Interactions between living cells and nanoparticles are extensively studied to enhance the delivery of therapeutics. Nanoparticles size, shape, stiffness, and surface charge are regarded as the main features able to control the fate of cell-nanoparticle interactions. However, the clinical translation of nanotherapies has so far been limited, and there is a need to better understand the biology of cell-nanoparticle interactions. This study investigates the role of cellular mechanosensitive components in cell-nanoparticle interactions. It is demonstrated that the genetic and pharmacologic inhibition of yes-associated protein (YAP), a key component of cancer cell mechanosensing apparatus and Hippo pathway effector, improves nanoparticle internalization in triple-negative breast cancer cells regardless of nanoparticle properties or substrate characteristics. This process occurs through YAP-dependent regulation of endocytic pathways, cell mechanics, and membrane organization. Hence, the study proposes targeting YAP may sensitize triple-negative breast cancer cells to chemotherapy and increase the selectivity of nanotherapy.

• Klíčová slova
• YAP-signaling, bio-nano interactions, cancer treatment, mechanobiology, nanoparticles,
• MeSH
• lidé MeSH
• nanočástice * MeSH
• signální proteiny YAP MeSH
• signální transdukce fyziologie   MeSH
• triple-negativní karcinom prsu * farmakoterapie   metabolismus   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• signální proteiny YAP MeSH

The yes-associated protein (YAP) and the transcriptional coactivator with PDZ-binding motif (TAZ) are transcriptional coactivators, members of the Hippo signaling pathway, which play a critical role in cell growth regulation, embryonic development, regeneration, proliferation, and cancer origin and progression. The mechanism involves the nuclear binding of the un-phosphorylated YAP/TAZ complex to release the transcriptional enhanced associate domain (TEAD) from its repressors. The active ternary complex is responsible for the aforementioned biological effects. Overexpression of YAP/TAZ has been reported in cancer stem cells and tumor resistance. The resistance involves chemotherapy, targeted therapy, and immunotherapy. This review provides an overview of YAP/TAZ pathways' role in carcinogenesis and tumor microenvironment. Potential therapeutic alternatives are also discussed.

• Klíčová slova
• Hippo signaling pathway, TEAD, YAP/TAZ, carcinogenesis, cell proliferation, chemoresistance, drug resistance, immunotherapy, neoplastic stem cells, tumor microenvironment,
• MeSH
• buněčný převod mechanických signálů MeSH
• chemorezistence MeSH
• karcinogeneze metabolismus   patologie   MeSH
• lidé MeSH
• nádorové mikroprostředí * MeSH
• signální proteiny YAP * MeSH
• transkripční koaktivátor s motivem vázajícím se na PDZ metabolismus   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH
• Názvy látek
• signální proteiny YAP * MeSH
• transkripční koaktivátor s motivem vázajícím se na PDZ MeSH

The mechanoregulated proteins YAP/TAZ are involved in the adipogenic/osteogenic switch of mesenchymal stem cells (MSCs). MSC fate decision can be unbalanced by controlling substrate mechanics, in turn altering the transmission of tension through cell cytoskeleton. MSCs have been proposed for orthopedic and reconstructive surgery applications. Thus, a tight control of their adipogenic potential is required in order to avoid their drifting towards fat tissue. Substrate mechanics has been shown to drive MSC commitment and to regulate YAP/TAZ protein shuttling and turnover. The mechanism by which YAP/TAZ co-transcriptional activity is mechanically regulated during MSC fate acquisition is still debated. Here, we design few bioengineering tools suited to disentangle the contribution of mechanical from biological stimuli to MSC adipogenesis. We demonstrate that the mechanical repression of YAP happens through its phosphorylation, is purely mediated by cell spreading downstream of substrate mechanics as dictated by dimensionality. YAP repression is sufficient to prompt MSC adipogenesis, regardless of a permissive biological environment, TEAD nuclear presence or focal adhesion stabilization. Finally, by harnessing the potential of YAP mechanical regulation, we propose a practical example of the exploitation of adipogenic transdifferentiation in tumors.

• Klíčová slova
• Adipogenesis, Biocompatible hydrogels, Cell micropatterning, Cell-matrix interaction, Mechanobiology, Mechanosensing, Mesenchymal stem cells, YAP,
• MeSH
• adaptorové proteiny signální transdukční metabolismus   MeSH
• adipogeneze * MeSH
• aktiny metabolismus   MeSH
• buněčné jádro metabolismus   MeSH
• extracelulární matrix metabolismus   MeSH
• fokální adheze metabolismus   MeSH
• fosforylace MeSH
• genetická transkripce MeSH
• lidé MeSH
• nádorové buněčné linie MeSH
• pohyb buněk * MeSH
• přeprogramování buněk MeSH
• proliferace buněk MeSH
• signální proteiny YAP MeSH
• transkripční faktory metabolismus   MeSH
• tuková tkáň cytologie   MeSH
• tukové buňky metabolismus   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• adaptorové proteiny signální transdukční MeSH
• aktiny MeSH
• signální proteiny YAP MeSH
• transkripční faktory MeSH
• YAP1 protein, human MeSH Prohlížeč

Binding of the transcriptional co-activator YAP with the transcription factor TEAD stimulates growth of the heart and other organs. YAP overexpression potently stimulates fetal cardiomyocyte (CM) proliferation, but YAP's mitogenic potency declines postnatally. While investigating factors that limit YAP's postnatal mitogenic activity, we found that the CM-enriched TEAD1 binding protein VGLL4 inhibits CM proliferation by inhibiting TEAD1-YAP interaction and by targeting TEAD1 for degradation. Importantly, VGLL4 acetylation at lysine 225 negatively regulated its binding to TEAD1. This developmentally regulated acetylation event critically governs postnatal heart growth, since overexpression of an acetylation-refractory VGLL4 mutant enhanced TEAD1 degradation, limited neonatal CM proliferation, and caused CM necrosis. Our study defines an acetylation-mediated, VGLL4-dependent switch that regulates TEAD stability and YAP-TEAD activity. These insights may improve targeted modulation of TEAD-YAP activity in applications from cardiac regeneration to cancer.

• Klíčová slova
• Hippo-YAP pathway, TEAD1, VGLL4, acetylation, cardiac, cardiomyocyte, degradation, necrosis, proliferation,
• MeSH
• acetylace MeSH
• adaptorové proteiny signální transdukční metabolismus   MeSH
• DNA vazebné proteiny metabolismus   MeSH
• fosfoproteiny metabolismus   MeSH
• lidé MeSH
• novorozená zvířata MeSH
• potkani Wistar MeSH
• proliferace buněk MeSH
• protein-serin-threoninkinasy metabolismus   MeSH
• proteinové domény MeSH
• proteiny buněčného cyklu MeSH
• sekvence aminokyselin MeSH
• signální dráha Hippo MeSH
• signální proteiny YAP MeSH
• signální transdukce * MeSH
• srdce růst a vývoj   MeSH
• srdeční selhání metabolismus   patologie   MeSH
• stabilita proteinů MeSH
• stárnutí metabolismus   MeSH
• transkripční faktory TEA domény MeSH
• transkripční faktory chemie   metabolismus   MeSH
• vazba proteinů MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• adaptorové proteiny signální transdukční MeSH
• DNA vazebné proteiny MeSH
• fosfoproteiny MeSH
• protein-serin-threoninkinasy MeSH
• proteiny buněčného cyklu MeSH
• signální proteiny YAP MeSH
• Tead1 protein, mouse MeSH Prohlížeč
• transkripční faktory TEA domény MeSH
• transkripční faktory MeSH
• VGLL4 protein, mouse MeSH Prohlížeč
• Yap1 protein, mouse MeSH Prohlížeč

The tight regulation of cytoskeleton dynamics is required for a number of cellular processes, including migration, division and differentiation. YAP-TEAD respond to cell-cell interaction and to substrate mechanics and, among their downstream effects, prompt focal adhesion (FA) gene transcription, thus contributing to FA-cytoskeleton stability. This activity is key to the definition of adult cell mechanical properties and function. Its regulation and role in pluripotent stem cells are poorly understood. Human PSCs display a sustained basal YAP-driven transcriptional activity despite they grow in very dense colonies, indicating these cells are insensitive to contact inhibition. PSC inability to perceive cell-cell interactions can be restored by tampering with Tankyrase enzyme, thus favouring AMOT inhibition of YAP function. YAP-TEAD complex is promptly inactivated when germ layers are specified, and this event is needed to adjust PSC mechanical properties in response to physiological substrate stiffness. By providing evidence that YAP-TEAD1 complex targets key genes encoding for proteins involved in cytoskeleton dynamics, we suggest that substrate mechanics can direct PSC specification by influencing cytoskeleton arrangement and intracellular tension. We propose an aberrant activation of YAP-TEAD1 axis alters PSC potency by inhibiting cytoskeleton dynamics, thus paralyzing the changes in shape requested for the acquisition of the given phenotype.

• MeSH
• adaptorové proteiny signální transdukční MeSH
• angiomotiny metabolismus   MeSH
• buněčná diferenciace MeSH
• buněčné linie MeSH
• cytoskelet metabolismus   MeSH
• lidé MeSH
• lidské embryonální kmenové buňky metabolismus   MeSH
• mezoderm metabolismus   MeSH
• signální proteiny YAP genetika   metabolismus   MeSH
• signální transdukce MeSH
• transkripční faktory TEA domény genetika   metabolismus   MeSH
• vazba proteinů MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• adaptorové proteiny signální transdukční MeSH
• angiomotiny MeSH
• signální proteiny YAP MeSH
• TEAD1 protein, human MeSH Prohlížeč
• transkripční faktory TEA domény MeSH
• YAP1 protein, human MeSH Prohlížeč

Pemphigus Vulgaris (PV) is a life-threatening autoimmune disease manifested with blisters in the skin and mucosa and caused by autoantibodies against adhesion protein desmoglein-3 (Dsg3) expressed in epithelial membrane linings of these tissues. Despite many studies, the pathogenesis of PV remains incompletely understood. Recently we have shown Dsg3 plays a role in regulating the yes-associated protein (YAP), a co-transcription factor and mechanical sensor, and constraining reactive oxygen species (ROS). This study investigated the effect of PV sera as well as the anti-Dsg3 antibody AK23 on these molecules. We detected elevated YAP steady-state protein levels in PV cells surrounding blisters and perilesional regions and in keratinocytes treated with PV sera and AK23 with concomitant transient ROS overproduction. Cells treated with hydrogen peroxide also exhibited augmented nuclear YAP accompanied by reduction of Dsg3 and α-catenin, a negative regulator of YAP. As expected, transfection of α-catenin-GFP plasmid rendered YAP export from the nucleus evoked by hydrogen peroxide. In addition, suppression of total YAP was observed in hydrogen peroxide treated cells exposed to antioxidants with enhanced cell-cell adhesion being confirmed by decreased fragmentation in the dispase assay compared to hydrogen peroxide treatment alone. On the other hand, the expression of exogenous YAP disrupted intercellular junction assembly. In contrast, YAP depletion resulted in an inverse effect with augmented expression of junction assembly proteins, including Dsg3 and α-catenin capable of abolishing the effect of AK23 on Dsg3 expression. Finally, inhibition of other kinase pathways, including p38MAPK, also demonstrated suppression of YAP induced by hydrogen peroxide. Furthermore, antioxidant treatment of keratinocytes suppressed PV sera-induced total YAP accumulation. In conclusion, this study suggests that oxidative stress coupled with YAP dysregulation attributes to PV blistering, implying antioxidants may be beneficial in the treatment of PV.

• Klíčová slova
• cell-cell adhesion, keratinocyte, oxidative stress, pemphigus vulgaris, reactive oxygen species, yes-associated protein,
• MeSH
• adaptorové proteiny signální transdukční genetika   metabolismus   MeSH
• alfa-katenin metabolismus   MeSH
• antioxidancia farmakologie   terapeutické užití   MeSH
• autoprotilátky krev   imunologie   metabolismus   MeSH
• buněčná adheze účinky léků   imunologie   MeSH
• buněčné linie MeSH
• desmoglein 3 imunologie   metabolismus   MeSH
• genový knockdown MeSH
• keratinocyty MeSH
• lidé MeSH
• MAP kinasový signální systém účinky léků   imunologie   MeSH
• oxidační stres účinky léků   imunologie   MeSH
• pemfigus krev   farmakoterapie   imunologie   patologie   MeSH
• reaktivní formy kyslíku metabolismus   MeSH
• signální proteiny YAP MeSH
• studie případů a kontrol MeSH
• transkripční faktory genetika   metabolismus   MeSH
• ústní sliznice imunologie   patologie   MeSH
• zdraví dobrovolníci pro lékařské studie MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• pozorovací studie MeSH
• práce podpořená grantem MeSH
• Názvy látek
• adaptorové proteiny signální transdukční MeSH
• alfa-katenin MeSH
• antioxidancia MeSH
• autoprotilátky MeSH
• desmoglein 3 MeSH
• DSG3 protein, human MeSH Prohlížeč
• reaktivní formy kyslíku MeSH
• signální proteiny YAP MeSH
• transkripční faktory MeSH
• YAP1 protein, human MeSH Prohlížeč

The Hippo pathway effector, Yes-associated protein (YAP), is a transcriptional coactivator implicated in cholangiocarcinoma (CCA) pathogenesis. YAP is known to be regulated by a serine/threonine kinase relay module (MST1/2-LATS1/2) culminating in phosphorylation of YAP at Serine 127 and cytoplasmic sequestration. However, YAP also undergoes tyrosine phosphorylation, and the role of tyrosine phosphorylation in YAP regulation remains unclear. Herein, YAP regulation by tyrosine phosphorylation was examined in human and mouse CCA cells, as well as patient-derived xenograft (PDX) models. YAP was phosphorylated on tyrosine 357 (Y357) in CCA cell lines and PDX models. SRC family kinase (SFK) inhibition with dasatinib resulted in loss of YAPY357 phosphorylation, promoted its translocation from the nucleus to the cytoplasm, and reduced YAP target gene expression, including cell lines expressing a LATS1/2-resistant YAP mutant in which all serine residues were mutated to alanine. Consistent with these observations, precluding YAPY357 phosphorylation by site-directed mutagenesis (YAPY357F) excluded YAP from the nucleus. Targeted siRNA experiments identified LCK as the SFK that most potently mediated YAPY357 phosphorylation. Likewise, inducible CRISPR/Cas9-targeted LCK deletion decreased YAPY357 phosphorylation and its nuclear localization. The importance of LCK in CCA biology was demonstrated by clinical observations suggesting LCK expression levels were associated with early tumor recurrence following resection of CCA. Finally, dasatinib displayed therapeutic efficacy in PDX models. Mol Cancer Res; 16(10); 1556-67. ©2018 AACR.

• MeSH
• adaptorové proteiny signální transdukční genetika   MeSH
• buněčné jádro účinky léků   MeSH
• cholangiokarcinom farmakoterapie   genetika   patologie   MeSH
• cytoplazma účinky léků   MeSH
• dasatinib aplikace a dávkování   MeSH
• fosfoproteiny genetika   MeSH
• fosforylace účinky léků   MeSH
• lidé MeSH
• myši MeSH
• nádorové buněčné linie MeSH
• proliferace buněk účinky léků   MeSH
• protein-serin-threoninkinasy genetika   MeSH
• regulace genové exprese u nádorů účinky léků   MeSH
• signální proteiny YAP MeSH
• signální transdukce účinky léků   MeSH
• skupina kinas odvozených od src-genu antagonisté a inhibitory   genetika   MeSH
• transkripční faktory MeSH
• tyrosin genetika   MeSH
• tyrosinkinasa p56(lck), specifická pro lymfocyty genetika   MeSH
• xenogenní modely - testy protinádorové aktivity MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, N.I.H., Extramural MeSH
• Research Support, U.S. Gov't, Non-P.H.S. MeSH
• Názvy látek
• adaptorové proteiny signální transdukční MeSH
• dasatinib MeSH
• fosfoproteiny MeSH
• LATS1 protein, human MeSH Prohlížeč
• LCK protein, human MeSH Prohlížeč
• protein-serin-threoninkinasy MeSH
• signální proteiny YAP MeSH
• skupina kinas odvozených od src-genu MeSH
• transkripční faktory MeSH
• tyrosin MeSH
• tyrosinkinasa p56(lck), specifická pro lymfocyty MeSH
• YAP1 protein, human MeSH Prohlížeč

Phosphorylation-dependent YAP translocation is a well-known intracellular mechanism of the Hippo pathway; however, the molecular effectors governing YAP cytoplasmic translocation remains undefined. Recent findings indicate that oncogenic YAP paradoxically suppresses Wnt activity. Here, we show that Wnt scaffolding protein Dishevelled (DVL) is responsible for cytosolic translocation of phosphorylated YAP. Mutational inactivation of the nuclear export signal embedded in DVL leads to nuclear YAP retention, with an increase in TEAD transcriptional activity. DVL is also required for YAP subcellular localization induced by E-cadherin, α-catenin, or AMPK activation. Importantly, the nuclear-cytoplasmic trafficking is dependent on the p53-Lats2 or LKB1-AMPK tumor suppressor axes, which determine YAP phosphorylation status. In vivo and clinical data support that the loss of p53 or LKB1 relieves DVL-linked reciprocal inhibition between the Wnt and nuclear YAP activity. Our observations provide mechanistic insights into controlled proliferation coupled with epithelial polarity during development and human cancer.

• MeSH
• adaptorové proteiny signální transdukční metabolismus   MeSH
• aktivní transport - buněčné jádro * MeSH
• alfa-katenin metabolismus   MeSH
• buněčné jádro metabolismus   MeSH
• buňky A549 MeSH
• cytoplazma metabolismus   MeSH
• fosfoproteiny metabolismus   MeSH
• fosforylace MeSH
• HCT116 buňky MeSH
• HEK293 buňky MeSH
• kadheriny metabolismus   MeSH
• kinasy AMP aktivovaných proteinkinas MeSH
• lidé MeSH
• MFC-7 buňky MeSH
• mutace MeSH
• mutační analýza DNA MeSH
• myši nahé MeSH
• myši MeSH
• nádorový supresorový protein p53 metabolismus   MeSH
• protein dishevelled metabolismus   MeSH
• protein Wnt1 metabolismus   MeSH
• protein-serin-threoninkinasy metabolismus   MeSH
• proteinkinasy aktivované AMP metabolismus   MeSH
• proteiny buněčného cyklu MeSH
• proteiny Wnt metabolismus   MeSH
• signální dráha Hippo MeSH
• signální proteiny YAP MeSH
• transkripční faktory MeSH
• transport proteinů MeSH
• tumor supresorové geny * MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• adaptorové proteiny signální transdukční MeSH
• alfa-katenin MeSH
• DVL1 protein, human MeSH Prohlížeč
• fosfoproteiny MeSH
• kadheriny MeSH
• kinasy AMP aktivovaných proteinkinas MeSH
• nádorový supresorový protein p53 MeSH
• protein dishevelled MeSH
• protein Wnt1 MeSH
• protein-serin-threoninkinasy MeSH
• proteinkinasy aktivované AMP MeSH
• proteiny buněčného cyklu MeSH
• proteiny Wnt MeSH
• signální proteiny YAP MeSH
• STK11 protein, human MeSH Prohlížeč
• TP53 protein, human MeSH Prohlížeč
• transkripční faktory MeSH
• WNT1 protein, human MeSH Prohlížeč
• YAP1 protein, human MeSH Prohlížeč
• Yap1 protein, mouse MeSH Prohlížeč

Recent studies undoubtedly show that the mammalian target of rapamycin (mTOR) and the Hippo-Yes-associated protein 1 (YAP) pathways are important mediators of mechanical cues. The crosstalk between these pathways as well as de-regulation of their signaling has been implicated in multiple tumor types, including liver tumors. Additionally, physical cues from 3D microenvironments have been identified to alter gene expression and differentiation of different cell lineages. However, it remains incompletely understood how physical constraints originated in 3D cultures affect cell plasticity and what the key mediators are of such process. In this work, we use collagen scaffolds as a model of a soft 3D microenvironment to alter cellular size and study the mechanotransduction that regulates that process. We show that the YAP-mTOR axis is a downstream effector of 3D cellular culture-driven mechanotransduction. Indeed, we found that cell mechanics, dictated by the physical constraints of 3D collagen scaffolds, profoundly affect cellular proliferation in a YAP-mTOR-mediated manner. Functionally, the YAP-mTOR connection is key to mediate cell plasticity in hepatic tumor cell lines. These findings expand the role of YAP-mTOR-driven mechanotransduction to the control hepatic tumor cellular responses under physical constraints in 3D cultures. We suggest a tentative mechanism, which coordinates signaling rewiring with cytoplasmic restructuring during cell growth in 3D microenvironments.

• Klíčová slova
• 3D cultures, YAP, autophagy, cell plasticity, cytoskeleton, mTOR, mechanotransduction,
• Publikační typ
• časopisecké články MeSH

Deregulated Hippo pathway signaling is associated with aberrant activation of the downstream effector yes-associated protein (YAP), an emerging key oncogenic mediator in cholangiocarcinoma (CCA). In our prior work, we have demonstrated that biliary transduction of YAP along with Akt as a permissive factor induces CCA in mice. To further delineate the mechanisms associated with YAP-associated biliary oncogenesis, we have established seven malignant murine cell lines from our YAP-driven murine CCA model. These cells express the CCA markers SRY (Sex Determining Region Y)-Box 9 (SOX9), cytokeratin (CK)-7 and 19 but lack hepatocyte nuclear factor 4 alpha and alpha-smooth muscle actin, markers of hepatocellular carcinoma and cancer-associated fibroblasts, respectively. Notably, the murine CCA cells can be readily implanted into mouse livers with resultant orthotopic tumor formation. In this unique syngeneic orthotopic murine model, tumors exhibit histopathologic features resembling human CCA. We analyzed transcriptome data from YAP-associated parent CCA tumor nodules and identified a gene expression pattern associated with chromosomal instability, known as CIN25. Similarly, mate-pair sequencing of the murine CCA cells revealed chromosomal missegregation with gains and losses of several whole chromosomes demonstrating aneuploidy. Of the CIN25 genes, forkhead box M1 (Foxm1), a key cell cycle regulator, was the most significantly upregulated CIN25 gene product. Accordingly, small interfering RNA (siRNA)-mediated silencing of YAP as well as FOXM1 inhibition with thiostrepton induced CCA cell death. These preclinical data imply a role for YAP-mediated chromosomal instability in cholangiocarcinoma, and suggest FOXM1 inhibition as a therapeutic target for CCA.

• Klíčová slova
• FOXM1, SB cell lines, chromosomal instability, mate-pair sequencing,
• Publikační typ
• časopisecké články MeSH