RESUMEN
In September 2023 members of the cell adhesion and cell migration research community came together to share their latest research and consider how our work might be translated for clinical practice. Alongside invited speakers, selected speakers and poster presentations, the meeting also included a round table discussion of how we might overcome the challenges associated with research translation. This meeting report seeks to highlight the key outcomes of that discussion and spark interest in the cell adhesions and cell migration research community to cross the perceived valley of death and translate our work into therapeutic benefit.
Asunto(s)
Adhesión Celular , Movimiento Celular , Animales , Humanos , Neoplasias/patología , Neoplasias/terapia , Investigación Biomédica TraslacionalRESUMEN
Prostate cancer is the second most common male cancer worldwide showing the highest rates of incidence in Western Europe. Although the measurement of serum prostate-specific antigen levels is the current gold standard in PCa diagnosis, PSA-based screening is not considered a reliable diagnosis and prognosis tool due to its lower sensitivity and poor predictive score which lead to a 22%-43% overdiagnosis, unnecessary biopsies, and over-treatment. These major limitations along with the heterogeneous nature of the disease have made PCa a very unappreciative subject for diagnostics, resulting in poor patient management; thus, it urges to identify and validate new reliable PCa biomarkers that can provide accurate information in regard to disease diagnosis and prognosis. Researchers have explored the analysis of microRNAs (miRNAs), messenger RNAs (mRNAs), small proteins, genomic rearrangements, and gene expression in body fluids and non-solid tissues in search of lesser invasive yet efficient PCa biomarkers. Although the presence of miRNAs in body fluids like blood, urine, and saliva initially sparked great interest among the scientific community; their potential use as liquid biopsy biomarkers in PCa is still at a very nascent stage with respect to other well-established diagnostics and prognosis tools. Up to date, numerous studies have been conducted in search of PCa miRNA-based biomarkers in whole blood or blood serum; however, only a few studies have investigated their presence in urine samples of which less than two tens involve the detection of miRNAs in extracellular vesicles isolated from urine. In addition, there exists some discrepancy around the identification of miRNAs in PCa urine samples due to the diversity of the urine fractions that can be targeted for analysis such as urine circulating cells, cell-free fractions, and exosomes. In this review, we aim to discuss research output from the most recent studies involving the analysis of urinary EVs for the identification of miRNA-based PCa-specific biomarkers.
RESUMEN
RhoA-mediated regulation of myosin-II activity in the actin cortex controls the ability of cells to contract and bleb during a variety of cellular processes, including cell migration and division. Cell contraction and blebbing also frequently occur as part of the cytopathic effect seen during many different viral infections. We now demonstrate that the vaccinia virus protein F11, which localizes to the plasma membrane, is required for ROCK-mediated cell contraction from 2 hr post infection. Curiously, F11-induced cell contraction is dependent on RhoC and not RhoA signaling to ROCK. Moreover, RhoC-driven cell contraction depends on the upstream inhibition of RhoD signaling by F11. This inhibition prevents RhoD from regulating its downstream effector Pak6, alleviating the suppression of RhoC by the kinase. Our observations with vaccinia have now demonstrated that RhoD recruits Pak6 to the plasma membrane to antagonize RhoC signaling during cell contraction and blebbing.
Asunto(s)
Movimiento Celular/fisiología , Quinasas p21 Activadas/metabolismo , Proteínas de Unión al GTP rho/metabolismo , Actinas/metabolismo , Línea Celular Tumoral , Humanos , Transducción de Señal , Proteína de Unión al GTP rhoA/metabolismo , Proteína rhoC de Unión a GTPRESUMEN
Members of the ezrin-radixin-moesin (ERM) family of proteins are involved in multiple aspects of cell migration by acting both as crosslinkers between the membrane, receptors and the actin cytoskeleton, and as regulators of signalling molecules that are implicated in cell adhesion, cell polarity and migration. Increasing evidence suggests that the regulation of cell signalling and the cytoskeleton by ERM proteins is crucial during cancer progression. Thus, both their expression levels and subcellular localisation would affect tumour progression. High expression of ERM proteins has been shown in a variety of cancers. Mislocalisation of ERM proteins reduces the ability of cells to form cell-cell contacts and, therefore, promotes an invasive phenotype. Similarly, mislocalisation of ERM proteins impairs the formation of receptor complexes and alters the transmission of signals in response to growth factors, thereby facilitating tumour progression. In this Commentary, we address the structure, function and regulation of ERM proteins under normal physiological conditions as well as in cancer progression, with particular emphasis on cancers of epithelial origin, such as those from breast, lung and prostate. We also discuss any recent developments that have added to the understanding of the underlying molecular mechanisms and signalling pathways these proteins are involved in during cancer progression.
Asunto(s)
Proteínas del Citoesqueleto/metabolismo , Progresión de la Enfermedad , Proteínas de la Membrana/metabolismo , Proteínas de Microfilamentos/metabolismo , Neoplasias/metabolismo , Neoplasias/patología , Proteínas del Citoesqueleto/química , Humanos , Proteínas de la Membrana/química , Proteínas de Microfilamentos/química , Modelos BiológicosRESUMEN
Cancer cells interact with endothelial cells during the process of metastatic spreading. Here, we use a small interfering RNA screen targeting Rho GTPases in cancer cells to identify Cdc42 as a critical regulator of cancer cell-endothelial cell interactions and transendothelial migration. We find that Cdc42 regulates ß1 integrin expression at the transcriptional level via the transcription factor serum response factor (SRF). ß1 integrin is the main target for Cdc42-mediating interaction of cancer cells with endothelial cells and the underlying extracellular matrix, as exogenous ß1 integrin expression was sufficient to rescue the Cdc42-silencing phenotype. We show that Cdc42 was required in vivo for cancer cell spreading and protrusion extension along blood vessels and retention in the lungs. Interestingly, transient Cdc42 depletion was sufficient to decrease experimental lung metastases, which suggests that its role in endothelial attachment is important for metastasis. By identifying ß1 integrin as a transcriptional target of Cdc42, our results provide new insight into Cdc42 function.
Asunto(s)
Integrina beta1/metabolismo , Migración Transendotelial y Transepitelial , Proteína de Unión al GTP cdc42/metabolismo , Animales , Adhesión Celular , Línea Celular Tumoral , Células Cultivadas , Células Endoteliales/citología , Células Endoteliales/metabolismo , Fibronectinas/metabolismo , Regulación Neoplásica de la Expresión Génica , Humanos , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/patología , Ratones , Ratones SCID , Metástasis de la Neoplasia , Trasplante de Neoplasias , Transporte de Proteínas , Factor de Respuesta Sérica/metabolismo , Transcripción Genética , Migración Transendotelial y Transepitelial/genética , Proteína de Unión al GTP cdc42/deficiencia , Proteína de Unión al GTP rac1/metabolismo , Proteína de Unión al GTP rhoA/metabolismoRESUMEN
The ERM proteins ezrin, radixin and moesin are adaptor proteins that link plasma membrane receptors to the actin cytoskeleton. Ezrin and moesin have been implicated in cell polarization and cell migration, but little is known about the involvement of radixin in these processes. Here we show that radixin is required for migration of PC3 prostate cancer cells, and that radixin, but not ezrin or moesin, depletion by RNA interference increases cell spread area and cell-cell adhesion mediated by adherens junctions. Radixin depletion also alters actin organization, and distribution of active phosphorylated ezrin and moesin. Similar effects were observed in MDA-MB-231 breast cancer cells. The phenotype of radixin-depleted cells is similar to that induced by constitutively active Rac1, and Rac1 is required for the radixin knockdown phenotype. Radixin depletion also increases the activity of Rac1 but not Cdc42 or RhoA. Analysis of Rac guanine nucleotide exchange factors (GEFs) suggests that radixin affects the activity of Vav GEFs. Indeed, Vav GEF depletion reverses the phenotype of radixin knockdown and reduces the effect of radixin knockdown on Rac1 activity. Our results indicate that radixin plays an important role in promoting cell migration by regulating Rac1-mediated epithelial polarity and formation of adherens junctions through Vav GEFs.
Asunto(s)
Movimiento Celular/fisiología , Proteínas del Citoesqueleto/fisiología , Proteínas de la Membrana/fisiología , Proteína de Unión al GTP rac1/fisiología , Uniones Adherentes/fisiología , Antígenos CD/metabolismo , Cadherinas/metabolismo , Adhesión Celular/fisiología , Línea Celular Tumoral , Proteínas del Citoesqueleto/genética , Proteínas del Citoesqueleto/metabolismo , Humanos , Masculino , Proteínas de la Membrana/genética , Proteínas de la Membrana/metabolismo , Neoplasias de la Próstata/genética , Neoplasias de la Próstata/metabolismo , Neoplasias de la Próstata/patología , ARN Interferente Pequeño/administración & dosificación , ARN Interferente Pequeño/genética , beta Catenina/metabolismo , Proteína de Unión al GTP rac1/genética , Proteína de Unión al GTP rac1/metabolismo , Proteína de Unión al GTP rhoA/genética , Proteína de Unión al GTP rhoA/metabolismoRESUMEN
Prostate-derived sterile 20-like kinases (PSKs) 1-α, 1-ß, and 2 are members of the germinal-center kinase-like sterile 20 family of kinases. Previous work has shown that PSK 1-α binds and stabilizes microtubules whereas PSK2 destabilizes microtubules. Here, we have investigated the activation and autophosphorylation of endogenous PSKs and show that their catalytic activity increases as cells accumulate in G(2)/M and declines as cells exit mitosis. PSKs are stimulated in synchronous HeLa cells as they progress through mitosis, and these proteins are activated catalytically during each stage of mitosis. During prophase and metaphase activated PSKs are located in the cytoplasm and at the spindle poles, and during telophase and cytokinesis stimulated PSKs are present in trans-Golgi compartments. In addition, small interfering RNA (siRNA) knockdown of PSK1-α/ß or PSK2 expression inhibits mitotic cell rounding as well as spindle positioning and centralization. These results show that PSK catalytic activity increases during mitosis and suggest that these proteins can contribute functionally to mitotic cell rounding and spindle centralization during cell division.
Asunto(s)
División Celular/fisiología , Microtúbulos/metabolismo , Mitosis/fisiología , Proteínas Quinasas/metabolismo , Huso Acromático/metabolismo , Red trans-Golgi/metabolismo , Activación Enzimática/fisiología , Células HeLa , Humanos , Microtúbulos/genética , Proteínas Quinasas/genética , Huso Acromático/genética , Red trans-Golgi/genéticaAsunto(s)
Factores de Ribosilacion-ADP/metabolismo , Factores de Intercambio de Guanina Nucleótido/metabolismo , Animales , Neoplasias de la Mama/metabolismo , Neoplasias de la Mama/patología , Receptores ErbB/metabolismo , Humanos , Modelos Biológicos , Invasividad Neoplásica , Metástasis de la Neoplasia , Transducción de SeñalRESUMEN
RhoA signaling plays a critical role in many cellular processes, including cell migration. Here we show that the vaccinia F11L protein interacts directly with RhoA, inhibiting its signaling by blocking the interaction with its downstream effectors Rho-associated kinase (ROCK) and mDia. RNA interference-mediated depletion of F11L during infection resulted in an absence of vaccinia-induced cell motility and inhibition of viral morphogenesis. Disruption of the RhoA binding site in F11L, which resembles that of ROCK, led to an identical phenotype. Thus, inhibition of RhoA signaling is required for both vaccinia morphogenesis and virus-induced cell motility.
Asunto(s)
Movimiento Celular , Transducción de Señal , Virus Vaccinia/fisiología , Proteínas Virales/metabolismo , Proteína de Unión al GTP rhoA/metabolismo , Amidas/farmacología , Animales , Línea Celular , Proteínas del Citoesqueleto , Inhibidores Enzimáticos/farmacología , Genes Virales , Células HeLa , Humanos , Péptidos y Proteínas de Señalización Intracelular , Morfogénesis , Fosfoproteínas/metabolismo , Unión Proteica , Proteínas Serina-Treonina Quinasas/metabolismo , Estructura Terciaria de Proteína , Piridinas/farmacología , Interferencia de ARN , ARN Interferente Pequeño , Proteínas Recombinantes de Fusión/metabolismo , Transfección , Virus Vaccinia/genética , Virus Vaccinia/crecimiento & desarrollo , Proteínas Virales/química , Proteínas Virales/genética , Ensamble de Virus , Quinasas Asociadas a rhoRESUMEN
Lamellipodial protrusion is regulated by Ena/VASP proteins. We identified Lamellipodin (Lpd) as an Ena/VASP binding protein. Both proteins colocalize at the tips of lamellipodia and filopodia. Lpd is recruited to EPEC and Vaccinia, pathogens that exploit the actin cytoskeleton for their own motility. Lpd contains a PH domain that binds specifically to PI(3,4)P2, an asymmetrically localized signal in chemotactic cells. Lpd's PH domain can localize to ruffles in PDGF-treated fibroblasts. Lpd overexpression increases lamellipodial protrusion velocity, an effect observed when Ena/VASP proteins are overexpressed or artificially targeted to the plasma membrane. Conversely, knockdown of Lpd expression impairs lamellipodia formation, reduces velocity of residual lamellipodial protrusion, and decreases F-actin content. These phenotypes are more severe than loss of Ena/VASP, suggesting that Lpd regulates other effectors of the actin cytoskeleton in addition to Ena/VASP.
Asunto(s)
Proteínas Portadoras/metabolismo , Moléculas de Adhesión Celular/química , Moléculas de Adhesión Celular/metabolismo , Fosfoproteínas/química , Fosfoproteínas/metabolismo , Seudópodos/metabolismo , Actinas/metabolismo , Secuencia de Aminoácidos , Sitios de Unión , Proteínas Portadoras/química , Proteínas Portadoras/genética , Moléculas de Adhesión Celular/genética , Línea Celular , Corteza Cerebral/citología , Fibroblastos/efectos de los fármacos , Adhesiones Focales/metabolismo , Regulación de la Expresión Génica , Glutatión Transferasa/metabolismo , Células HeLa , Humanos , Cinética , Lentivirus/genética , Ligandos , Proteínas de la Membrana , Proteínas de Microfilamentos , Datos de Secuencia Molecular , Neuronas/química , Fosfoproteínas/genética , Factor de Crecimiento Derivado de Plaquetas/farmacología , Isoformas de Proteínas/genética , Isoformas de Proteínas/metabolismo , Estructura Terciaria de Proteína , Seudópodos/efectos de los fármacos , ARN Interferente Pequeño/metabolismo , Proteínas Recombinantes de Fusión/metabolismo , Homología de Secuencia de Aminoácido , Vaccinia/metabolismoRESUMEN
We have previously reported that actin filaments are involved in protein transport from the Golgi complex to the endoplasmic reticulum. Herein, we examined whether myosin motors or actin comets mediate this transport. To address this issue we have used, on one hand, a combination of specific inhibitors such as 2,3-butanedione monoxime (BDM) and 1-[5-isoquinoline sulfonyl]-2-methyl piperazine (ML7), which inhibit myosin and the phosphorylation of myosin II by the myosin light chain kinase, respectively; and a mutant of the nonmuscle myosin II regulatory light chain, which cannot be phosphorylated (MRLC2(AA)). On the other hand, actin comet tails were induced by the overexpression of phosphatidylinositol phosphate 5-kinase. Cells treated with BDM/ML7 or those that express the MRLC2(AA) mutant revealed a significant reduction in the brefeldin A (BFA)-induced fusion of Golgi enzymes with the endoplasmic reticulum (ER). This delay was not caused by an alteration in the formation of the BFA-induced tubules from the Golgi complex. In addition, the Shiga toxin fragment B transport from the Golgi complex to the ER was also altered. This impairment in the retrograde protein transport was not due to depletion of intracellular calcium stores or to the activation of Rho kinase. Neither the reassembly of the Golgi complex after BFA removal nor VSV-G transport from ER to the Golgi was altered in cells treated with BDM/ML7 or expressing MRLC2(AA). Finally, transport carriers containing Shiga toxin did not move into the cytosol at the tips of comet tails of polymerizing actin. Collectively, the results indicate that 1) myosin motors move to transport carriers from the Golgi complex to the ER along actin filaments; 2) nonmuscle myosin II mediates in this process; and 3) actin comets are not involved in retrograde transport.