Protein Ezrin Research Articles

Clinical and molecular features of neurofibromatosis types 1 and 2: a review of the literature

Neurofibromatosis is a neurocutaneous syndrome characterized by the development of tumors of the central or peripheral nervous system including the brain , spinal cord , organs , skin , and bones . There are three types of neurofibromatosis : type 1 (96 of cases ), type 2 (3 % of cases ), and schwannomatosis ( less than 1 % of cases ). The NF1 gene is located on chromosome 17q11.2, which encodes for a tumor suppressor protein , neurofibromin , that functions as a negative regulator of Ras / MAPK ( mitogen-activated protein kinase ) and PI3K ( phosphoinositide 3- kinases ) / mTOR ( mammalian target of rapamycin ) signaling pathways . The NF2 gene is identified on chromosome 22q12, which encodes for merlin , a tumor suppressor protein related to the proteins ezrin , radixin and moesin that modulates the activity of PI3K/AKT, Raf /MEK/ERK, and mTOR signaling pathways . In contrast , molecular insights on the different forms of schwannomatosis remain unclear . Inactivating mutations in the tumor suppressor genes MARCB1 and LZTR1 are considered responsible for a majority of cases . Recently , treatment strategies to target specific genetic or molecular events involved in their tumorigenesis are developed . This study discusses molecular pathways and related targeted therapies for neurofibromatosis type 1, type 2, and schwannomatosis and reviews recent clinical trials which involve neurofibromatosis patients . The aim of the study is to present the features and pathophysiology of neurofibromatosis , as well as modern diagnostic and therapeutic strategies related to this pathology .

Abstract PR013: Novel mechanisms of ezrin function in osteosarcoma metastasis

Abstract Ezrin, a member of the ezrin/radixin/moesin protein family, functions as a cytoskeletal linker to mediate cell adhesion, motility, proliferation, and programmed death in different tissues. Ezrin plays a critical role in osteosarcoma (OS) metastasis to the lungs. Even though ezrin has no function in proliferation of primary tumor cells, OS cells lacking ezrin do not metastasize to the lungs. Even the OS cells that extravasate in lung capillaries fail to grow metastatic foci in the absence of ezrin. The protein is believed to be in an inactive (closed) conformational state in the cytoplasm. Phosphorylated ezrin acquired an active (open) conformation and relocates to the plasma membrane. The exact molecular mechanism of how ezrin in the membrane contributes to OS metastasis remains elusive. In this study, we propose an alternative mechanism for ezrin in the cytoplasm through direct interaction with mRNA and new protein partners that are involved in stress granules. We discovered the first in class ezrin inhibitor, NSC305787, though a surface plasmon resonance screening experiment and validated its biological activities. We demonstrated that NSC305787 directly binds to ezrin protein and inhibits ezrin mediated OS metastasis in three different mouse models including a transgenic mouse with spontaneous lung metastasis and primary xenografts tumors that were removed by amputation for metastasis development. We then used the small molecule inhibitor ezrin as molecular probe to discover novel interaction partners of ezrin in the cytoplasm. Affinity column capture of ezrin and NSC305787 competition followed by mass spec analysis identified a number of novel ezrin binding proteins whose interaction was disrupted by NSC305787 in OS cell lysates. At least seven of these novel interactors were RNA binding proteins (DDX3, Caprin-1, FAK, FXR1, PABP1, EWS, FUS) that were implicated in translation regulation and found in stress granules, which are membrane-less cytoplasmic granules where particular mRNA species are stored during stress. Supporting ezrin’s role in stress granule formation, we demonstrated that ezrin knockdown or inhibition by NSC305787 affects the expression levels of a set of stress response genes. In addition to interacting with RNA binding proteins, we also identified several mRNAs preferentially associated with ezrin in OS cells by RIP-Seq (Ribonuclear protein Immuno-precipitation). Expression of ezrin binding target mRNAs correlated with presence of ezrin and can be modulated by the ezrin inhibitor. Therefore, we propose that ezrin may regulate target gene expression by directly binding to mRNA and interacting with a number of stress granule proteins. Our data challenges the conventional molecular mechanism in the field that required ezrin phosphorylation and membrane localization and proposes a novel mechanism of action in the cytoplasm. Citation Format: Aykut Uren, Emre Deniz, Taryn Shaw, Jeffrey Toretsky, Luiz O. Penalva. Novel mechanisms of ezrin function in osteosarcoma metastasis [abstract]. In: Proceedings of the AACR Special Conference: Sarcomas; 2022 May 9-12; Montreal, QC, Canada. Philadelphia (PA): AACR; Clin Cancer Res 2022;28(18_Suppl):Abstract nr PR013.

Intestinal brush border formation requires a TMIGD1-based intermicrovillar adhesion complex.

Intestinal epithelial cells absorb nutrients through the brush border, composed of dense arrays of highly ordered microvilli at their apical membranes. A protocadherin-based intermicrovillar adhesion complex localized at microvilli tips mediates microvilli packing and organization. Here, we identified a second adhesion complex localized at the proximal base region of microvilli. This complex contained the immunoglobulin superfamily member TMIGD1, which directly interacted with the microvillar scaffolding proteins EBP50 and E3KARP. Complex formation with EBP50 required the activation of EBP50 by the actin-binding protein ezrin and was enhanced by the dephosphorylation of Ser162 in the PDZ2 domain of EBP50 by the phosphatase PP1α. Binding of the EBP50-ezrin complex to TMIGD1 enhanced the dynamic turnover of EBP50 at microvilli. Enterocyte-specific inactivation of Tmigd1 in mice resulted in microvillar blebbing, loss of intermicrovillar adhesion, and perturbed brush border formation. Thus, we identified a second adhesion complex in microvilli and propose a mechanism that promotes microvillar formation and dynamics.

Docking and molecular dynamics studies of human ezrin protein with a modelled SARS-CoV-2 endodomain and their interaction with potential invasion inhibitors

Human ezrin protein interacts with SARS-CoV S endodomain and restricts virus fusion, entry, and early events of infection. In general, their binding strength and their structural stability determines their successful entry into the host cells. However, the binding affinity of these two endodomains with the ezrin protein has been elusive due to a paucity of knowledge on the 3D structure. This study modelled the endodomains of both SARS-CoV-1 and SARS-CoV-2 and then docked these models with human ezrin protein. This study establishes that the modelled endodomains of both SARS-CoV-1 and SARS-Cov-2 consisted of three disulphide bridges for self-stabilization. Protein-protein docking listed four salt bridges with a higher buried surface area between ezrin-SARS-CoV-1 endodomain compared to that of ezrin-SARS-CoV-2 with six salt bridges with lower buried surface area. Molecular simulation of the ezrin-SARS-CoV-1 endodomain showed better structural stability with lower Root Mean Square Deviation score compared to that of ezrin-SARS-CoV-2 endodomain due to the substitution of alanine with cysteine residue. Protein-ligand docking studies confirmed better ezrin-drug interaction for quercetin, minocycline, calcifediol, calcitriol, selamectin, ivermectin and ergocalciferol. However, protein–ligand simulation confirmed strong drug-protein interaction during simulation for all the above-listed drugs except for ergocalciferol which could not establish its interaction with the protein during simulation. Strong drug binding within the active site pocket therefore restricts the interaction of viral endodomain and simultaneously stabilizes the ezrin protein. Furthermore, the higher stability between the ezrin after their interaction with the drug moiety could restrict the virus fusion and the infection. This study provides a basis for further development of these drug molecules to clinical trials aiming to identify potential drug molecules which can treat COVID-19 infection.

The membrane-actin linker ezrin acts as a sliding anchor.

Protein linkages to filamentous (F)–actin provide the cell membrane with mechanical stability and support intricate membrane architectures. However, the actin cytoskeleton is highly dynamic and undergoes rapid changes in shape during cell motility and other processes. The molecular mechanisms that generate a mechanically robust yet fluid connection between the membrane and actin cytoskeleton remain poorly understood. Here, we adapted a single-molecule optical trap assay to examine how the prototypical membrane-actin linker ezrin acts to anchor F-actin to the cell membrane. We find that ezrin forms a complex that slides along F-actin over micrometer distances while resisting detachment by forces oriented perpendicular to the filament axis. The ubiquity of ezrin and analogous proteins suggests that sliding anchors such as ezrin may constitute an important but overlooked element in the construction of the actin cytoskeleton.

Ezrin and CD44 participate in the internalization process of Coxiella burnetii into non-phagocytic cells.

Ezrin protein is involved in the interaction of actin cytoskeleton with membrane receptors such as CD44. It regulates plasma membrane dynamics and intracellular signaling. Coxiella burnetii, the etiologic agent of Q fever, is internalized into host cell through a poorly characterized molecular mechanism. Here we analyzed the role of ezrin and CD44 in the C. burnetii internalization by HeLa cells. The knockdown of ezrin and CD44 inhibited the bacterial uptake. Interestingly, at early stages of C. burnetii internalization, ezrin was recruited to the cell membrane fraction and phosphorylated. Moreover, the overexpression of non-phosphorylatable and phosphomimetic ezrin mutants decreased and increased the bacterial entry, respectively. A decrease in the internalization of C. burnetii was observed by the overexpression of CD44 truncated forms containing the intracellular or the extracellular domains. Interestingly, the CD44 mutant was unable to interact with ERM proteins decreased the bacterial internalization. These findings demonstrate the participation of ezrin in the internalization process of C. burnetii in non-phagocytic cells. Additionally, we present evidence that CD44 receptor would be involved in that process.

Induction of Autophagy Promotes Clearance of RHOP23H Aggregates and Protects From Retinal Degeneration.

Autophagy is a critical metabolic process that acts as a major self-digestion and recycling pathway contributing to maintain cellular homeostasis. An emerging field of research supports the therapeutic modulation of autophagy for treating human neurodegenerative disorders, in which toxic aggregates are accumulated in neurons. Our previous study identified Ezrin protein as an inhibitor of autophagy and lysosomal functions in the retina; thus, in turn, identifying it as a potential pharmacological target for increasing retinal cell clearance to treat inherited retinal dystrophies in which misfolded proteins have accumulated. This study aimed to verify the therapeutic inhibition of Ezrin to induce clearance of toxic aggregates in a mouse model for a dominant form of retinitis pigmentosa (i.e., RHOP23H/+). We found that daily inhibition of Ezrin significantly decreased the accumulation of misfolded RHOP23H aggregates. Remarkably, induction of autophagy, by a drug-mediated pulsatile inhibition of Ezrin, promoted the lysosomal clearance of disease-linked RHOP23H aggregates. This was accompanied with a reduction of endoplasmic reticulum (ER)-stress, robust decrease of photoreceptors' cell death, amelioration in both retinal morphology and function culminating in a better preservation of vision. Our study opens new perspectives for a pulsatile pharmacological induction of autophagy as a mutation-independent therapy paving the way toward a more effective therapeutic strategy to treat these devastating retinal disorders due to an accumulation of intracellular toxic aggregates.

Amygdalin Exerts Antitumor Activity in Taxane-Resistant Prostate Cancer Cells

Simple SummaryThe natural compound amygdalin is popular among tumor patients as an alternative treatment option. However, knowledge about its precise mode of action remains poor. In the present study, amygdalin is shown to inhibit growth and disseminative properties of taxane-resistant prostate cancer cells. Further investigation is warranted to determine the role of amygdalin in the setting of metastasized prostate cancer.Despite recent advances in the treatment of metastatic prostate cancer (PCa), resistance development after taxane treatments is inevitable, necessitating effective options to combat drug resistance. Previous studies indicated antitumoral properties of the natural compound amygdalin. However, whether amygdalin acts on drug-resistant tumor cells remains questionable. An in vitro study was performed to investigate the influence of amygdalin (10 mg/mL) on the growth of a panel of therapy-naïve and docetaxel- or cabazitaxel-resistant PCa cell lines (PC3, DU145, and LNCaP cells). Tumor growth, proliferation, clonal growth, and cell cycle progression were investigated. The cell cycle regulating proteins (phospho)cdk1, (phospho)cdk2, cyclin A, cyclin B, p21, and p27 and the mammalian target of rapamycin (mTOR) pathway proteins (phospho)Akt, (phospho)Raptor, and (phospho)Rictor as well as integrin β1 and the cytoskeletal proteins vimentin, ezrin, talin, and cytokeratin 8/18 were assessed. Furthermore, chemotactic activity and adhesion to extracellular matrix components were analyzed. Amygdalin dose-dependently inhibited tumor growth and reduced tumor clones in all (parental and resistant) PCa cell lines, accompanied by a G0/G1 phase accumulation. Cell cycle regulating proteins were significantly altered by amygdalin. A moderate influence of amygdalin on tumor cell adhesion and chemotaxis was observed as well, paralleled by modifications of cytoskeletal proteins and the integrin β1 expression level. Amygdalin may, therefore, block tumor growth and disseminative characteristics of taxane-resistant PCa cells. Further studies are warranted to determine amygdalin’s value as an antitumor drug.

Preprint Highlight: Interphase microtubule disassembly is a signaling cue that drives cell rounding at mitotic entry.

Metaphase cell rounding is an important step during cell division; however, the cell cycle regulation of the ezrin, radixin, and moesin (ERM) proteins which drive cell rounding is not well understood. Using a small molecule screen, the authors unexpectedly found that microtubule disassembly activates ERM proteins and cell rounding. The authors determined that microtubule disassembly acts as a cell cycle signal to activate ERM proteins via GEF-H1 and the kinase SLK. This new model suggests that as cells transition from interphase to metaphase, the observed change in microtubule dynamics is not only important for spindle assembly but also helps drive cortical actomyosin remodeling to enable cell rounding.

Myosin 18A is a novel checkpoint regulator in B cell differentiation and antibody-mediated immunity

Abstract The B cell antibody response is tightly regulated to facilitate pathogen-specific immunity and prevent self-reactivity, with actin cytoskeleton dynamics playing an important role in regulating B cell activation. The newly discovered unconventional myosin family protein Myosin 18A (Myo18A) is most closely related to Myo2A, and regulates important cellular processes in non-lymphoid cells. Myo18A is expressed in both precursor and mature B cells, and interacts with ezrin, Myo2A and tyrosine phosphorylated proteins suggesting that it may regulate physiological functions of B cells. We investigated the function of Myo18A in antibody-mediated immunity by generating B cell-conditional Myo18A-deficient mice. Basally, Myo18A deficiency led to expansion of both bone marrow progenitor B cells, and mature B cells in secondary lymphoid organs. Myo18A-deficient mice displayed serum IgM hyperglobulinemia with greater levels of splenic IgM secreting cells, with older mice switching to IgG1 hyperglobulinemia and autoantibody development. Immunization of Myo18A-deficient mice with inactivated influenza virus led to development of more potent neutralizing antibodies against the major antigen hemagglutinin, associated with early expansion and persistent accumulation of antigen-specific germinal center B cells. In vitro stimulation with TLR7 and BCR ligands revealed a greater ability of Myo18A-deficient B cells to differentiate into antibody secreting cells, and induce Blimp-1 expression. Overall, our study demonstrates that Myo18A is a novel negative regulator of B cell homeostasis, differentiation, self-tolerance and humoral immunity. Supported by an NIH grant (R21 AI117350) to Neetu Gupta

The prognostic value of actin-binding proteins fascin and ezrin in patients with squamous cell carcinoma of the head and neck

Rationale: During neoplastic transformation, epithelial cells become mobile, which is one of the main mechanisms of metastatic disease and recurrence. Cell motility is regulated by actin-binding proteins, which ensure the association/dissociation of actin filaments and their interaction with the cell membrane. Previously, we have shown the presence of actin-binding proteins in the serum from patients with squamous cell carcinoma of the head and neck (HNSCC); however, their association with the development of metastases and relapses in cancer patients has not been sufficiently studied.
 Aim: To evaluate the serum levels of actin-binding proteins fascin-1 and ezrin in patients with HNSCC depending on the disease recurrence and lymphatic metastasis.
 Materials and methods: Serum fascin-1 and ezrin levels before combination therapy were measured with ELISA assay in 30 HNSCC (T1-4N0-2M0) patients (mean age 56 7 years).
 Results: The median fascin-1 level was significantly higher in the patients with lymphatic metastases, compared to those without metastases: 0.64 (0.40; 5.89) vs 6.35 (1.72; 8.35) ng/mL, respectively (p 0.001). At 12 to 36 months after combination therapy, the disease relapsed in 12 (40%) patients. Ezrin levels were significantly higher in the relapsed patients, compared to those without a relapse within 3 years after combination therapy: 2.55 (2.35; 2.75) vs 1.93 (1.87; 2.5) ng/mL (p = 0.02). The ROC analysis showed an association between fascin-1 serum levels with metastatic disease (AUC = 0.71, 95% confidence interval 0.570.85) and an association between ezrin levels and the disease relapse (AUC = 0.76, 95% confidence interval 0.570.94).
 Conclusion: These indicators can be used for the development of minimally invasive early detection of metastases in lymphatic nodes and for the prognosis of HNSCC recurrence.

BifA Triggers Phosphorylation of Ezrin to Benefit Streptococcus equi subsp. zooepidemicus Survival from Neutrophils Killing.

Streptococcus equi subsp. zooepidemicus (SEZ) ATCC35246 can invade the brain and cause severe neutrophils infiltration in brain tissue. This microorganism can survive and reproduce to an extremely high CFU burden (108–109/organ) under stressful neutrophils infiltration circumstances. The aim of this research is to explore the mechanism of the SEZ hypervirulent strain with its specific bifA gene which avoids being eliminated by neutrophils in the brain. We isolated the primary mouse neutrophils to treat SEZ WT and bifA gene defective (ΔBif) strains. The ΔBif strain had a weakened function of defending against neutrophils killing in vitro. The interaction between BifA and ezrin proteins in neutrophils were identified by co-IP and immunoblot. In neutrophils, the BifA interacts with ezrin and triggers the phosphorylation of ezrin at its Thr567 site in a PKC-dependent manner, then the excessive elevation of phosphorylated-ezrin recruits Dbl and activates Rac1. Since the Rac1 is closely relevant to several critical cellular functions, its abnormal activation will lead to neutrophils dysfunction and benefit to SEZ survival from neutrophils killing. Our findings reveal a novel consequence of BifA and ERM family protein (for ezrin, radixin, moesin) interaction, which happens between BifA and ezrin in neutrophils and contributes to SEZ survival in the brain. BifA should be considered as a potential target for drug development to prevent SEZ infection.

Nrf2 Activation Protects Against Organic Dust and Hydrogen Sulfide Exposure Induced Epithelial Barrier Loss and K. pneumoniae Invasion.

Agriculture workers report various respiratory symptoms owing to occupational exposure to organic dust (OD) and various gases. Previously, we demonstrated that pre-exposure to hydrogen sulfide (H2S) alters the host response to OD and induces oxidative stress. Nrf2 is a master-regulator of host antioxidant response and exposures to toxicants is known to reduce Nrf2 activity. The OD exposure-induced lung inflammation is known to increase susceptibility to a secondary microbial infection. We tested the hypothesis that repeated exposure to OD or H2S leads to loss of Nrf2, loss of epithelial cell integrity and that activation of Nrf2 rescues this epithelial barrier dysfunction. Primary normal human bronchial epithelial (NHBE) cells or mouse precision cut-lung slices (PCLS) were treated with media, swine confinement facility organic dust extract (ODE) or H2S or ODE+H2S for one or five days. Cells were also pretreated with vehicle control (DMSO) or RTA-408, a Nrf2 activator. Acute exposure to H2S and ODE+H2S altered the cell morphology, decreased the viability as per the MTT assay, and reduced the Nrf2 expression as well as increased the keap1 levels in NHBE cells. Repeated exposure to ODE or H2S or ODE+H2S induced oxidative stress and cytokine production, decreased tight junction protein occludin and cytoskeletal protein ezrin expression, disrupted epithelial integrity and resulted in increased Klebsiella pneumoniae invasion. RTA-408 (pharmacological activator of Nrf2) activated Nrf2 by decreasing keap1 levels and reduced ODE+H2S-induced changes including reversing loss of barrier integrity, inflammatory cytokine production and microbial invasion in PCLS but not in NHBE cell model. We conclude that Nrf2 activation has a partial protective function against ODE and H2S.

Piezo1 activation attenuates thrombin-induced blebbing in breast cancer cells.

ABSTRACTCancer cells exploit a variety of migration modes to leave primary tumors and establish metastases, including amoeboid cell migration, which is typically reliant on bleb formation. Here we demonstrate that thrombin induces dynamic blebbing in the MDA-MB-231 breast cancer cell line and confirm that protease-activated receptor 1 (PAR1) activation is sufficient to induce this effect. Cell confinement has been implicated as a driving force in bleb-based migration. Unexpectedly, we found that gentle contact compression, exerted using a custom built ‘cell press’ to mechanically stimulate cells, reduced thrombin-induced blebbing. Thrombin-induced blebbing was similarly attenuated using the small molecule Yoda1, an agonist of the mechanosensitive Ca2+ channel Piezo1, and this attenuation was impaired in Piezo1-depleted cells. Additionally, Piezo1 activation suppressed thrombin-induced phosphorylation of ezrin, radixin and moesin (ERM) proteins, which are implicated in the blebbing process. Our results provide mechanistic insights into Piezo1 activation as a suppressor of dynamic blebbing, specifically that which is induced by thrombin.

Oocyte ERM and EWI Proteins Are Involved in Mouse Fertilization.

In mammalian fertilization, the link between the oocyte plasma membrane and underneath cytoskeleton has often been associated to key elements of successful gamete fusion, like microvilli shaping or CD9 function, but its effective role has poorly been studied. EWI-2 and EWI-F as cis partners of CD9, and ERM proteins (Ezrin, Radixin and Moesin) that both attach to the actin cytoskeleton and to the EWI are part of the molecules that make the link between the oocyte membrane and its cytoskeleton. This study aims to assay through siRNA inhibition, the involvement of these ERM and EWI molecules in mouse fertilization, their role in the microvilli morphology of the egg but also their possible contribution to the cortical tension, a parameter that reflects the mechanical behavior of the oocyte cortex. Whereas inhibiting separately the expression of each protein had no effect on fertilization, the combined inhibition of either EWI-2/EWI-F or the three ERM triggered a significant decrease of the fertilization index. This inhibition seems to correlate with an increase in the radius of curvature of the oocyte microvilli. It also causes a decrease of the oocyte cortical tension. These results show the importance of EWI-2 and EWI–F and ERM proteins in the smooth running of a fertilization event and support their involvement in the microvilli architecture of the oocyte and in its mechanical properties.

Expression Patterns of Ezrin and AJAP1 and Clinical Significance in Breast Cancer.

Ezrin and adherens junction-associated protein 1 (AJAP1) are structural proteins which are involved in numerous human malignancies. However, little is known about the relationship between them in breast cancer. This study was set out to investigate the relationship between them and to further explore the mechanism of AJAP1-mediating cytoskeleton in breast cancer progression. Ezrin and AJAP1 expressions were detected in 377 samples of breast cancer by immunohistochemistry, and different expression patterns between AJAP1 and Ezrin with clinicopathological parameters were analyzed. Besides, univariate and multivariate Cox models were used to evaluate their prognostic potential. Enzyme-linked immunosorbent assay, Western blot, qRT-PCR, and phalloidin staining of F-actin were used to explore the relationship and the mechanism between AJAP1 and Ezrin in cytoskeleton arrangement. 377 cases of breast cancer results showed that AJAP1 expression was negatively related with histological grade and lymph node involvement and could be an independent prognosis marker of breast cancer. AJAP1 expression tended to be higher in the Ezrin-negative expression case. Patients with AJAP1negative and Ezrinpositive expression had a worse prognosis (p < 0.0001) and shorter DFS (p = 0.015). More importantly, AJAP1 depletion increased the cell ability of F-actin formation through promoting Ezrin expression. AJAP1 depletion might mediate breast cancer malignancy potential through promoting Ezrin expression and cytoskeleton formation.

Could Moesin Be a New Marker for Indicating Progression in Endometrial Cancer?

AimThis study aims to determine an important parameter in progression from pre-invasive lesions of endometrium to endometrial cancer and also evaluate the effect of this parameter on the progression of endometrial cancer.Material and MethodIn our study,30 patients with normal endometrial tissue (group 1), 56 patients who had endometrial hyperplasia without atypia (group 2), 36 patients who had endometrial hyperplasia with atypia (group 3), and 63 patients with endometrial cancer (group 4) were included. Age, parity, body-mass index, systemic diseases, and tumor markers of patients were evaluated. Expression levels of Ezrin, Radixin, and Moesin proteins were immunohistochemically evaluated in terms of frequency, intensity, and score value.ResultsWhen we compared hyperplasia cases with or without atypia; frequency, and score value of ezrin expression and frequency, intensity, and score value of moesin expression was significantly higher in patients who had hyperplasia with atypia (p:0.000 p:0.001 p:0.003, p:0.032 p: 0.035 p:0.015 p:0.005, respectively). It was observed that the frequency and score value of moesin expression were significantly higher in patients with endometrial cancer when compared with patients who had hyperplasia with atypia (p:0.003 p:0.045). The frequency of moesin expression was significantly higher in patients who had postoperative mortality (p:0.030 p:0.039).ConclusionIncreased frequency of moesin expression in the preoperative period in patients with atypical hyperplasia should alert the surgeon in terms of malignancy. If the frequency of moesin expression increases in cases with endometrial cancer, the patient should be followed closely in terms of progression in the postoperative period.

Disruption of amphetamine sensitization by alteration of dendritic thin spines in the nucleus accumbens core.

Repeated injections of psychomotor stimulants like amphetamine (AMPH) to rodents can induce behavioral sensitization, which represents a long-lasting craving that is usually observed in human addicts. Behavioral sensitization is characteristically maintained for a long duration, accompanied by structural plasticity in some brain areas involved in reward circuitry. For example, it increased dendritic spine densities in the nucleus accumbens (NAcc), which is considered to reflect neurophysiological changes at this site, leading to addictive behaviors. The ezrin, radixin, and moesin (ERM) proteins regulate spine maturity by modifying their phosphorylation at the C-terminal region. We previously showed that ERM phosphorylation is reduced by AMPH in the NAcc core, suggesting that ERM-mediated spine changes at this site might be associated with AMPH sensitization. To test this hypothesis, we administered AMPH to rats according to a sensitization development schedule, with lentivirus encoding a phosphomimetic pseudo-active mutant of radixin (Rdx T564D) in the NAcc core, and examined dendritic spines at this site. We found that compared to acute AMPH, AMPH sensitization increased thin spine density with a similar ratio of filopodia-like to mature thin spines. However, with Rdx T564D, the density of thin spines increased, with augmented filopodia-like thin spines, resulting in no AMPH sensitization. These results indicate that Rdx T564D forces thin spines to immaturity and thereby inhibits AMPH sensitization, for which an increase in mature thin spines is normally necessary. These findings provide significant clues to our understanding of the role of dendritic spines in mediating the development of psychomotor stimulant addiction.

Expression of ezrin protein and phosphorylated ezrin in pelvic endometriotic lesions

ObjectiveTo evaluate the expression of Ezrin and Phosphorylated Ezrin (Phospho-Ezrin) in endometriosis lesions and its relation to the menstrual cycle phase, stage of endometriosis, histological classification, and clinical symptoms. Material and methodsThe authors conducted a retrospective study, with endometriotic lesions collected from women with endometriosis (n = 57) who underwent laparoscopy from 2017 to 2018. The expression of Ezrin and Phosphorylated Ezrin proteins was analyzed by immunohistochemistry. ResultsAll the endometriotic lesions contained immunostaining for Ezrin in the glands. Phosphorylated Ezrin was expressed in the stroma of all endometriotic lesions. There was no difference in the Ezrin and Phosphorylated Ezrin's expression in the retrocervical, ovarian, superficial, and intestinal lesions in the same patient. Dysmenorrhea, dyspareunia, acyclic pain, infertility, and dysuria were similar in the three groups of Ezrin staining. There was an inversely proportional relationship between severe dyschezia and Ezrin's intensity, being 66.7% of Ezrin 1 (weak intensity), 36.7 Ezrin 2 (moderate intensity), and 10.0% of Ezrin 3 (p = 0.013). Regarding Phospho-Ezrin there wasn't a significant difference between all the analyzed variables. Histological classification and menstrual cycle phase had also no significant difference between Ezrin and Phospho-Ezrin immunostaining. ConclusionEzrin protein and Phospho-Ezrin can be considered important markers to elucidate the mechanisms related to migration and attachment of endometriotic lesions. It is still unclear if Ezrin and Phospho-Ezrin are a cause or consequence of endometriosis. Further studies comparing different types of lesions and eutopic endometrium are necessary to elucidate the role of these proteins in the pathogenesis of endometriosis.

Correlations of mRNA Levels among Efflux Transporters, Transcriptional Regulators, and Scaffold Proteins in Non-Small-Cell Lung Cancer.

Multidrug resistance (MDR) due to enhanced drug efflux activity of tumor cells can severely impact the efficacy of antitumor therapies. We recently showed that increased activity of the efflux transporter P-glycoprotein (P-gp) associated with activation of Snail transcriptional regulators may be mediated mainly by moesin in lung cancer cells. Here, we aimed to systematically evaluate the relationships among mRNA expression levels of efflux transporters (P-gp, breast cancer resistance protein (BCRP), and multidrug resistance-associated protein 2 (MRP2)), scaffold proteins (ezrin (Ezr), radixin (Rdx), and moesin (Msn); ERM proteins), and SNAI family members (Snail, Slug, and Smac) in clinical lung cancer and noncancer samples. We found high correlations between relative (cancer/noncancer) mRNA expression levels of Snail and Msn, Msn and P-gp, Slug and MRP2, and Smuc and BCRP. These findings support our previous conclusion that Snail regulates P-gp activity via Msn and further suggest that Slug and Smuc may contribute to the functional regulation of MRP2 and BCRP, respectively, in lung cancer cells. This trial is registered with UMIN000023923.