeutic Opportunities. To view the other articles in this section visit http://onlinelibrary.wiley.com/doi/10.1111/bph.v174.24/issuetoc.

• MeSH
• buněčná membrána účinky léků   metabolismus   MeSH
• lidé MeSH
• nádory farmakoterapie   metabolismus   patologie   MeSH
• protinádorové látky farmakologie   MeSH
• signální dráha Wnt účinky léků   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• úvodní články MeSH
• úvodníky MeSH

The seven-transmembrane-spanning receptors of the FZD1-10 class are bound and activated by the WNT family of lipoglycoproteins, thereby inducing a complex network of signaling pathways. However, the specificity of the interaction between mammalian WNT and FZD proteins and the subsequent signaling cascade downstream of the different WNT-FZD pairs have not been systematically addressed to date. In this study, we determined the binding affinities of various WNTs for different members of the FZD family by using bio-layer interferometry and characterized their functional selectivity in a cell system. Using purified WNTs, we show that different FZD cysteine-rich domains prefer to bind to distinct WNTs with fast on-rates and slow off-rates. In a 32D cell-based system engineered to overexpress FZD2, FZD4, or FZD5, we found that WNT-3A (but not WNT-4, -5A, or -9B) activated the WNT-β-catenin pathway through FZD2/4/5 as measured by phosphorylation of LRP6 and β-catenin stabilization. Surprisingly, different WNT-FZD pairs showed differential effects on phosphorylation of DVL2 and DVL3, revealing a previously unappreciated DVL isoform selectivity by different WNT-FZD pairs in 32D cells. In summary, we present extensive mapping of WNT-FZD cysteine-rich domain interactions complemented by analysis of WNT-FZD pair functionality in a unique cell system expressing individual FZD isoforms. Differential WNT-FZD binding and selective functional readouts suggest that endogenous WNT ligands evolved with an intrinsic natural bias toward different downstream signaling pathways, a phenomenon that could be of great importance in the design of FZD-targeting drugs.

• MeSH
• beta-katenin metabolismus   MeSH
• buněčné linie MeSH
• fosforylace MeSH
• frizzled receptory metabolismus   MeSH
• mapování interakce mezi proteiny MeSH
• mapy interakcí proteinů * MeSH
• myši MeSH
• protein - isoformy metabolismus   MeSH
• proteiny Wnt metabolismus   MeSH
• signální dráha Wnt * MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, N.I.H., Intramural MeSH

The lipoglycoproteins of the mammalian WNT family induce β-catenin-dependent signaling through interaction with members of the Class Frizzled receptors and LDL receptor-related protein 5/6 (LRP5/6) albeit with unknown selectivity. The 10 mammalian Frizzleds (FZDs) are seven transmembrane (7TM) spanning receptors and have recently been classified as G protein-coupled receptors (GPCRs). This review summarizes the current knowledge about WNT/FZD selectivity and functional selectivity, the role of co-receptors for signal specification, the formation of receptor complexes as well as the kinetics and mechanisms of signal initiation with focus on WNT/β-catenin signaling. In order to exploit the true therapeutic potential of WNT/FZD signaling to treat human disease, it is clear that substantial progress in the understanding of receptor complex formation and signal specification has to precede a mechanism-based drug design targeting WNT receptors.

• MeSH
• frizzled receptory chemie   metabolismus   MeSH
• kinetika MeSH
• lidé MeSH
• ligandy MeSH
• multimerizace proteinu MeSH
• signální dráha Wnt * MeSH
• substrátová specifita MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• přehledy MeSH

High grade serous carcinoma of the ovary, fallopian tube, and peritoneum (HGSC) is the deadliest gynecological disease which results in a five-year survival rate of 30% or less. HGSC is characterized by the early and rapid development of metastases accompanied by a high frequency of ascites i.e. the pathological accumulation of fluid in peritoneum. Ascites constitute a complex tumor microenvironment and contribute to disease progression by largely unknown mechanisms. Methods: Malignant ascites obtained from HGSC patients who had undergone cytoreductive surgery were tested for their ability to induce WNT signaling in the Kuramochi cell line, a novel and clinically relevant in vitro model of HGSC. Next, cancer spheroids (the main form of metastatic cancer cells in ascites) were evaluated with respect to WNT signaling. Kuramochi cells were used to determine the role of individual WNT signaling branches in the adoption of metastatic stem cell-like behavior by HGSC cells. Furthermore, we analyzed genomic and transcriptomic data on WNT/Planar Cell Polarity (PCP) components retrieved from public cancer databases and corroborated with primary patient samples and validated antibodies on the protein level. Results: We have shown that ascites are capable of inducing WNT signaling in primary HGSC cells and HGSC cell line, Kuramochi. Importantly, patients whose ascites cannot activate WNT pathway present with less aggressive disease and a considerably better outcome including overall survival (OS). Functionally, the activation of non-canonical WNT/PCP signaling by WNT5A (and not canonical WNT/β-catenin signaling by WNT3A) promoted the metastatic stem-cell (metSC) like behavior (i.e. self-renewal, migration, and invasion) of HGSC cells. The pharmacological inhibition of casein kinase 1 (CK1) as well as genetic ablation (dishevelled 3 knock out) of the pathway blocked the WNT5A-induced effect. Additionally, WNT/PCP pathway components were differentially expressed between healthy and tumor tissue as well as between the primary tumor and metastases. Additionally, ascites which activated WNT/PCP signaling contained the typical WNT/PCP ligand WNT5A and interestingly, patients with high levels of WNT5A protein in their ascites exhibited poor progression-free survival (PFS) and OS in comparison to patients with low or undetectable ascitic WNT5A. Together, our results suggest the existence of a positive feedback loop between tumor cells producing WNT ligands and ascites that distribute WNT activity to cancer cells in the peritoneum, in order to promote their pro-metastatic features and drive HGSC progression. Conclusions: Our results highlight the role of WNT/PCP signaling in ovarian cancerogenesis, indicate a possible therapeutic potential of CK1 inhibitors for HGSC, and strongly suggest that the detection of WNT pathway inducing activity ascites (or WNT5A levels in ascites as a surrogate marker) could be a novel prognostic tool for HGSC patients.

• MeSH
• ascites metabolismus   patologie   MeSH
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• míra přežití MeSH
• nádorové biomarkery metabolismus   MeSH
• nádorové buněčné linie MeSH
• nádorové mikroprostředí fyziologie   MeSH
• nádory vaječníků metabolismus   mortalita   patologie   MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• signální dráha Wnt * MeSH
• stupeň nádoru MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Chronická lymfocytární leukémie (CLL) je nejčastější leukémie dospělých v České republice. Výsledky z našich laboratoří ukazují, že jednou z rozhodujících molekulárních drah spojených s patogenezí CLL je nekanonická Wnt/PCP signalizace. Naše výsledky ukázaly, že klíčové komponenty Wnt/PCP dráhy jsou u CLL výrazně zvýšeny a dokáží oddělit jednotlivé podskupiny CLL lišící se klinickým průběhem a biologií onemocnění. Analýza komponent Wnt/PCP dráhy tak má potenciál (i) definovat nové diagnostické markery, (ii) předpovídat odpověď/terapeutický přínos nových terapií a (iii) být použita jako užitečný nástroj pro sledování odpovědi na léčbu a CLL relapsu. V navrhovaném projektu ověříme tyto předpoklady na vzorcích CLL dobře charakterizovaných z klinického i molekulárně-biologického hlediska. Projekt je zaměřen na studium leukemogeneze s dlouhodobým cílem zlepšit management CLL s využitím nových léčebných postupů. Splnění těchto cílů povede ke snížení nákladů na léčbu CLL díky cílenější terapii a inovativní diagnostice.; Chronic lymphocytic leukemia (CLL) is the most common leukemia among adults in the Czech Republic. The recent data from our laboratories suggest that one of the crucial molecular pathways associated with pathogenesis of CLL is Wnt/PCP signaling pathway. Our results showed that the key components of Wnt/PCP pathway are upregulated in CLL and can distinguish individual subgroups of CLL differing in clinical course and disease biology. These results suggest that components of non-canonical Wnt signaling have the potential (i) to become novel diagnostic markers, (ii) to predict a response/patient benefit from the novel therapies and (iii) to be used as a useful tool for monitoring treatment response and CLL relapse. We propose to validate these assumptions in CLL samples well characterized from clinical and molecular biology point of view. The project focuses on the study of leukemogenesis with the long term goal to improve CLL treatment by novel therapies. In summary, this can lead to the cost reduction of CLL treatment by more focused therapy and innovative diagnostics.

• MeSH
• chronická lymfatická leukemie terapie   MeSH
• membránové proteiny MeSH
• nádorové biomarkery MeSH
• nádorové mikroprostředí MeSH
• polarita buněk MeSH
• proteiny Wnt MeSH
• signální dráha Wnt MeSH

• Konspekt
• Patologie. Klinická medicína
• NLK Obory
• onkologie
• biochemie
• NLK Publikační typ
• závěrečné zprávy o řešení grantu AZV MZ ČR

BACKGROUND: Autosomal-dominant mutations in the Park8 gene encoding Leucine-rich repeat kinase 2 (LRRK2) have been identified to cause up to 40% of the genetic forms of Parkinson's disease. However, the function and molecular pathways regulated by LRRK2 are largely unknown. It has been shown that LRRK2 serves as a scaffold during activation of WNT/β-catenin signaling via its interaction with the β-catenin destruction complex, DVL1-3 and LRP6. In this study, we examine whether LRRK2 also interacts with signaling components of the WNT/Planar Cell Polarity (WNT/PCP) pathway, which controls the maturation of substantia nigra dopaminergic neurons, the main cell type lost in Parkinson's disease patients. METHODS: Co-immunoprecipitation and tandem mass spectrometry was performed in a mouse substantia nigra cell line (SN4741) and human HEK293T cell line in order to identify novel LRRK2 binding partners. Inhibition of the WNT/β-catenin reporter, TOPFlash, was used as a read-out of WNT/PCP pathway activation. The capacity of LRRK2 to regulate WNT/PCP signaling in vivo was tested in Xenopus laevis' early development. RESULTS: Our proteomic analysis identified that LRRK2 interacts with proteins involved in WNT/PCP signaling such as the PDZ domain-containing protein GIPC1 and Integrin-linked kinase (ILK) in dopaminergic cells in vitro and in the mouse ventral midbrain in vivo. Moreover, co-immunoprecipitation analysis revealed that LRRK2 binds to two core components of the WNT/PCP signaling pathway, PRICKLE1 and CELSR1, as well as to FLOTILLIN-2 and CULLIN-3, which regulate WNT secretion and inhibit WNT/β-catenin signaling, respectively. We also found that PRICKLE1 and LRRK2 localize in signalosomes and act as dual regulators of WNT/PCP and β-catenin signaling. Accordingly, analysis of the function of LRRK2 in vivo, in X. laevis revelaed that LRKK2 not only inhibits WNT/β-catenin pathway, but induces a classical WNT/PCP phenotype in vivo. CONCLUSIONS: Our study shows for the first time that LRRK2 activates the WNT/PCP signaling pathway through its interaction to multiple WNT/PCP components. We suggest that LRRK2 regulates the balance between WNT/β-catenin and WNT/PCP signaling, depending on the binding partners. Since this balance is crucial for homeostasis of midbrain dopaminergic neurons, we hypothesize that its alteration may contribute to the pathophysiology of Parkinson's disease.

• MeSH
• beta-katenin metabolismus   MeSH
• dopaminergní neurony metabolismus   MeSH
• kadheriny metabolismus   MeSH
• lidé MeSH
• LRRK2 genetika   metabolismus   MeSH
• mutace genetika   MeSH
• polarita buněk fyziologie   MeSH
• protein-serin-threoninkinasy metabolismus   MeSH
• proteomika metody   MeSH
• signální dráha Wnt fyziologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

Colorectal cancer (CRC) is the fourth leading cause of cancer mortality worldwide. Aberrant activation of WNT/β-catenin signaling present in the vast majority of CRC cases is indispensable for CRC initiation and progression, and thus is a promising target for CRC therapeutics. Hispolon is a fungal-derived polyphenol with a pronounced anticancer effect. Several hispolon derivatives, including dehydroxyhispolon methyl ether (DHME), have been chemically synthesized for developing lead molecules with stronger anticancer activity. Herein, a DHME-elicited anti-CRC effect with the underlying mechanism is reported for the first time. Specifically, DHME was found to be more cytotoxic than hispolon against a panel of human CRC cell lines, while exerting limited toxicity to normal human colon cell line CCD 841 CoN. Additionally, the cytotoxic effect of DHME appeared to rely on inducing apoptosis. This notion was evidenced by DHME-elicited upregulation of poly (ADP-ribose) polymerase (PARP) cleavage and a cell population positively stained by annexin V, alongside the downregulation of antiapoptotic B-cell lymphoma 2 (BCL-2), whereas the blockade of apoptosis by the pan-caspase inhibitor z-VAD-fmk attenuated DHME-induced cytotoxicity. Further mechanistic inquiry revealed the inhibitory action of DHME on β-catenin-mediated, T-cell factor (TCF)-dependent transcription activity, suggesting that DHME thwarted the aberrantly active WNT/β-catenin signaling in CRC cells. Notably, ectopic expression of a dominant-active β-catenin mutant (∆N90-β-catenin) abolished DHME-induced apoptosis while also restoring BCL-2 expression. Collectively, we identified DHME as a selective proapoptotic agent against CRC cells, exerting more potent cytotoxicity than hispolon, and provoking CRC cell apoptosis via suppression of the WNT/β-catenin signaling axis.

• MeSH
• apoptóza * MeSH
• Basidiomycota chemie   MeSH
• HCT116 buňky MeSH
• kolorektální nádory farmakoterapie   metabolismus   patofyziologie   MeSH
• lidé MeSH
• nádorové buněčné linie MeSH
• poly(ADP-ribosa)polymerasy metabolismus   MeSH
• protinádorové látky farmakologie   MeSH
• protoonkogenní proteiny c-bcl-2 genetika   MeSH
• regulace genové exprese u nádorů MeSH
• signální dráha Wnt účinky léků   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH