162 results on '"Signal Initiation"'
Search Results
2. MicroRNAs as possible indicators of drug sensitivity in breast cancer cell lines.
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Uhr, Katharina, Prager-van der Smissen, Wendy J. C., Heine, Anouk A. J., Ozturk, Bahar, van Jaarsveld, Marijn T. M., Boersma, Antonius W. M., Jager, Agnes, Wiemer, Erik A. C., Smid, Marcel, Foekens, John A., and Martens, John W. M.
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CELL lines , *CANCER cells , *BREAST cancer , *BREAST , *CELL cycle , *DRUG resistance - Abstract
MicroRNAs (miRNAs) regulate gene expression post-transcriptionally. In this way they might influence whether a cell is sensitive or resistant to a certain drug. So far, only a limited number of relatively small scale studies comprising few cell lines and/or drugs have been performed. To obtain a broader view on miRNAs and their association with drug response, we investigated the expression levels of 411 miRNAs in relation to drug sensitivity in 36 breast cancer cell lines. For this purpose IC50 values of a drug screen involving 34 drugs were associated with miRNA expression data of the same breast cancer cell lines. Since molecular subtype of the breast cancer cell lines is considered a confounding factor in drug association studies, multivariate analysis taking subtype into account was performed on significant miRNA-drug associations which retained 13 associations. These associations consisted of 11 different miRNAs and eight different drugs (among which Paclitaxel, Docetaxel and Veliparib). The taxanes, Paclitaxel and Docetaxel, were the only drugs having miRNAs in common: hsa-miR-187-5p and hsa-miR-106a-3p indicative of drug resistance while Paclitaxel sensitivity alone associated with hsa-miR-556-5p. Tivantinib was associated with hsa-let-7d-5p and hsa-miR-18a-5p for sensitivity and hsa-miR-637 for resistance. Drug sensitivity was associated with hsa-let-7a-5p for Bortezomib, hsa-miR-135a-3p for JNJ-707 and hsa-miR-185-3p for Panobinostat. Drug resistance was associated with hsa-miR-182-5p for Veliparib and hsa-miR-629-5p for Tipifarnib. Pathway analysis for significant miRNAs was performed to reveal biological roles, aiding to find a potential mechanistic link for the observed associations with drug response. By doing so hsa-miR-187-5p was linked to the cell cycle G2-M checkpoint in line with this checkpoint being the target of taxanes. In conclusion, our study shows that miRNAs could potentially serve as biomarkers for intrinsic drug resistance and that pathway analyses can provide additional information in this context. [ABSTRACT FROM AUTHOR]
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- 2019
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3. Killing two birds with one stone: How the respiratory syncytial virus polymerase initiates transcription and replication.
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Noton, Sarah L., Tremaglio, Chadene Z., and Fearns, Rachel
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RESPIRATORY syncytial virus , *PEDIATRIC respiratory diseases , *GENETIC transcription , *VIRAL replication , *MESSENGER RNA - Abstract
The article offers information on the study on Respiratory syncytial virus as the major cause of respiratory disease in infants and young children. Topics discussed include information on the transcription and replication of the virus; discussions on the engagement of the polymerase in the either of the process, transcription and replication; and the information on the how the relative levels of Messenger RNA (mRNA), antigenome, and genome synthesis controlled.
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- 2019
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4. An enhanced password authentication scheme for session initiation protocol with perfect forward secrecy.
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Qiu, Shuming, Xu, Guoai, Ahmad, Haseeb, and Guo, Yanhui
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SESSION Initiation Protocol (Computer network protocol) , *BIOMETRIC identification , *MULTIMEDIA communications , *CRYPTOGRAPHY , *COMPUTER security - Abstract
The Session Initiation Protocol (SIP) is an extensive and esteemed communication protocol employed to regulate signaling as well as for controlling multimedia communication sessions. Recently, Kumari et al. proposed an improved smart card based authentication scheme for SIP based on Farash’s scheme. Farash claimed that his protocol is resistant against various known attacks. But, we observe some accountable flaws in Farash’s protocol. We point out that Farash’s protocol is prone to key-compromise impersonation attack and is unable to provide pre-verification in the smart card, efficient password change and perfect forward secrecy. To overcome these limitations, in this paper we present an enhanced authentication mechanism based on Kumari et al.’s scheme. We prove that the proposed protocol not only overcomes the issues in Farash’s scheme, but it can also resist against all known attacks. We also provide the security analysis of the proposed scheme with the help of widespread AVISPA (Automated Validation of Internet Security Protocols and Applications) software. At last, comparing with the earlier proposals in terms of security and efficiency, we conclude that the proposed protocol is efficient and more secure. [ABSTRACT FROM AUTHOR]
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- 2018
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5. Treatment effect of mTOR-inhibition on tissue composition of renal angiomyolipomas in tuberous sclerosis complex (TSC).
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Brakemeier, Susanne, Vogt, Lars, Adams, Lisa, Zukunft, Bianca, Diederichs, Gerd, Hamm, Bernd, Budde, Klemens, and Makowski, Marcus R.
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MTOR inhibitors , *ANGIOMYOLIPOMA , *TUBEROUS sclerosis diagnosis , *TUBEROUS sclerosis , *EVEROLIMUS , *THERAPEUTICS - Abstract
Purpose: Tuberous sclerosis complex (TSC)-associated renal angiomyolipoma (AML) have a high lifetime risk of acute bleeding. MTOR-inhibitors are a promising novel treatment for TSC-AML, however adequate response to therapy can be difficult to assess. Early changes in MRI signal may serve as a novel early indicator for a satisfactory response to mTOR-inhibitor therapy of AML. Materials and methods: Thirty-eight patients with the definite diagnosis of tuberous sclerosis receiving everolimus therapy and n = 19 patients without specific therapy were included. 1.5 Tesla MRI was performed including sequences with a selective fat suppression. Patients were investigated prior to the initiation of therapy (baseline) and after <3 months (n = 21 patients), 3 to 6 months (n = 32) and 18 to 24 months (n = 28). Signal and size changes of renal AMLs were assessed at all different timepoints. Signal-to-noise-ratio (SNR), contrast-to-noise-ratio (CNR) and size of angiomyolipomas were evaluated. Results: Signal changes in 273 AMLs were evaluated. A significant and strong decrease of the CNR of AMLs following the initiation of therapy was measured in the fat-suppressed MR sequence at all time points, compared to the baseline: From 7.41±6.98 to 3.84±6.25 (p ≤ 0.05p = 0.002), 3.36±6.93 (p<0.0001), and 2.50±6.68 (p<0.0001) after less than 3 months, 3–6 months or 18–24 months of everolimus treatment, respectively. Also, a significant, however less pronounced, reduction of angiomyolipoma size in the different groups was measured (from baseline 2022.2±2657.7 mm2 to 1854.4±1670.9 mm2 (p = 0.009), 1875.5±3190.1 mm2 (p<0.001), and 1365.8 ± 1628.8 mm2 (p<0.0001) after less than 3 months, 3–6 months or 18–24 months of everolimus treatment, respectively). No significant changes in CNR (p>0.05) and size (p>0.05) were measured in the control group. Conclusion: mTOR inhibitor therapy in TSC patients results in an early and pronounced fatty transformation of AMLs on MRI. Fatty transformation could represent a novel early indicator of response to therapy in this patient collective. [ABSTRACT FROM AUTHOR]
- Published
- 2017
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6. Modulation of corticospinal output in agonist and antagonist proximal arm muscles during motor preparation.
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Neige, Cécilia, Massé-Alarie, Hugo, Gagné, Martin, Bouyer, Laurent J., and Mercier, Catherine
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MOTOR ability , *EVOKED potentials (Electrophysiology) , *PYRAMIDAL tract , *IMMUNOMODULATORS , *MUSCLE physiology - Abstract
Previous studies have shown modulation of corticospinal output of the agonist muscle when a known-movement is prepared but withheld until a response signal appearance, reflecting motor preparation processes. However, modulation in the antagonist muscles has not been described, despite the fact that reaching movements require precise coordination between the activation of agonist and antagonist muscles. In this study, participants performed an instructed-delay reaction time (RT) task, with randomized elbow flexion and extension movements. The aim was to assess the time course modulation of corticospinal output in two antagonist muscles, by simultaneously quantified the amplitude of motor evoked potentials (MEPs) in biceps brachii and triceps brachii, and the amplitude and direction of elbow movements evoked by transcranial magnetic stimulation (TMS). Depending on the prepared movement direction, a specific modulation of corticospinal output was observed, MEPs and TMS-evoked movements amplitude being relatively greater for extension compared to flexion. At the end of motor preparation, a decrease in MEPs amplitude was observed for both biceps brachii and triceps brachii, regardless of the prepared movement direction. In contrast, the probability of evoking movement in the flexion direction and the amplitude of TMS-evoked movement decreased at the end of preparation for flexion, but not for extension. Together, these results confirm the existence of inhibitory processes at the end of the motor preparation, probably to avoid a premature motor response. Moreover, they provide evidence of differences in the corticospinal control of elbow flexor and extensor muscles with patterns of modulation that are not necessarily reciprocal during motor preparation. [ABSTRACT FROM AUTHOR]
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- 2017
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7. Interglomerular Connectivity within the Canonical and GC-D/Necklace Olfactory Subsystems.
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Uytingco, Cedric R., Puche, Adam C., and Munger, Steven D.
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CANONICAL transformations , *SMELL disorders , *CELL receptors , *GLOMERULAR filtration rate , *SEMIOCHEMICALS - Abstract
The mammalian main olfactory system contains several subsystems that differ not only in the receptors they express and the glomerular targets they innervate within the main olfactory bulb (MOB), but also in the strategies they use to process odor information. The canonical main olfactory system employs a combinatorial coding strategy that represents odorant identity as a pattern of glomerular activity. By contrast, the "GC-D/necklace" olfactory subsystem—formed by olfactory sensory neurons expressing the receptor guanylyl cyclase GC-D and their target necklace glomeruli (NGs) encircling the caudal MOB—is critical for the detection of a small number of semiochemicals that promote the acquisition of food preferences. The formation of these socially-transmitted food preferences requires the animal to integrate information about two types of olfactory stimuli: these specialized social chemosignals and the food odors themselves. However, the neural mechanisms with which the GC-D/necklace subsystem processes this information are unclear. We used stimulus-induced increases in intrinsic fluorescence signals to map functional circuitry associated with NGs and canonical glomeruli (CGs) in the MOB. As expected, CG-associated activity spread laterally through both the glomerular and external plexiform layers associated with activated glomeruli. Activation of CGs or NGs resulted in activity spread between the two types of glomeruli; there was no evidence of preferential connectivity between individual necklace glomeruli. These results support previous anatomical findings that suggest the canonical and GC-D/necklace subsystems are functionally connected and may integrate general odor and semiochemical information in the MOB. [ABSTRACT FROM AUTHOR]
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- 2016
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8. Real-time single-cell characterization of the eukaryotic transcription cycle reveals correlations between RNA initiation, elongation, and cleavage
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Simon Alamos, Donald Hansen, Hernan G. Garcia, Elizabeth Eck, Jonathan Liu, Yang Joon Kim, and Meghan A Turner
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Embryology ,Transcription, Genetic ,Statistical methods ,Cell ,Gene Expression ,Biochemistry ,chemistry.chemical_compound ,Signal Initiation ,Transcription (biology) ,RNA polymerase ,Biology (General) ,Ecology ,Hydrolysis ,Transcriptional Control ,Messenger RNA ,Drosophila Melanogaster ,Mechanisms of Signal Transduction ,Statistics ,Eukaryota ,Animal Models ,Nucleic acids ,Monte Carlo method ,Physical sciences ,Insects ,medicine.anatomical_structure ,Experimental Organism Systems ,Computational Theory and Mathematics ,Modeling and Simulation ,Drosophila ,Single-Cell Analysis ,Research Article ,Signal Transduction ,Arthropoda ,QH301-705.5 ,DNA transcription ,Computational biology ,Cleavage (embryo) ,Cellular and Molecular Neuroscience ,Model Organisms ,Gene Types ,Genetics ,medicine ,Animals ,Gene Regulation ,Molecular Biology ,Ecology, Evolution, Behavior and Systematics ,Reporter gene ,Embryos ,Eukaryotic transcription ,Organisms ,Biology and Life Sciences ,RNA ,Cell Biology ,Invertebrates ,Research and analysis methods ,chemistry ,Animal Studies ,Mathematical and statistical techniques ,RNA Cleavage ,Zoology ,Entomology ,Mathematics ,Transcription Factors ,Developmental Biology ,Reporter Genes - Abstract
The eukaryotic transcription cycle consists of three main steps: initiation, elongation, and cleavage of the nascent RNA transcript. Although each of these steps can be regulated as well as coupled with each other, their in vivo dissection has remained challenging because available experimental readouts lack sufficient spatiotemporal resolution to separate the contributions from each of these steps. Here, we describe a novel application of Bayesian inference techniques to simultaneously infer the effective parameters of the transcription cycle in real time and at the single-cell level using a two-color MS2/PP7 reporter gene and the developing fruit fly embryo as a case study. Our method enables detailed investigations into cell-to-cell variability in transcription-cycle parameters as well as single-cell correlations between these parameters. These measurements, combined with theoretical modeling, suggest a substantial variability in the elongation rate of individual RNA polymerase molecules. We further illustrate the power of this technique by uncovering a novel mechanistic connection between RNA polymerase density and nascent RNA cleavage efficiency. Thus, our approach makes it possible to shed light on the regulatory mechanisms in play during each step of the transcription cycle in individual, living cells at high spatiotemporal resolution., Author summary Live cell imaging using fluorescence microscopy provides an exciting way to visualize the transcription cycle in living organisms with great amounts of precision. However, the output of these technologies is often complex and can be hard to interpret. We have developed a computational framework for analyzing the transcription cycle that quantifies rates of RNA initiation, elongation, and cleavage, given input datasets from live cell imaging. Using the developing fruit fly embryo as a case study, we demonstrate that our methodology can quantitatively describe the whole transcription cycle at single-cell resolution. These results allow us to investigate a plethora of avenues, from couplings between different aspects of the transcription cycle at the single-cell level to comparisons with theoretical predictions of distributions of elongation rates across cells. We envision our methodology to provide a unified computational framework for the analysis of transcriptional data obtained from live cell imaging.
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- 2021
9. A Feed-Forward Circuit Linking Wingless, Fat-Dachsous Signaling, and the Warts-Hippo Pathway to Drosophila Wing Growth
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Gary Struhl and Myriam Zecca
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Cell signaling ,Life Cycles ,Signal transduction ,Epithelium ,Signal Initiation ,Larvae ,Animal Cells ,Gene expression ,Morphogenesis ,Medicine and Health Sciences ,Drosophila Proteins ,Homeostasis ,Wings, Animal ,Biology (General) ,General Neuroscience ,Drosophila Melanogaster ,Mechanisms of Signal Transduction ,Wnt signaling pathway ,Gene Expression Regulation, Developmental ,Nuclear Proteins ,Signaling cascades ,Eukaryota ,Animal Models ,Cell biology ,Insects ,Experimental Organism Systems ,Bone Morphogenetic Proteins ,DPP signaling cascade ,Drosophila ,Gene Cloning ,Cellular Types ,Anatomy ,General Agricultural and Biological Sciences ,Morphogen ,Research Article ,animal structures ,Arthropoda ,QH301-705.5 ,Protocadherin ,Wnt1 Protein ,Biology ,Research and Analysis Methods ,General Biochemistry, Genetics and Molecular Biology ,Model Organisms ,Animals ,Molecular Biology Techniques ,Molecular Biology ,Wing ,General Immunology and Microbiology ,Decapentaplegic ,Mechanism (biology) ,Parietal Cells ,Organisms ,Biology and Life Sciences ,Epithelial Cells ,Molecular Development ,Invertebrates ,Wnt Proteins ,Morphogens ,Biological Tissue ,Animal Studies ,Zoology ,Entomology ,Function (biology) ,Cloning ,Developmental Biology - Abstract
Development of the Drosophila wing—a paradigm of organ development—is governed by 2 morphogens, Decapentaplegic (Dpp, a BMP) and Wingless (Wg, a Wnt). Both proteins are produced by defined subpopulations of cells and spread outwards, forming gradients that control gene expression and cell pattern as a function of concentration. They also control growth, but how is unknown. Most studies have focused on Dpp and yielded disparate models in which cells throughout the wing grow at similar rates in response to the grade or temporal change in Dpp concentration or to the different amounts of Dpp “equalized” by molecular or mechanical feedbacks. In contrast, a model for Wg posits that growth is governed by a progressive expansion in morphogen range, via a mechanism in which a minimum threshold of Wg sustains the growth of cells within the wing and recruits surrounding “pre-wing” cells to grow and enter the wing. This mechanism depends on the capacity of Wg to fuel the autoregulation of vestigial (vg)—the selector gene that specifies the wing state—both to sustain vg expression in wing cells and by a feed-forward (FF) circuit of Fat (Ft)/Dachsous (Ds) protocadherin signaling to induce vg expression in neighboring pre-wing cells. Here, we have subjected Dpp to the same experimental tests used to elucidate the Wg model and find that it behaves indistinguishably. Hence, we posit that both morphogens act together, via a common mechanism, to control wing growth as a function of morphogen range., Drosophila wing growth depends on the progressive outward spread of the morphogens Decapentaplegic (a member of the BMP family) and Wingless (a member of the Wnt family) via their capacity to sustain the growth of wing cells and to induce neighboring cells to grow and enter the wing.
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- 2021
10. The Dishevelled, EGL-10 and Pleckstrin (DEP) Domain-Containing Protein DEPDC7 Binds to CARMA2 and CARMA3 Proteins, and Regulates NF-κB Activation.
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D′ Andrea, Egildo Luca, Ferravante, Angela, Scudiero, Ivan, Zotti, Tiziana, Reale, Carla, Pizzulo, Maddalena, De La Motte, Luigi Regenburgh, De Maio, Chiara, Mazzone, Pellegrino, Telesio, Gianluca, Vito, Pasquale, and Stilo, Romania
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GENETIC regulation , *NF-kappa B , *CELLULAR signal transduction , *TRANSCRIPTION factors , *MOLECULAR biology , *G protein coupled receptors - Abstract
The molecular complexes containing BCL10, MALT1 and CARMA proteins (CBM complex) have been recently identified as a key component in the signal transduction pathways that regulate activation of Nuclear Factor kappaB (NF-κB) transcription factor. Herein we identified the DEP domain-containing protein DEPDC7 as cellular binding partners of CARMA2 and CARMA3 proteins. DEPDC7 displays a cytosolic distribution and its expression induces NF-κB activation. Conversely, shRNA-mediated abrogation of DEPDC7 results in impaired NF-κB activation following G protein-coupled receptors stimulation, or stimuli that require CARMA2 and CARMA3, but not CARMA1. Thus, this study identifies DEPDC7 as a CARMA interacting molecule, and provides evidence that DEPDC7 may be required to specifically convey on the CBM complex signals coming from activated G protein-coupled receptors. [ABSTRACT FROM AUTHOR]
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- 2014
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11. Celastrol Stimulates Hypoxia-Inducible Factor-1 Activity in Tumor Cells by Initiating the ROS/Akt/p70S6K Signaling Pathway and Enhancing Hypoxia-Inducible Factor-1α Protein Synthesis.
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Han, Xiaoxi, Sun, Shengkun, Zhao, Ming, Cheng, Xiang, Chen, Guozhu, Lin, Song, Guan, Yifu, and Yu, Xiaodan
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HYPOXIA-inducible factor 1 , *CANCER cells , *PROTEIN synthesis , *CELLULAR signal transduction , *MEDICINAL plants , *NEOVASCULARIZATION , *OXIDATIVE stress , *PREVENTION - Abstract
Celastrol, a tripterine derived from the traditional Chinese medicine plant Tripterygium wilfordii Hook F. (“Thunder of God Vine”), has been reported to have multiple effects, such as anti-inflammation, suppression of tumor angiogenesis, inhibition of tumor growth, induction of apoptosis and protection of cells against human neurodegenerative diseases. However, the mechanisms that underlie these functions are not well defined. In this study, we reported for the first time that Celastrol could induce HIF-1α protein accumulation in multiple cancer cell lines in an oxygen-independent manner and that the enhanced HIF-1α protein entered the nucleus and promoted the transcription of the HIF-1 target genes VEGF and Glut-1. Celastrol did not influence HIF-1α transcription. Instead, Celastrol induced the accumulation of the HIF-1α protein by inducing ROS and activating Akt/p70S6K signaling to promote HIF-1α translation. In addition, we found that the activation of Akt by Celastrol was transient. With increased exposure time, inhibition of Hsp90 chaperone function by Celastrol led to the subsequent depletion of the Akt protein and thus to the suppression of Akt activity. Moreover, in HepG2 cells, the accumulation of HIF-1α increased the expression of BNIP3, which induced autophagy. However, HIF-1α and BNIP3 did not influence the cytotoxicity of Celastrol because the main mechanism by which Celastrol kills cancer cells is through stimulating ROS-mediated JNK activation and inducing apoptosis. Furthermore, our data showed that the dose required for Celastrol to induce HIF-1α protein accumulation and enhance HIF-1α transcriptional activation was below its cytotoxic threshold. A cytotoxic dose of Celastrol for cancer cells did not display cytotoxicity in LO2 normal human liver cells, which indicated that the novel functions of Celastrol in regulating HIF-1 signaling and inducing autophagy might be used in new applications, such as in anti-inflammation and protection of cells against human neurodegenerative diseases. Future studies regarding these applications are required. [ABSTRACT FROM AUTHOR]
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- 2014
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12. Prediction of severity and subtype of fibrosing disease using model informed by inflammation and extracellular matrix gene index
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Amin Cheikhi, Cecelia C. Yates, Dana R. Julian, Carol Feghali-Bostwick, Zariel I. Johnson, Yvette P. Conley, James Lyons-Weiler, and Sarah E Wheeler
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0301 basic medicine ,Cell signaling ,Biopsy ,Gene Expression ,Disease ,Signal transduction ,Bioinformatics ,Inflammatory bowel disease ,Scleroderma ,0302 clinical medicine ,Signal Initiation ,Fibrosis ,Medicine and Health Sciences ,Medicine ,Skin ,Multidisciplinary ,Chemotaxis ,Mechanisms of Signal Transduction ,Age Factors ,Signaling cascades ,cAMP signaling cascade ,Extracellular Matrix ,Cell Motility ,Intercellular Signaling Peptides and Proteins ,Chemokines ,Algorithms ,Research Article ,Cell biology ,MAPK signaling cascades ,Science ,Surgical and Invasive Medical Procedures ,Gastroenterology and Hepatology ,Extracellular Matrix Signaling ,Models, Biological ,03 medical and health sciences ,Extracellular ,Genetics ,Humans ,030203 arthritis & rheumatology ,Autoimmune disease ,Inflammation ,Scleroderma, Systemic ,business.industry ,Gene Expression Profiling ,Inflammatory Bowel Disease ,Case-control study ,Biology and Life Sciences ,Bayes Theorem ,medicine.disease ,Gene expression profiling ,030104 developmental biology ,Case-Control Studies ,business - Abstract
Fibrosis is a chronic disease with heterogeneous clinical presentation, rate of progression, and occurrence of comorbidities. Systemic sclerosis (scleroderma, SSc) is a rare rheumatic autoimmune disease that encompasses several aspects of fibrosis, including highly variable fibrotic manifestation and rate of progression. The development of effective treatments is limited by these variabilities. The fibrotic response is characterized by both chronic inflammation and extracellular remodeling. Therefore, there is a need for improved understanding of which inflammation-related genes contribute to the ongoing turnover of extracellular matrix that accompanies disease. We have developed a multi-tiered method using Naive Bayes modeling that is capable of predicting level of disease and clinical assessment of patients based on expression of a curated 60-gene panel that profiles inflammation and extracellular matrix production in the fibrotic disease state. Our novel modeling design, incorporating global and parametric-based methods, was highly accurate in distinguishing between severity groups, highlighting the importance of these genes in disease. We refined this gene set to a 12-gene index that can accurately identify SSc patient disease state subsets and informs knowledge of the central regulatory pathways in disease progression.
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- 2020
13. Data-Derived Modeling Characterizes Plasticity of MAPK Signaling in Melanoma.
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Bernardo-Faura, Marti, Massen, Stefan, Falk, Christine S., Brady, Nathan R., and Eils, Roland
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MELANOMA treatment , *CELLULAR signal transduction , *SOMATIC mutation , *MITOGEN-activated protein kinases , *CELL proliferation , *NEOVASCULARIZATION , *CANCER genetics - Abstract
The majority of melanomas have been shown to harbor somatic mutations in the RAS-RAF-MEK-MAPK and PI3K-AKT pathways, which play a major role in regulation of proliferation and survival. The prevalence of these mutations makes these kinase signal transduction pathways an attractive target for cancer therapy. However, tumors have generally shown adaptive resistance to treatment. This adaptation is achieved in melanoma through its ability to undergo neovascularization, migration and rearrangement of signaling pathways. To understand the dynamic, nonlinear behavior of signaling pathways in cancer, several computational modeling approaches have been suggested. Most of those models require that the pathway topology remains constant over the entire observation period. However, changes in topology might underlie adaptive behavior to drug treatment. To study signaling rearrangements, here we present a new approach based on Fuzzy Logic (FL) that predicts changes in network architecture over time. This adaptive modeling approach was used to investigate pathway dynamics in a newly acquired experimental dataset describing total and phosphorylated protein signaling over four days in A375 melanoma cell line exposed to different kinase inhibitors. First, a generalized strategy was established to implement a parameter-reduced FL model encoding non-linear activity of a signaling network in response to perturbation. Next, a literature-based topology was generated and parameters of the FL model were derived from the full experimental dataset. Subsequently, the temporal evolution of model performance was evaluated by leaving time-defined data points out of training. Emerging discrepancies between model predictions and experimental data at specific time points allowed the characterization of potential network rearrangement. We demonstrate that this adaptive FL modeling approach helps to enhance our mechanistic understanding of the molecular plasticity of melanoma. [ABSTRACT FROM AUTHOR]
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- 2014
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14. Expression and Characterization of Purinergic Receptors in Rat Middle Meningeal Artery–Potential Role in Migraine.
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Haanes, Kristian Agmund and Edvinsson, Lars
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PURINERGIC receptors , *MENINGEAL artery , *LABORATORY rats , *MIGRAINE , *GENE expression , *PATHOLOGICAL physiology , *CELL membranes - Abstract
The dura mater and its vasculature have for decades been central in the hypothesis of migraine and headache pathophysiology. Although recent studies have questioned the role of the vasculature as the primary cause, dural vessel physiology is still relevant in understanding the complex pathophysiology of migraine. The aim of the present study was to isolate the middle meningeal artery (MMA) from rodents and characterize their purinergic receptors using a sensitive wire myograph method and RT-PCR. The data presented herein suggest that blood flow through the MMA is, at least in part, regulated by purinergic receptors. P2X1 and P2Y6 receptors are the strongest contractile receptors and, surprisingly, ADPβS caused contraction most likely via P2Y1 or P2Y13 receptors, which is not observed in other arteries. Adenosine addition, however, caused relaxation of the MMA. The adenosine relaxation could be inhibited by SCH58261 (A2A receptor antagonist) and caffeine (adenosine receptor antagonist). This gives one putative molecular mechanism for the effect of caffeine, often used as an adjuvant remedy of cranial pain. Semi-quantitative RT-PCR expression data for the receptors correlate well with the functional findings. Together these observations could be used as targets for future understanding of the in vivo role of purinergic receptors in the MMA. [ABSTRACT FROM AUTHOR]
- Published
- 2014
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15. An Image-Based Genetic Assay Identifies Genes in T1D Susceptibility Loci Controlling Cellular Antiviral Immunity in Mouse.
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Liao, Juan, Jijon, Humberto B., Kim, Ira R., Goel, Gautam, Doan, Aivi, Sokol, Harry, Bauer, Hermann, Herrmann, Bernhard G., Lassen, Kara G., and Xavier, Ramnik J.
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TYPE 1 diabetes , *DISEASE susceptibility , *LOCUS (Genetics) , *ANTIVIRAL agents , *IMMUNITY , *LABORATORY mice - Abstract
The pathogenesis of complex diseases, such as type 1 diabetes (T1D), derives from interactions between host genetics and environmental factors. Previous studies have suggested that viral infection plays a significant role in initiation of T1D in genetically predisposed individuals. T1D susceptibility loci may therefore be enriched in previously uncharacterized genes functioning in antiviral defense pathways. To identify genes involved in antiviral immunity, we performed an image-based high-throughput genetic screen using short hairpin RNAs (shRNAs) against 161 genes within T1D susceptibility loci. RAW 264.7 cells transduced with shRNAs were infected with GFP-expressing herpes simplex virus type 1 (HSV-1) and fluorescent microscopy was performed to assess the viral infectivity by fluorescence reporter activity. Of the 14 candidates identified with high confidence, two candidates were selected for further investigation, Il27 and Tagap. Administration of recombinant IL-27 during viral infection was found to act synergistically with interferon gamma (IFN-γ) to activate expression of type I IFNs and proinflammatory cytokines, and to enhance the activities of interferon regulatory factor 3 (IRF3). Consistent with a role in antiviral immunity, Tagap-deficient macrophages demonstrated increased viral replication, reduced expression of proinflammatory chemokines and cytokines, and decreased production of IFN-β. Taken together, our unbiased loss-of-function genetic screen identifies genes that play a role in host antiviral immunity and delineates roles for IL-27 and Tagap in the production of antiviral cytokines. [ABSTRACT FROM AUTHOR]
- Published
- 2014
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16. Genomic and Genotoxic Responses to Controlled Weathered-Oil Exposures Confirm and Extend Field Studies on Impacts of the Deepwater Horizon Oil Spill on Native Killifish.
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Pilcher, Whitney, Miles, Scott, Tang, Song, Mayer, Greg, and Whitehead, Andrew
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GENETIC toxicology , *OIL pollution of water , *DEEPWATER Horizon (Drilling rig) , *OIL spills , *KILLIFISHES , *FISH ecology , *POLLUTION - Abstract
To understand the ecotoxicological impacts of the Deepwater Horizon oil spill, field studies provide a context for ecological realism but laboratory-based studies offer power for connecting biological effects with specific causes. As a complement to field studies, we characterized genome-wide gene expression responses of Gulf killifish (Fundulus grandis) to oil-contaminated waters in controlled laboratory exposures. Transcriptional responses to the highest concentrations of oiled water in the laboratory were predictive of field-observed responses that coincided with the timing and location of major oiling. The transcriptional response to the low concentration (∼10-fold lower than the high concentration) was distinct from the high concentration and was not predictive of major oiling in the field. The high concentration response was characterized by activation of the molecular signaling pathway that facilitates oil metabolism and oil toxicity. The high concentration also induced DNA damage. The low concentration invoked expression of genes that may support a compensatory response, including genes associated with regulation of transcription, cell cycle progression, RNA processing, DNA damage, and apoptosis. We conclude that the gene expression response detected in the field was a robust indicator of exposure to the toxic components of contaminating oil, that animals in the field were exposed to relatively high concentrations that are especially damaging to early life stages, and that such exposures can damage DNA. [ABSTRACT FROM AUTHOR]
- Published
- 2014
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17. Pannexin 1 Channels Play Essential Roles in Urothelial Mechanotransduction and Intercellular Signaling.
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Negoro, Hiromitsu, Urban-Maldonado, Marcia, Liou, Louis S., Spray, David C., Thi, Mia M., and Suadicani, Sylvia O.
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PANNEXINS , *TRANSITIONAL cell carcinoma , *MECHANOTRANSDUCTION (Cytology) , *CELL communication , *CELLULAR signal transduction , *BLADDER cancer , *EPITHELIAL cells - Abstract
Urothelial cells respond to bladder distension with ATP release, and ATP signaling within the bladder and from the bladder to the CNS is essential for proper bladder function. In other cell types, pannexin 1 (Panx1) channels provide a pathway for mechanically-induced ATP efflux and for ATP-induced ATP release through interaction with P2X7 receptors (P2X7Rs). We report that Panx1 and P2X7R are functionally expressed in the bladder mucosa and in immortalized human urothelial cells (TRT-HU1), and participate in urothelial ATP release and signaling. ATP release from isolated rat bladders induced by distention was reduced by the Panx1 channel blocker mefloquine (MFQ) and was blunted in mice lacking Panx1 or P2X7R expression. Hypoosmotic shock induced YoPro dye uptake was inhibited by MFQ and the P2X7R blocker A438079 in TRT-HU1 cells, and was also blunted in primary urothelial cells derived from mice lacking Panx1 or P2X7R expression. Rinsing-induced mechanical stimulation of TRT-HU1 cells triggered ATP release, which was reduced by MFQ and potentiated in low divalent cation solution (LDPBS), a condition known to enhance P2X7R activation. ATP signaling evaluated as intercellular Ca2+ wave radius was significantly larger in LDPBS, reduced by MFQ and by apyrase (ATP scavenger). These findings indicate that Panx1 participates in urothelial mechanotransduction and signaling by providing a direct pathway for mechanically-induced ATP release and by functionally interacting with P2X7Rs. [ABSTRACT FROM AUTHOR]
- Published
- 2014
- Full Text
- View/download PDF
18. The FYVE Domain of Smad Anchor for Receptor Activation (SARA) Is Required to Prevent Skin Carcinogenesis, but Not in Mouse Development.
- Author
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Chang, Huang-Ming, Lin, Yu-Ying, Tsai, Pei-Chun, Liang, Chung-Tiang, and Yan, Yu-Ting
- Subjects
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SKIN cancer prevention , *CARCINOGENESIS , *LABORATORY mice , *TRANSFORMING growth factors-beta , *CELLULAR signal transduction - Abstract
Smad Anchor for Receptor Activation (SARA) has been reported as a critical role in TGF-β signal transduction by recruiting non-activated Smad2/3 to the TGF-β receptor and ensuring appropriate subcellular localization of the activated receptor-bound complex. However, controversies still exist in previous reports. In this study, we describe the expression of two SARA isoforms, SARA1 and SARA2, in mice and report the generation and characterization of SARA mutant mice with FYVE domain deletion. SARA mutant mice developed normally and showed no gross abnormalities. Further examination showed that the TGF-β signaling pathway was indeed altered in SARA mutant mice, with the downregulation of Smad2 protein expression. The decreasing expression of Smad2 was caused by enhancing Smurf2-mediated proteasome degradation pathway. However, the internalization of TGF-β receptors into the early endosome was not affected in SARA mutant mouse embryonic fibroblasts (MEFs). Moreover, the downregulation of Smad2 in SARA mutant MEFs was not sufficient to disrupt the diverse cellular biological functions of TGF-β signaling, including growth inhibition, apoptosis, senescence, and the epithelial-to-mesenchymal transition. Our results indicate that SARA is not involved in the activation process of TGF-β signal transduction. Using a two-stage skin chemical carcinogenesis assay, we found that the loss of SARA promoted skin tumor formation and malignant progression. Our data suggest a protective role of SARA in skin carcinogenesis. [ABSTRACT FROM AUTHOR]
- Published
- 2014
- Full Text
- View/download PDF
19. Cathepsin S Signals via PAR2 and Generates a Novel Tethered Ligand Receptor Agonist.
- Author
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Elmariah, Sarina B., Reddy, Vemuri B., and Lerner, Ethan A.
- Subjects
- *
CATHEPSINS , *CELLULAR signal transduction , *LIGANDS (Biochemistry) , *PROTEASE-activated receptors , *SERINE proteinases , *PROTEIN expression , *ENZYME activation - Abstract
Protease-activated receptor-2 is widely expressed in mammalian epithelial, immune and neural tissues. Cleavage of PAR2 by serine proteases leads to self-activation of the receptor by the tethered ligand SLIGRL. The contribution of other classes of proteases to PAR activation has not been studied in detail. Cathepsin S is a widely expressed cysteine protease that is upregulated in inflammatory conditions. It has been suggested that cathepsin S activates PAR2. However, cathepsin S activation of PAR2 has not been demonstrated directly nor has the potential mechanism of activation been identified. We show that cathepsin S cleaves near the N-terminus of PAR2 to expose a novel tethered ligand, KVDGTS. The hexapeptide KVDGTS generates downstream signaling events specific to PAR2 but is weaker than SLIGRL. Mutation of the cathepsin S cleavage site prevents receptor activation by the protease while KVDGTS retains activity. In conclusion, the range of actions previously ascribed to cysteine cathepsins in general, and cathepsin S in particular, should be expanded to include molecular signaling. Such signaling may link together observations that had been attributed previously to PAR2 or cathepsin S individually. These interactions may contribute to inflammation. [ABSTRACT FROM AUTHOR]
- Published
- 2014
- Full Text
- View/download PDF
20. Fine Tuning of the UPR by the Ubiquitin Ligases Siah1/2.
- Author
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Scortegagna, Marzia, Kim, Hyungsoo, Li, Jian-Liang, Yao, Hang, Brill, Laurence M., Han, Jaeseok, Lau, Eric, Bowtell, David, Haddad, Gabriel, Kaufman, Randal J., and Ronai, Ze'ev A.
- Subjects
- *
UBIQUITIN ligases , *ENDOPLASMIC reticulum , *ISCHEMIA , *PROLINE hydroxylase , *HYDROXYLATION - Abstract
The endoplasmic reticulum (ER) responds to changes in intracellular homeostasis through activation of the unfolded protein response (UPR). Yet, it is not known how UPR-signaling coordinates adaptation versus cell death. Previous studies suggested that signaling through PERK/ATF4 is required for cell death. We show that high levels of ER stress (i.e., ischemia-like conditions) induce transcription of the ubiquitin ligases Siah1/2 through the UPR transducers PERK/ATF4 and IRE1/sXBP1. In turn, Siah1/2 attenuates proline hydroxylation of ATF4, resulting in its stabilization, thereby augmenting ER stress output. Conversely, ATF4 activation is reduced upon Siah1/2 KD in cultured cells, which attenuates ER stress-induced cell death. Notably, Siah1a+/−::Siah2−/− mice subjected to neuronal ischemia exhibited smaller infarct volume and were protected from ischemia-induced death, compared with the wild type (WT) mice. In all, Siah1/2 constitutes an obligatory fine-tuning mechanism that predisposes cells to death under severe ER stress conditions. [ABSTRACT FROM AUTHOR]
- Published
- 2014
- Full Text
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21. An In Vivo EGF Receptor Localization Screen in C. elegans Identifies the Ezrin Homolog ERM-1 as a Temporal Regulator of Signaling.
- Author
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Haag, Andrea, Gutierrez, Peter, Bühler, Alessandra, Walser, Michael, Yang, Qiutan, Langouët, Maeva, Kradolfer, David, Fröhli, Erika, Herrmann, Christina J., Hajnal, Alex, and Escobar-Restrepo, Juan M.
- Subjects
- *
EZRIN , *CAENORHABDITIS elegans genetics , *EPITHELIAL cells , *GENETIC regulation , *EPIDERMAL growth factor receptors , *HOMOLOGY (Biology) - Abstract
The subcellular localization of the epidermal growth factor receptor (EGFR) in polarized epithelial cells profoundly affects the activity of the intracellular signaling pathways activated after EGF ligand binding. Therefore, changes in EGFR localization and signaling are implicated in various human diseases, including different types of cancer. We have performed the first in vivo EGFR localization screen in an animal model by observing the expression of the EGFR ortholog LET-23 in the vulval epithelium of live C. elegans larvae. After systematically testing all genes known to produce an aberrant vulval phenotype, we have identified 81 genes regulating various aspects of EGFR localization and expression. In particular, we have found that ERM-1, the sole C. elegans Ezrin/Radixin/Moesin homolog, regulates EGFR localization and signaling in the vulval cells. ERM-1 interacts with the EGFR at the basolateral plasma membrane in a complex distinct from the previously identified LIN-2/LIN-7/LIN-10 receptor localization complex. We propose that ERM-1 binds to and sequesters basolateral LET-23 EGFR in an actin-rich inactive membrane compartment to restrict receptor mobility and signaling. In this manner, ERM-1 prevents the immediate activation of the entire pool of LET-23 EGFR and permits the generation of a long-lasting inductive signal. The regulation of receptor localization thus serves to fine-tune the temporal activation of intracellular signaling pathways. [ABSTRACT FROM AUTHOR]
- Published
- 2014
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22. Reassembly of JIP1 Scaffold Complex in JNK MAP Kinase Pathway Using Heterologous Protein Interactions.
- Author
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Moon, Jiyoung and Park, Sang-Hyun
- Subjects
- *
MITOGEN-activated protein kinases , *CELLULAR signal transduction , *PROTEIN-protein interactions , *CELL death , *CYTOLOGY , *MOLECULAR docking - Abstract
Formation of signaling protein complexes is crucial for proper signal transduction. Scaffold proteins in MAP kinase pathways are thought to facilitate complex assembly, thereby promoting efficient and specific signaling. To elucidate the assembly mechanism of scaffold complexes in mammals, we attempted to rationally rewire JIP1-dependent JNK MAP kinase pathway via alternative assembly of JIP1 complex. When JIP1-JNK docking interaction in the complex was replaced with heterologous protein interaction domains, such as PDZ domains and JNK-binding domains, a functional scaffold complex was reconstituted, and JNK signaling was rescued. Reassembly of JIP1 complex using heterologous protein interactions was sufficient for restoring of JNK MAP kinase pathway to induce signaling responses, including JNK activation and cell death. These results suggest a simple yet modular mechanism for JIP1 scaffold assembly in mammals. [ABSTRACT FROM AUTHOR]
- Published
- 2014
- Full Text
- View/download PDF
23. Alteration of Light-Dependent Gene Regulation by the Absence of the RCO-1/RCM-1 Repressor Complex in the Fungus Neurospora crassa.
- Author
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Ruger-Herreros, Carmen, Gil-Sánchez, María del Mar, Sancar, Gencer, Brunner, Michael, and Corrochano, Luis M.
- Subjects
- *
TRANSCRIPTION factors , *GENETIC regulation , *PHOTORECEPTORS , *NEUROSPORA crassa , *CELLULAR signal transduction , *GENETIC repressors , *MESSENGER RNA - Abstract
The activation of transcription by light in the fungus Neurospora crassa requires the White Collar Complex (WCC), a photoreceptor and transcription factor complex. After light reception two WCCs interact and bind the promoters of light-regulated genes to activate transcription. This process is regulated by VVD, a small photoreceptor that disrupts the interaction between WCCs and leads to a reduction in transcription after long exposures to light. The N. crassa RCO-1/RCM-1 repressor complex is the homolog of the Tup1-Ssn6 repressor complex in yeast, and its absence modifies photoadaptation. We show that the absence of the RCO-1/RCM-1 repressor complex leads to several alterations in transcription that are gene-specific: an increase in the accumulation of mRNAs in the dark, a repression of transcription, and a derepression of transcription after long exposures to light. The absence of the RCO-1/RCM-1 repressor complex leads to lower VVD levels that are available for the regulation of the activity of the WCC. The reduction in the amount of VVD results in increased WCC binding to the promoters of light-regulated genes in the dark and after long exposures to light, leading to the modification of photoadaptation that has been observed in rco-1 and rcm-1 mutants. Our results show that the photoadaptation phenotype of mutants in the RCO-1/RCM-1 repressor complex is, at least in part, an indirect consequence of the reduction of vvd transcription, and the resulting modification in the regulation of transcription by the WCC. [ABSTRACT FROM AUTHOR]
- Published
- 2014
- Full Text
- View/download PDF
24. Opposing Functions of Akt Isoforms in Lung Tumor Initiation and Progression.
- Author
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Linnerth-Petrik, Nicolle M., Santry, Lisa A., Petrik, James J., and Wootton, Sarah K.
- Subjects
- *
LUNG tumors , *PHOSPHATIDYLINOSITOL 3-kinases , *CELLULAR signal transduction , *PHOSPHOINOSITIDES , *LABORATORY mice , *CYTOLOGY , *MTOR protein - Abstract
Background: The phosphatidylinositol 3-kinase–regulated protein kinase, Akt, plays an important role in the initiation and progression of human cancer. Mammalian cells express three Akt isoforms (Akt1–3), which are encoded by distinct genes. Despite sharing a high degree of amino acid identity, phenotypes observed in knockout mice suggest that Akt isoforms are not functionally redundant. The relative contributions of the different Akt isoforms to oncogenesis, and the effect of their deficiencies on tumor development, are not well understood. Methods: Here we demonstrate that Akt isoforms have non-overlapping and sometimes opposing functions in tumor initiation and progression using a viral oncogene-induced mouse model of lung cancer and Akt isoform-specific knockout mice. Results: Akt1 ablation significantly delays initiation of lung tumor growth, whereas Akt2 deficiency dramatically accelerates tumorigenesis in this mouse model. Ablation of Akt3 had a small, not statistically significant, stimulatory effect on tumor induction and growth by the viral oncogene. Terminal deoxynucleotidyl transferase–mediated dUTP nick end labeling and Ki67 immunostaining of lung tissue sections revealed that the delayed tumor induction in Akt1−/− mice was due to the inhibitory effects of Akt1 ablation on cell growth and survival. Conversely, the accelerated growth rate of lung tumors in Akt2−/− and Akt3−/− mice was due to increased cell proliferation and reduced tumor cell apoptosis. Investigation of Akt signaling in tumors from Akt knockout mice revealed that the lack of Akt1 interrupted the propagation of signaling in tumors to the critical downstream targets, GSK-3α/β and mTOR. Conclusions: These results demonstrate that the degree of functional redundancy between Akt isoforms in the context of lung tumor initiation is minimal. Given that this mouse model exhibits considerable similarities to human lung cancer, these findings have important implications for the design and use of Akt inhibitors for the treatment of lung cancer. [ABSTRACT FROM AUTHOR]
- Published
- 2014
- Full Text
- View/download PDF
25. Alteration of Light-Dependent Gene Regulation by the Absence of the RCO-1/RCM-1 Repressor Complex in the Fungus Neurospora crassa.
- Author
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Ruger-Herreros, Carmen, Gil-Sánchez, María del Mar, Sancar, Gencer, Brunner, Michael, and Corrochano, Luis M.
- Subjects
TRANSCRIPTION factors ,GENETIC regulation ,PHOTORECEPTORS ,NEUROSPORA crassa ,CELLULAR signal transduction ,GENETIC repressors ,MESSENGER RNA - Abstract
The activation of transcription by light in the fungus Neurospora crassa requires the White Collar Complex (WCC), a photoreceptor and transcription factor complex. After light reception two WCCs interact and bind the promoters of light-regulated genes to activate transcription. This process is regulated by VVD, a small photoreceptor that disrupts the interaction between WCCs and leads to a reduction in transcription after long exposures to light. The N. crassa RCO-1/RCM-1 repressor complex is the homolog of the Tup1-Ssn6 repressor complex in yeast, and its absence modifies photoadaptation. We show that the absence of the RCO-1/RCM-1 repressor complex leads to several alterations in transcription that are gene-specific: an increase in the accumulation of mRNAs in the dark, a repression of transcription, and a derepression of transcription after long exposures to light. The absence of the RCO-1/RCM-1 repressor complex leads to lower VVD levels that are available for the regulation of the activity of the WCC. The reduction in the amount of VVD results in increased WCC binding to the promoters of light-regulated genes in the dark and after long exposures to light, leading to the modification of photoadaptation that has been observed in rco-1 and rcm-1 mutants. Our results show that the photoadaptation phenotype of mutants in the RCO-1/RCM-1 repressor complex is, at least in part, an indirect consequence of the reduction of vvd transcription, and the resulting modification in the regulation of transcription by the WCC. [ABSTRACT FROM AUTHOR]
- Published
- 2014
- Full Text
- View/download PDF
26. Light-Mediated Kinetic Control Reveals the Temporal Effect of the Raf/MEK/ERK Pathway in PC12 Cell Neurite Outgrowth.
- Author
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Zhang, Kai, Duan, Liting, Ong, Qunxiang, Lin, Ziliang, Varman, Pooja Mahendra, Sung, Kijung, and Cui, Bianxiao
- Subjects
- *
KINETIC control , *CELLULAR signal transduction , *CELL growth , *CHEMICAL biology , *EPIDERMAL growth factor , *CELL proliferation - Abstract
It has been proposed that differential activation kinetics allows cells to use a common set of signaling pathways to specify distinct cellular outcomes. For example, nerve growth factor (NGF) and epidermal growth factor (EGF) induce different activation kinetics of the Raf/MEK/ERK signaling pathway and result in differentiation and proliferation, respectively. However, a direct and quantitative linkage between the temporal profile of Raf/MEK/ERK activation and the cellular outputs has not been established due to a lack of means to precisely perturb its signaling kinetics. Here, we construct a light-gated protein-protein interaction system to regulate the activation pattern of the Raf/MEK/ERK signaling pathway. Light-induced activation of the Raf/MEK/ERK cascade leads to significant neurite outgrowth in rat PC12 pheochromocytoma cell lines in the absence of growth factors. Compared with NGF stimulation, light stimulation induces longer but fewer neurites. Intermittent on/off illumination reveals that cells achieve maximum neurite outgrowth if the off-time duration per cycle is shorter than 45 min. Overall, light-mediated kinetic control enables precise dissection of the temporal dimension within the intracellular signal transduction network. [ABSTRACT FROM AUTHOR]
- Published
- 2014
- Full Text
- View/download PDF
27. Bak Compensated for Bax in p53-null Cells to Release Cytochrome c for the Initiation of Mitochondrial Signaling during Withanolide D-Induced Apoptosis
- Subjects
Histology ,Physiology ,Mice, Nude ,Apoptosis ,Antineoplastic Agents ,Bioenergetics ,Research and Analysis Methods ,Biochemistry ,Mice ,Signal Initiation ,Spectrum Analysis Techniques ,Immune Physiology ,Cell Line, Tumor ,Medicine and Health Sciences ,Animals ,Humans ,Withanolides ,Energy-Producing Organelles ,bcl-2-Associated X Protein ,Cell Death ,Mechanisms of Signal Transduction ,Body Weight ,Biology and Life Sciences ,Cytochromes c ,Cell Biology ,Flow Cytometry ,Retraction ,Mitochondria ,Gene Expression Regulation, Neoplastic ,bcl-2 Homologous Antagonist-Killer Protein ,Physiological Parameters ,Cell Processes ,Spectrophotometry ,Cytophotometry ,Cellular Structures and Organelles ,Anatomy ,Tumor Suppressor Protein p53 ,K562 Cells ,Spleen ,Neoplasm Transplantation ,Signal Transduction - Abstract
The goal of cancer chemotherapy to induce multi-directional apoptosis as targeting a single pathway is unable to decrease all the downstream effect arises from crosstalk. Present study reports that Withanolide D (WithaD), a steroidal lactone isolated from Withania somnifera, induced cellular apoptosis in which mitochondria and p53 were intricately involved. In MOLT-3 and HCT116p53+/+ cells, WithaD induced crosstalk between intrinsic and extrinsic signaling through Bid, whereas in K562 and HCT116p53-/- cells, only intrinsic pathway was activated where Bid remain unaltered. WithaD showed pronounced activation of p53 in cancer cells. Moreover, lowered apoptogenic effect of HCT116p53-/- over HCT116p53+/+ established a strong correlation between WithaD-mediated apoptosis and p53. WithaD induced Bax and Bak upregulation in HCT116p53+/+, whereas increase only Bak expression in HCT116p53-/- cells, which was coordinated with augmented p53 expression. p53 inhibition substantially reduced Bax level and failed to inhibit Bak upregulation in HCT116p53+/+ cells confirming p53-dependent Bax and p53-independent Bak activation. Additionally, in HCT116p53+/+ cells, combined loss of Bax and Bak (HCT116Bax-Bak-) reduced WithaD-induced apoptosis and completely blocked cytochrome c release whereas single loss of Bax or Bak (HCT116Bax-Bak+/HCT116Bax+Bak-) was only marginally effective after WithaD treatment. In HCT116p53-/- cells, though Bax translocation to mitochondria was abrogated, Bak oligomerization helped the cells to release cytochrome c even before the disruption of mitochondrial membrane potential. WithaD also showed in vitro growth-inhibitory activity against an array of p53 wild type and null cancer cells and K562 xenograft in vivo. Taken together, WithaD elicited apoptosis in malignant cells through Bax/Bak dependent pathway in p53-wild type cells, whereas Bak compensated against loss of Bax in p53-null cells.
- Published
- 2020
28. Dissecting the dynamics of signaling events in the BMP, WNT, and NODAL cascade during self-organized fate patterning in human gastruloids
- Author
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Lizhong Liu, Aryeh Warmflash, Xiangyu Kong, Ryan Goh, and Sapna Chhabra
- Subjects
0301 basic medicine ,Embryology ,Cellular differentiation ,Human Embryonic Stem Cells ,Nodal signaling ,Bone Morphogenetic Protein 4 ,Mesoderm ,0302 clinical medicine ,Signal Initiation ,Cell Signaling ,Biology (General) ,WNT Signaling Cascade ,General Neuroscience ,Mechanisms of Signal Transduction ,Wnt signaling pathway ,Cell Differentiation ,Signaling Cascades ,Cell biology ,Organoids ,medicine.anatomical_structure ,embryonic structures ,General Agricultural and Biological Sciences ,Research Article ,Signal Transduction ,BMP signaling ,Signal Inhibition ,animal structures ,Nodal Protein ,QH301-705.5 ,SMAD signaling ,Biology ,Models, Biological ,General Biochemistry, Genetics and Molecular Biology ,Cell Line ,03 medical and health sciences ,Paracrine signalling ,medicine ,Humans ,Benzothiazoles ,Body Patterning ,Models, Statistical ,General Immunology and Microbiology ,Embryos ,Gastrulation ,Biology and Life Sciences ,Cell Biology ,Gastrula ,Wnt Proteins ,030104 developmental biology ,Gene Expression Regulation ,Epiblast ,NODAL ,030217 neurology & neurosurgery ,Developmental Biology - Abstract
During gastrulation, the pluripotent epiblast self-organizes into the 3 germ layers—endoderm, mesoderm and ectoderm, which eventually form the entire embryo. Decades of research in the mouse embryo have revealed that a signaling cascade involving the Bone Morphogenic Protein (BMP), WNT, and NODAL pathways is necessary for gastrulation. In vivo, WNT and NODAL ligands are expressed near the site of gastrulation in the posterior of the embryo, and knockout of these ligands leads to a failure to gastrulate. These data have led to the prevailing view that a signaling gradient in WNT and NODAL underlies patterning during gastrulation; however, the activities of these pathways in space and time have never been directly observed. In this study, we quantify BMP, WNT, and NODAL signaling dynamics in an in vitro model of human gastrulation. Our data suggest that BMP signaling initiates waves of WNT and NODAL signaling activity that move toward the colony center at a constant rate. Using a simple mathematical model, we show that this wave-like behavior is inconsistent with a reaction-diffusion–based Turing system, indicating that there is no stable signaling gradient of WNT/NODAL. Instead, the final signaling state is homogeneous, and spatial differences arise only from boundary effects. We further show that the durations of WNT and NODAL signaling control mesoderm differentiation, while the duration of BMP signaling controls differentiation of CDX2-positive extra-embryonic cells. The identity of these extra-embryonic cells has been controversial, and we use RNA sequencing (RNA-seq) to obtain their transcriptomes and show that they closely resemble human trophoblast cells in vivo. The domain of BMP signaling is identical to the domain of differentiation of these trophoblast-like cells; however, neither WNT nor NODAL forms a spatial pattern that maps directly to the mesodermal region, suggesting that mesoderm differentiation is controlled dynamically by the combinatorial effect of multiple signals. We synthesize our data into a mathematical model that accurately recapitulates signaling dynamics and predicts cell fate patterning upon chemical and physical perturbations. Taken together, our study shows that the dynamics of signaling events in the BMP, WNT, and NODAL cascade in the absence of a stable signaling gradient control fate patterning of human gastruloids., A self-organizing in vitro model of cell fate decisions during human gastrulation reveals that patterning is controlled by combinatorial dynamics of multiple signaling pathways, rather than by stable gradients.
- Published
- 2019
29. Stimulation of α1a Adrenergic Receptors Induces Cellular Proliferation or Antiproliferative Hypertrophy Dependent Solely on Agonist Concentration.
- Author
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Lei, Beilei, Schwinn, Debra A., and Morris, Daniel P.
- Subjects
- *
ADRENERGIC receptors , *CELL proliferation , *HYPERTROPHY , *LABORATORY rats , *PHENYLEPHRINE , *EPIDERMAL growth factor receptors , *EUKARYOTIC cells - Abstract
Stimulation of α1aAdrenergic Receptors (ARs) is known to have anti-proliferative and hypertrophic effects; however, some studies also suggests this receptor can increase cell proliferation. Surprisingly, we find the α1aAR expressed in rat-1 fibroblasts can produce either phenotype, depending exclusively on agonist concentration. Stimulation of the α1aAR by high dose phenylephrine (>10−7 M) induces an antiproliferative, hypertrophic response accompanied by robust and extended p38 activation. Inhibition of p38 with SB203580 prevented the antiproliferative response, while inhibition of Erk or Jnk had no effect. In stark contrast, stimulation of the α1aAR with low dose phenylephrine (∼10−8 M) induced an Erk-dependent increase in cellular proliferation. Agonist-induced Erk phosphorylation was preceded by rapid FGFR and EGFR transactivation; however, only EGFR inhibition blocked Erk activation and proliferation. The general matrix metalloprotease inhibitor, GM6001, blocked agonist induced Erk activation within seconds, strongly suggesting EGFR activation involved extracellular triple membrane pass signaling. Erk activation required little Ca2+ release and was blocked by PLCβ or PKC inhibition but not by intracellular Ca2+ chelation, suggesting Ca2+ independent activation of novel PKC isoforms. In contrast, Ca2+ release was essential for PI3K/Akt activation, which was acutely maximal at non-proliferative doses of agonist. Remarkably, our data suggests EGFR transactivation leading to Erk induced proliferation has the lowest activation threshold of any α1aAR response. The ability of α1aARs to induce proliferation are discussed in light of evidence suggesting antagonistic growth responses reflect native α1aAR function. [ABSTRACT FROM AUTHOR]
- Published
- 2013
- Full Text
- View/download PDF
30. EphA2-Induced Angiogenesis in Ewing Sarcoma Cells Works through bFGF Production and Is Dependent on Caveolin-1.
- Author
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Sáinz-Jaspeado, Miguel, Huertas-Martinez, Juan, Lagares-Tena, Laura, Martin Liberal, Juan, Mateo-Lozano, Silvia, de Alava, Enrique, de Torres, Carmen, Mora, Jaume, Muro, Xavier Garcia del, and Tirado, Oscar M.
- Subjects
- *
EPHRIN receptors , *PROTEIN-tyrosine kinases , *NEOVASCULARIZATION , *SARCOMA , *CANCER cells , *FIBROBLAST growth factors , *CAVEOLINS , *CELLULAR signal transduction , *TUMORS in children - Abstract
Angiogenesis is the result of the combined activity of the tumor microenvironment and signaling molecules. The angiogenic switch is represented as an imbalance between pro- and anti-angiogenic factors and is a rate-limiting step in the development of tumors. Eph receptor tyrosine kinases and their membrane-anchored ligands, known as ephrins, constitute the largest receptor tyrosine kinase (RTK) subfamily and are considered a major family of pro-angiogenic RTKs. Ewing sarcoma (EWS) is a highly aggressive bone and soft tissue tumor affecting children and young adults. As other solid tumors, EWS are reliant on a functional vascular network for the delivery of nutrients and oxygen and for the removal of waste. Based on the biological roles of EphA2 in promoting angiogenesis, we explored the functional role of this receptor and its relationship with caveolin-1 (CAV1) in EWS angiogenesis. We demonstrated that lack of CAV1 results in a significant reduction in micro vascular density (MVD) on 3 different in vivo models. In vitro, this phenomenon correlated with inactivation of EphA2 receptor, lack of AKT response and downregulation of bFGF. We also demonstrated that secreted bFGF from EWS cells acted as chemoattractant for endothelial cells. Furthermore, interaction between EphA2 and CAV1 was necessary for the right localization and signaling of the receptor to produce bFGF through AKT and promote migration of endothelial cells. Finally, introduction of a dominant-negative form of EphA2 into EWS cells mostly reproduced the effects occurred by CAV1 silencing, strongly suggesting that the axis EphA2-CAV1 participates in the promotion of endothelial cell migration toward the tumors favoring EWS angiogenesis. [ABSTRACT FROM AUTHOR]
- Published
- 2013
- Full Text
- View/download PDF
31. NCI-H295R, a Human Adrenal Cortex-Derived Cell Line, Expresses Purinergic Receptors Linked to Ca2+-Mobilization/Influx and Cortisol Secretion.
- Author
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Nishi, Haruhisa, Arai, Hirokazu, and Momiyama, Toshihiko
- Subjects
- *
ADRENAL cortex , *CELL lines , *PURINERGIC receptors , *HYDROCORTISONE , *SECRETION , *MESSENGER RNA - Abstract
Purinergic receptor expression and involvement in steroidogenesis were examined in NCI-H295R (H295R), a human adrenal cortex cell line which expresses all the key enzymes necessary for steroidogenesis. mRNA/protein for multiple P1 (A2A and A2B), P2X (P2X5 and P2X7), and P2Y (P2Y1, P2Y2, P2Y6, P2Y12, P2Y13, and P2Y14) purinergic receptors were detected in H295R. 2MeS-ATP (10–1000 µM), a P2Y1 agonist, induced glucocorticoid (GC) secretion in a dose-dependent manner, while other extracellular purine/pyrimidine agonists (1–1000 µM) had no distinct effect on GC secretion. Extracellular purines, even non-steroidogenic ones, induced Ca2+-mobilization in the cells, independently of the extracellular Ca2+ concentration. Increases in intracellular Ca2+ concentration induced by extracellular purine agonists were transient, except when induced by ATP or 2MeS-ATP. Angiotensin II (AngII: 100 nM) and dibutyryl-cyclic AMP (db-cAMP: 500 µM) induced both GC secretion and Ca2+-mobilization in the presence of extracellular Ca2+ (1.2 mM). GC secretion by AngII was reduced by nifedipine (10–100 µM); whereas the Ca2+ channel blocker did not inhibit GC secretion by 2MeS-ATP. Thapsigargin followed by extracellular Ca2+ exposure induced Ca2+-influx in H295R, and the cells expressed mRNA/protein of the component molecules for store-operated calcium entry (SOCE): transient receptor C (TRPC) channels, calcium release-activated calcium channel protein 1 (Orai-1), and the stromal interaction molecule 1 (STIM1). In P2Y1-knockdown, 2MeS-ATP-induced GC secretion was significantly inhibited. These results suggest that H295R expresses a functional P2Y1 purinergic receptor for intracellular Ca2+-mobilization, and that P2Y1 is linked to SOCE-activation, leading to Ca2+-influx which might be necessary for glucocorticoid secretion. [ABSTRACT FROM AUTHOR]
- Published
- 2013
- Full Text
- View/download PDF
32. Modulation of the Akt Pathway Reveals a Novel Link with PERK/eIF2α, which Is Relevant during Hypoxia.
- Author
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Blaustein, Matías, Pérez-Munizaga, Daniela, Sánchez, Manuel Alejandro, Urrutia, Carolina, Grande, Alicia, Risso, Guillermo, Srebrow, Anabella, Alfaro, Jennifer, and Colman-Lerner, Alejandro
- Subjects
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PROTEIN kinase B , *BIOCHEMICAL research , *CELLULAR signal transduction , *ONCOGENES , *GROWTH factors , *CELL death - Abstract
The unfolded protein response (UPR) and the Akt signaling pathway share several regulatory functions and have the capacity to determine cell outcome under specific conditions. However, both pathways have largely been studied independently. Here, we asked whether the Akt pathway regulates the UPR. To this end, we used a series of chemical compounds that modulate PI3K/Akt pathway and monitored the activity of the three UPR branches: PERK, IRE1 and ATF6. The antiproliferative and antiviral drug Akt-IV strongly and persistently activated all three branches of the UPR. We present evidence that activation of PERK/eIF2α requires Akt and that PERK is a direct Akt target. Chemical activation of this novel Akt/PERK pathway by Akt-IV leads to cell death, which was largely dependent on the presence of PERK and IRE1. Finally, we show that hypoxia-induced activation of eIF2α requires Akt, providing a physiologically relevant condition for the interaction between Akt and the PERK branch of the UPR. These data suggest the UPR and the Akt pathway signal to one another as a means of controlling cell fate. [ABSTRACT FROM AUTHOR]
- Published
- 2013
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33. S100A8/A9 (Calprotectin) Negatively Regulates G2/M Cell Cycle Progression and Growth of Squamous Cell Carcinoma.
- Author
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Khammanivong, Ali, Wang, Chengxing, Sorenson, Brent S., Ross, Karen F., and Herzberg, Mark C.
- Subjects
- *
SQUAMOUS cell carcinoma , *CANCER invasiveness , *BIOCHEMICAL models , *CELL cycle regulation , *TUMOR growth , *HEAD & neck cancer , *CALCIUM-binding proteins - Abstract
Malignant transformation results in abnormal cell cycle regulation and uncontrolled growth in head and neck squamous cell carcinoma (HNSCC) and other cancers. S100A8/A9 (calprotectin) is a calcium-binding heterodimeric protein complex implicated in cell cycle regulation, but the specific mechanism and role in cell cycle control and carcinoma growth are not well understood. In HNSCC, S100A8/A9 is downregulated at both mRNA and protein levels. We now report that downregulation of S100A8/A9 correlates strongly with a loss of cell cycle control and increased growth of carcinoma cells. To show its role in carcinogenesis in an in vitro model, S100A8/A9 was stably expressed in an S100A8/A9-negative human carcinoma cell line (KB cells, HeLa-like). S100A8/A9 expression increases PP2A phosphatase activity and p-Chk1 (Ser345) phosphorylation, which appears to signal inhibitory phosphorylation of mitotic p-Cdc25C (Ser216) and p-Cdc2 (Thr14/Tyr15) to inactivate the G2/M Cdc2/cyclin B1 complex. Cyclin B1 expression then downregulates and the cell cycle arrests at the G2/M checkpoint, reducing cell division. As expected, S100A8/A9-expressing cells show both decreased anchorage-dependent and -independent growth and mitotic progression. Using shRNA, silencing of S100A8/A9 expression in the TR146 human HNSCC cell line increases growth and survival and reduces Cdc2 inhibitory phosphorylation at Thr14/Tyr15. The level of S100A8/A9 endogenous expression correlates strongly with the reduced p-Cdc2 (Thr14/Tyr14) level in HNSCC cell lines, SCC-58, OSCC-3 and UMSCC-17B. S100A8/A9-mediated control of the G2/M cell cycle checkpoint is, therefore, a likely suppressive mechanism in human squamous cell carcinomas and may suggest new therapeutic approaches. [ABSTRACT FROM AUTHOR]
- Published
- 2013
- Full Text
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34. Magnetic Nanoparticles as Mediators of Ligand-Free Activation of EGFR Signaling.
- Author
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Bharde, Atul A., Palankar, Raghavendra, Fritsch, Cornelia, Klaver, Arjen, Kanger, Johannes S., Jovin, Thomas M., and Arndt-Jovin, Donna J.
- Subjects
- *
MAGNETIC nanoparticles , *EPIDERMAL growth factor receptors , *CELLULAR signal transduction , *CANCER treatment , *LIGANDS (Biochemistry) , *MEDICAL research , *IRON oxides - Abstract
Background: Magnetic nanoparticles (NPs) are of particular interest in biomedical research, and have been exploited for molecular separation, gene/drug delivery, magnetic resonance imaging, and hyperthermic cancer therapy. In the case of cultured cells, magnetic manipulation of NPs provides the means for studying processes induced by mechanotransduction or by local clustering of targeted macromolecules, e.g. cell surface receptors. The latter are normally activated by binding of their natural ligands mediating key signaling pathways such as those associated with the epidermal growth factor (EGFR). However, it has been reported that EGFR may be dimerized and activated even in the absence of ligands. The present study assessed whether receptor clustering induced by physical means alone suffices for activating EGFR in quiescent cells. Methodology/Principal Findings: The EGFR on A431 cells was specifically targeted by superparamagnetic iron oxide NPs (SPIONs) carrying either a ligand-blocking monoclonal anti-EGFR antibody or a streptavidin molecule for targeting a chimeric EGFR incorporating a biotinylated amino-terminal acyl carrier peptide moiety. Application of a magnetic field led to SPION magnetization and clustering, resulting in activation of the EGFR, a process manifested by auto and transphosphorylation and downstream signaling. The magnetically-induced early signaling events were similar to those inherent to the ligand dependent EGFR pathways. Magnetization studies indicated that the NPs exerted magnetic dipolar forces in the sub-piconewton range with clustering dependent on Brownian motion of the receptor-SPION complex and magnetic field strength. Conclusions/Significance: We demonstrate that EGFR on the cell surface that have their ligand binding-pocket blocked by an antibody are still capable of transphosphorylation and initiation of signaling cascades if they are clustered by SPIONs either attached locally or targeted to another site of the receptor ectodomain. The results suggest that activation of growth factor receptors may be triggered by ligand-independent molecular crowding resulting from overexpression and/or sequestration in membrane microdomains. [ABSTRACT FROM AUTHOR]
- Published
- 2013
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35. HSP-72 Accelerated Expression in Mononuclear Cells Induced In Vivo by Acetyl Salicylic Acid Can Be Reproduced In Vitro when Combined with H2O2.
- Author
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Sandoval-Montiel, Alvaro A., Zentella-de-Piña, Martha, Ventura-Gallegos, José L., Frías-González, Susana, López-Macay, Ambar, and Zentella-Dehesa, Alejandro
- Subjects
- *
HEAT shock proteins , *GENE expression , *MONONUCLEOSIS , *SALICYLIC acid , *MESSENGER RNA , *LABORATORY rats , *PHYSIOLOGICAL effects of hydrogen peroxide - Abstract
Background: Among NSAIDs acetyl salicylic acid remains as a valuable tool because of the variety of benefic prophylactic and therapeutic effects. Nevertheless, the molecular bases for these responses have not been complete understood. We explored the effect of acetyl salicylic acid on the heat shock response. Results: Peripheral blood mononuclear cells from rats challenged with acetyl salicylic acid presented a faster kinetics of expression of HSP-72 messenger RNA and protein in response to in vitro heat shock. This effect reaches its maximum 2 h after treatment and disappeared after 5 h. On isolated peripheral blood mononuclear cells from untreated rats, incubation with acetyl salicylic acid was ineffective to produce priming, but this effect was mimicked when the cells were incubated with the combination of H2O2+ ASA. Conclusions: Administration of acetyl salicylic acid to rats alters HSP-72 expression mechanism in a way that it becomes more efficient in response to in vitro heat shock. The fact that in vitro acetyl salicylic acid alone did not induce this priming effect implies that in vivo other signals are required. Priming could be reproduces in vitro with the combination of acetyl salicylic acid+H2O2. [ABSTRACT FROM AUTHOR]
- Published
- 2013
- Full Text
- View/download PDF
36. HSP-72 Accelerated Expression in Mononuclear Cells Induced In Vivo by Acetyl Salicylic Acid Can Be Reproduced In Vitro when Combined with H2O2.
- Author
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Sandoval-Montiel, Alvaro A., Zentella-de-Piña, Martha, Ventura-Gallegos, José L., Frías-González, Susana, López-Macay, Ambar, and Zentella-Dehesa, Alejandro
- Subjects
HEAT shock proteins ,GENE expression ,MONONUCLEOSIS ,SALICYLIC acid ,MESSENGER RNA ,LABORATORY rats ,PHYSIOLOGICAL effects of hydrogen peroxide - Abstract
Background: Among NSAIDs acetyl salicylic acid remains as a valuable tool because of the variety of benefic prophylactic and therapeutic effects. Nevertheless, the molecular bases for these responses have not been complete understood. We explored the effect of acetyl salicylic acid on the heat shock response. Results: Peripheral blood mononuclear cells from rats challenged with acetyl salicylic acid presented a faster kinetics of expression of HSP-72 messenger RNA and protein in response to in vitro heat shock. This effect reaches its maximum 2 h after treatment and disappeared after 5 h. On isolated peripheral blood mononuclear cells from untreated rats, incubation with acetyl salicylic acid was ineffective to produce priming, but this effect was mimicked when the cells were incubated with the combination of H
2 O2 + ASA. Conclusions: Administration of acetyl salicylic acid to rats alters HSP-72 expression mechanism in a way that it becomes more efficient in response to in vitro heat shock. The fact that in vitro acetyl salicylic acid alone did not induce this priming effect implies that in vivo other signals are required. Priming could be reproduces in vitro with the combination of acetyl salicylic acid+H2 O2 . [ABSTRACT FROM AUTHOR]- Published
- 2013
- Full Text
- View/download PDF
37. The Second Intracellular Loop of the Human Cannabinoid CB2 Receptor Governs G Protein Coupling in Coordination with the Carboxyl Terminal Domain
- Author
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Zheng, Congxia, Chen, Linjie, Chen, Xiaopan, He, Xiaobai, Yang, Jingwen, Shi, Ying, and Zhou, Naiming
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- *
CANNABINOID receptors , *INTRACELLULAR pathogens , *G protein coupled receptors , *CARBOXYL group , *MEMBRANE proteins , *CELLULAR signal transduction , *ADENYLATE cyclase - Abstract
The major effects of cannabinoids and endocannabinoids are mediated via two G protein-coupled receptors, CB1 and CB2, elucidation of the mechanism and structural determinants of the CB2 receptor coupling with G proteins will have a significant impact on drug discovery. In the present study, we systematically investigated the role of the intracellular loops in the interaction of the CB2 receptor with G proteins using chimeric receptors alongside the characterization of cAMP accumulation and ERK1/2 phosphorylation. We provided evidence that ICL2 was significantly involved in G protein coupling in coordination with the C-terminal end. Moreover, a single alanine substitution of the Pro-139 in the CB2 receptor that corresponds to Leu-222 in the CB1 receptor resulted in a moderate impairment in the inhibition of cAMP accumulation, whereas mutants P139F, P139M and P139L were able to couple to the Gs protein in a CRE-driven luciferase assay. With the ERK activation experiments, we further found that P139L has the ability to activate ERK through both Gi- and Gs-mediated pathways. Our findings defined an essential role of the second intracellular loop of the CB2 receptor in coordination with the C-terminal tail in G protein coupling and receptor activation. [ABSTRACT FROM AUTHOR]
- Published
- 2013
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38. LKB1 Mediates the Development of Conventional and Innate T Cells via AMP-Dependent Kinase Autonomous Pathways.
- Author
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Zarrouk, Marouan, Rolf, Julia, and Cantrell, Doreen Ann
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T cell receptors , *NATURAL immunity , *MITOGEN-activated protein kinases , *SERINE/THREONINE kinases , *THYMOCYTES , *ANTIGEN receptors , *CELL differentiation , *CELLULAR signal transduction - Abstract
The present study has examined the role of the serine/threonine kinase LKB1 in the survival and differentiation of CD4/8 double positive thymocytes. LKB1-null DPs can respond to signals from the mature α/β T-cell-antigen receptor and initiate positive selection. However, in the absence of LKB1, thymocytes fail to mature to conventional single positive cells causing severe lymphopenia in the peripheral lymphoid tissues. LKB1 thus appears to be dispensable for positive selection but important for the maturation of positively selected thymocytes. LKB1 also strikingly prevented the development of invariant Vα14 NKT cells and innate TCR αβ gut lymphocytes. Previous studies with gain of function mutants have suggested that the role of LKB1 in T cell development is mediated by its substrate the AMP-activated protein kinase (AMPK). The present study now analyses the impact of AMPK deletion in DP thymocytes and shows that the role of LKB1 during the development of both conventional and innate T cells is mediated by AMPK-independent pathways. [ABSTRACT FROM AUTHOR]
- Published
- 2013
- Full Text
- View/download PDF
39. Insulin-Like Growth Factor Binding Proteins Increase Intracellular Calcium Levels in Two Different Cell Lines.
- Author
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Seurin, Danielle, Lombet, Alain, Babajko, Sylvie, Godeau, François, and Ricort, Jean-Marc
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- *
INSULIN-like growth factor-binding proteins , *INTRACELLULAR calcium , *CELL lines , *CELL differentiation , *CELL growth , *APOPTOSIS , *CELLULAR signal transduction , *FLOW cytometry - Abstract
Background: Insulin-like growth factor binding proteins (IGFBPs) are six related secreted proteins that share IGF-dependent and -independent functions. If the former functions begin to be well described, the latter are somewhat more difficult to investigate and to characterize. At the cellular level, IGFBPs were shown to modulate numerous processes including cell growth, differentiation and apoptosis. However, the molecular mechanisms implicated remain largely unknown. We previously demonstrated that IGFBP-3, but not IGFBP-1 or IGFBP-5, increase intracellular calcium concentration in MCF-7 cells (Ricort J-M et al. (2002) FEBS lett 527: 293–297). Methodology/Principal Findings: We perform a global analysis in which we studied, by two different approaches, the binding of each IGFBP isoform (i.e., IGFBP-1 to -6) to the surface of two different cellular models, MCF-7 breast adenocarcinoma cells and C2 myoblast proliferative cells, as well as the IGFBP-induced increase of intracellular calcium concentration. Using both confocal fluorescence microscopy and flow cytometry analysis, we showed that all IGFBPs bind to MCF-7 cell surface. By contrast, only four IGFBPs can bind to C2 cell surface since neither IGFBP-2 nor IGFBP-4 were detected. Among the six IGFBPs tested, only IGFBP-1 did not increased intracellular calcium concentration whatever the cellular model studied. By contrast, IGFBP-2, -3, -4 and -6, in MCF-7 cells, and IGFBP-3, -5 and -6, in C2 proliferative cells, induce a rapid and transient increase in intracellular free calcium concentration. Moreover, IGFBP-2 and -3 (in MCF-7 cells) and IGFBP-5 (in C2 cells) increase intracellular free calcium concentration by a pertussis toxin sensitive signaling pathway. Conclusions: Our results demonstrate that IGFBPs are able to bind to cell surface and increase intracellular calcium concentration. By characterizing the IGFBPs-induced cell responses and intracellular couplings, we highlight the cellular specificity and complexity of the IGF-independent actions of these IGF binding proteins. [ABSTRACT FROM AUTHOR]
- Published
- 2013
- Full Text
- View/download PDF
40. P38/NF-κB/Snail Pathway Is Involved in Caffeic Acid-Induced Inhibition of Cancer Stem Cells-Like Properties and Migratory Capacity in Malignant Human Keratinocyte.
- Author
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Yang, Ye, Li, Yuan, Wang, Kebo, Wang, Yu, Yin, Wenqin, and Li, Lei
- Subjects
- *
CANCER treatment , *SKIN cancer , *NF-kappa B , *EPITHELIAL cells , *METASTASIS , *CANCER stem cells , *KERATINOCYTES , *ANTINEOPLASTIC agents , *CELLULAR signal transduction , *CANCER cells - Abstract
Background: Skin cancer is the most common cancer throughout the world. The epithelial-mesenchymal transition (EMT) and the acquisition of cancer stem cells (CSCs)-like properties emerge as critical steps in the metastasis of human skin cancers. Caffeic acid (CaA) exerts anticarcinogenic effects. However, the effects of CaA on the migratory capability and on the CSCs-like properties of skin cancer cells, and the molecular mechanisms underlying it are not fully understood. Methods: Malignant HaCaT cells were treated by CaA. Transwell assay was performed to determine that CaA attenuated the migratory capability; Spheroid formation assay was performed to confirm that CaA decreased the CSCs-like phenotype; Treated malignant HaCaT cells were molecularly characterized by RT-PCR, Western blots, Southwestern blot, and immunoprecipitation. Results: In CaA-treated malignant human keratinocyte (malignant HaCaT cells), inhibition of the migratory capability and CSCs-like phenotype were observed. CaA up-regulated the phosphorylation of p38, and down-regulated the activation of nuclear factor κB (NF-κB)/snail signal pathway. Indeed, p38 decreased the DNA-binding activity of NF-κB to the promoter of snail gene, which resulted in the transcriptional inactivation of snail. Blockage of p38 attenuated the CaA-induced inhibition of migratory capability and CSCs-like phenotype in malignant HaCaT cells. Conclusions: CaA attenuates the migratory capability and CSCs-like Properties of malignant human keratinocyte, in which, p38-mediated down-regulation of NF-κB/snail signal pathway is involved. [ABSTRACT FROM AUTHOR]
- Published
- 2013
- Full Text
- View/download PDF
41. Akt Regulates TNFα Synthesis Downstream of RIP1 Kinase Activation during Necroptosis.
- Author
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McNamara, Colleen R., Ahuja, Ruchita, Osafo-Addo, Awo D., Barrows, Douglas, Kettenbach, Arminja, Skidan, Igor, Teng, Xin, Cuny, Gregory D., Gerber, Scott, and Degterev, Alexei
- Subjects
- *
PROTEIN kinase B , *TUMOR necrosis factors , *RECEPTOR-interacting proteins , *CELL death , *NECROSIS , *INFLAMMATORY bowel diseases , *PHOSPHORYLATION , *CELLULAR signal transduction , *RAPAMYCIN , *MACROPHAGES - Abstract
Necroptosis is a regulated form of necrotic cell death that has been implicated in the pathogenesis of various diseases including intestinal inflammation and systemic inflammatory response syndrome (SIRS). In this work, we investigated the signaling mechanisms controlled by the necroptosis mediator receptor interacting protein-1 (RIP1) kinase. We show that Akt kinase activity is critical for necroptosis in L929 cells and plays a key role in TNFα production. During necroptosis, Akt is activated in a RIP1 dependent fashion through its phosphorylation on Thr308. In L929 cells, this activation requires independent signaling inputs from both growth factors and RIP1. Akt controls necroptosis through downstream targeting of mammalian Target of Rapamycin complex 1 (mTORC1). Akt activity, mediated in part through mTORC1, links RIP1 to JNK activation and autocrine production of TNFα. In other cell types, such as mouse lung fibroblasts and macrophages, Akt exhibited control over necroptosis-associated TNFα production without contributing to cell death. Overall, our results provide new insights into the mechanism of necroptosis and the role of Akt kinase in both cell death and inflammatory regulation. [ABSTRACT FROM AUTHOR]
- Published
- 2013
- Full Text
- View/download PDF
42. Role of the Phosphatase PTEN in Early Vascular Remodeling.
- Author
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Sedding, Daniel G., Widmer-Teske, Rebecca, Mueller, Andreas, Stieger, Philipp, Daniel, Jan-Marcus, Gündüz, Dursun, Pullamsetti, Soni, Nef, Holger, Moellmann, Helge, Troidl, Christian, Hamm, Christian, and Braun-Dullaeus, Rüdiger
- Subjects
- *
PHOSPHATASES , *PTEN protein , *TISSUE remodeling , *PHOSPHATIDYLINOSITOLS , *BLOOD vessels , *WOUNDS & injuries , *PROTEIN kinase B , *CELLULAR signal transduction , *ANGIOPLASTY , *IN vitro studies - Abstract
Background: The phosphatase PTEN represents an important physiological inhibitor of phosphatidylinositol-3 kinase (PI3-K)/protein kinase B (Akt) signalling, however, the functional role of PTEN in the initial phase of angioplasty-induced vascular injury remains elusive. In the present study we sought to determine PTEN's effect on vascular smooth muscle cell (VSMC) apoptosis following acute injury in vivo and in vitro. Methods and Results: Immunohistochemistry indicated a faint basal expression and equal distribution of PTEN in uninjured rat carotid arteries. 12 h following balloon-injury, PTEN expression was strongly increased in apoptotic (TUNEL+) VSMC. In vitro, stimulation with serum or different growth factors or subjecting VSMC to cyclic stretch had no effect on PTEN expression, whereas stimulation with H2O2 robustly increased PTEN expression in a time- and dose-dependent manner. To evaluate the functional role of PTEN expression, human VSMC were transduced with WT-PTEN. Overexpression of PTEN increased the number of apoptotic VSMC (19.8%±4.4 vs. 5.6%±2.3; P<0.001) as determined by TUNEL assay. In contrast, siRNA-mediated knock-down of PTEN attenuated the basal as well as H2O2-induced apoptosis of VSMC. Mechanistically, overexpression of PTEN prevented serum-induced Akt-phosphorylation, whereas siRNA-mediated knock down of PTEN augmented Akt-activation. Moreover, co-transfection of PTEN and a constitutive active Akt mutant prevented PTEN-dependent augmentation of VSMC apoptosis, indicating, that PTEN regulates VSMC apoptosis by inhibition of Akt phosphorylation/activation. Conclusion: By interfering with the PI3-K/Akt-dependent survival signalling, the oxidative stress-induced up regulation of PTEN in VSMC of injured arteries augments the sensitivity of VSMC to apoptotic stimuli in the early phase following vascular injury, augmenting the initial injury and cell loss of the injured vessel wall. Thus, these data add to our understanding of PTEN's role during vascular remodelling. [ABSTRACT FROM AUTHOR]
- Published
- 2013
- Full Text
- View/download PDF
43. Killing two birds with one stone: How the respiratory syncytial virus polymerase initiates transcription and replication
- Author
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Sarah L. Noton, Chadene Z. Tremaglio, and Rachel Fearns
- Subjects
RNA viruses ,Transcription, Genetic ,Human metabolism ,Pathology and Laboratory Medicine ,Virus Replication ,Biochemistry ,Polymerases ,Pearls ,Viral Packaging ,Signal Initiation ,Cell Signaling ,Transcription (biology) ,Medicine and Health Sciences ,Respiratory system ,Biology (General) ,Polymerase ,0303 health sciences ,biology ,Messenger RNA ,030302 biochemistry & molecular biology ,Mechanisms of Signal Transduction ,Respiratory Syncytial Viruses ,Nucleic acids ,Medical Microbiology ,Viral Pathogens ,Viruses ,Pathogens ,Genomic Signal Processing ,Signal Transduction ,DNA Replication ,QH301-705.5 ,Nucleic acid synthesis ,Immunology ,Respiratory Syncytial Virus Infections ,Microbiology ,Virus ,03 medical and health sciences ,Virology ,DNA-binding proteins ,Genetics ,Humans ,Chemical synthesis ,RNA synthesis ,Molecular Biology ,Microbial Pathogens ,030304 developmental biology ,Biology and life sciences ,Organisms ,Proteins ,Human physiology ,Cell Biology ,RC581-607 ,RNA-Dependent RNA Polymerase ,Viral Replication ,Research and analysis methods ,Biosynthetic techniques ,Viral replication ,Respiratory Syncytial Virus, Human ,Paramyxoviruses ,biology.protein ,RNA ,Parasitology ,Respiratory Syncytial Virus ,Immunologic diseases. Allergy - Published
- 2019
44. MicroRNAs as possible indicators of drug sensitivity in breast cancer cell lines
- Author
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John A. Foekens, Agnes Jager, John W.M. Martens, Anouk A. J. Heine, Marijn T.M. van Jaarsveld, Erik A.C. Wiemer, Wendy J. C. Prager-van der Smissen, Antonius W. M. Boersma, Bahar Ozturk, Marcel Smid, Katharina Uhr, and Medical Oncology
- Subjects
0301 basic medicine ,Drug resistance ,Biochemistry ,chemistry.chemical_compound ,0302 clinical medicine ,Signal Initiation ,Cell Signaling ,Breast Tumors ,Medicine and Health Sciences ,Medicine ,RNA, Neoplasm ,Cell Cycle and Cell Division ,media_common ,Multidisciplinary ,Mechanisms of Signal Transduction ,3. Good health ,G2 Phase Cell Cycle Checkpoints ,Nucleic acids ,Paclitaxel ,Docetaxel ,Oncology ,Cell Processes ,030220 oncology & carcinogenesis ,embryonic structures ,Female ,Network Analysis ,medicine.drug ,Research Article ,Signal Transduction ,Drug ,Computer and Information Sciences ,Signal Inhibition ,Veliparib ,Science ,media_common.quotation_subject ,Antineoplastic Agents ,Breast Neoplasms ,03 medical and health sciences ,Breast cancer ,SDG 3 - Good Health and Well-being ,Panobinostat ,Cell Line, Tumor ,Breast Cancer ,Biomarkers, Tumor ,Genetics ,Humans ,Tivantinib ,Non-coding RNA ,Pharmacology ,Natural antisense transcripts ,Drug Screening ,Biology and life sciences ,business.industry ,Cancers and Neoplasms ,Cell Biology ,medicine.disease ,Gene regulation ,Signaling Networks ,body regions ,MicroRNAs ,030104 developmental biology ,chemistry ,Drug Resistance, Neoplasm ,Cancer research ,M Phase Cell Cycle Checkpoints ,RNA ,Gene expression ,business ,Biomarkers - Abstract
MicroRNAs (miRNAs) regulate gene expression post-transcriptionally. In this way they might influence whether a cell is sensitive or resistant to a certain drug. So far, only a limited number of relatively small scale studies comprising few cell lines and/or drugs have been performed. To obtain a broader view on miRNAs and their association with drug response, we investigated the expression levels of 411 miRNAs in relation to drug sensitivity in 36 breast cancer cell lines. For this purpose IC50 values of a drug screen involving 34 drugs were associated with miRNA expression data of the same breast cancer cell lines. Since molecular subtype of the breast cancer cell lines is considered a confounding factor in drug association studies, multivariate analysis taking subtype into account was performed on significant miRNA-drug associations which retained 13 associations. These associations consisted of 11 different miRNAs and eight different drugs (among which Paclitaxel, Docetaxel and Veliparib). The taxanes, Paclitaxel and Docetaxel, were the only drugs having miRNAs in common: hsa-miR-187-5p and hsa-miR-106a-3p indicative of drug resistance while Paclitaxel sensitivity alone associated with hsa-miR-556-5p. Tivantinib was associated with hsa-let-7d-5p and hsa-miR-18a-5p for sensitivity and hsa-miR-637 for resistance. Drug sensitivity was associated with hsa-let-7a-5p for Bortezomib, hsa-miR-135a-3p for JNJ-707 and hsa-miR-185-3p for Panobinostat. Drug resistance was associated with hsa-miR-182-5p for Veliparib and hsa-miR-629-5p for Tipifarnib. Pathway analysis for significant miRNAs was performed to reveal biological roles, aiding to find a potential mechanistic link for the observed associations with drug response. By doing so hsa-miR-187-5p was linked to the cell cycle G2-M checkpoint in line with this checkpoint being the target of taxanes. In conclusion, our study shows that miRNAs could potentially serve as biomarkers for intrinsic drug resistance and that pathway analyses can provide additional information in this context.
- Published
- 2019
45. Communication is key: Mother-offspring signaling can affect behavioral responses and offspring survival in feral horses (Equus caballus)
- Author
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Daniel I. Rubenstein and Cassandra M.V. Nuñez
- Subjects
0106 biological sciences ,Topography ,animal diseases ,Social Sciences ,Developmental Signaling ,Mother offspring ,01 natural sciences ,Developmental psychology ,Signal Initiation ,Sociology ,Cell Signaling ,Psychology ,Animal communication ,Mammals ,Islands ,Multidisciplinary ,Behavior, Animal ,Animal Behavior ,biology ,Physics ,Mechanisms of Signal Transduction ,05 social sciences ,Eukaryota ,Social Communication ,Foal ,Vertebrates ,Physical Sciences ,Medicine ,Female ,Research Article ,Signal Transduction ,Offspring ,Science ,Equines ,Affect (psychology) ,010603 evolutionary biology ,biology.animal ,Acoustic Signals ,Animals ,Juvenile ,0501 psychology and cognitive sciences ,Horses ,050102 behavioral science & comparative psychology ,Behavior ,Landforms ,Organisms ,Biology and Life Sciences ,Geomorphology ,Acoustics ,Cell Biology ,Bond formation ,biology.organism_classification ,Equus ,Communications ,Animal Communication ,Amniotes ,Earth Sciences ,Zoology - Abstract
Acoustic signaling plays an important role in mother-offspring recognition and subsequent bond-formation. It remains unclear, however, if mothers and offspring use acoustic signaling in the same ways and for the same reasons throughout the juvenile stage, particularly after mutual recognition has been adequately established. Moreover, despite its critical role in mother-offspring bond formation, research explicitly linking mother-infant communication strategies to offspring survival are lacking. We examined the communicative patterns of mothers and offspring in the feral horse (Equus caballus) to better understand 1) the nature of mother-offspring communication throughout the first year of development; 2) the function(s) of mother- vs. offspring-initiated communication and; 3) the importance of mare and foal communication to offspring survival. We found that 1) mares and foals differ in when and how they initiate communication; 2) the outcomes of mare- vs. foal-initiated communication events consistently differ; and 3) the communicative patterns between mares and their foals can be important for offspring survival to one year of age. Moreover, given the importance of maternal activity to offspring behavior and subsequent survival, we submit that our data are uniquely positioned to address the long-debated question: do the behaviors exhibited during the juvenile stage (by both mothers and their young) confer delayed or immediate benefits to offspring? In summary, we aimed to better understand 1) the dynamics of mother-offspring communication, 2) whether mother-offspring communicative patterns were important to offspring survival, and 3) the implications of our research regarding the function of the mammalian juvenile stage. Our results demonstrate that we have achieved those aims.
- Published
- 2020
46. An enhanced password authentication scheme for session initiation protocol with perfect forward secrecy
- Author
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Haseeb Ahmad, Shuming Qiu, Guoai Xu, and Yanhui Guo
- Subjects
computer.internet_protocol ,Computer science ,Health Smart Cards ,lcsh:Medicine ,Synthesis Phase ,Cryptography ,02 engineering and technology ,Internet security ,Polynomials ,Database and Informatics Methods ,Signal Initiation ,Forward secrecy ,0202 electrical engineering, electronic engineering, information engineering ,Password authentication protocol ,Cell Cycle and Cell Division ,Computer Networks ,Database Searching ,lcsh:Science ,Password ,Social Responsibility ,Multidisciplinary ,Communication ,Mechanisms of Signal Transduction ,Multimedia ,Cell Processes ,Physical Sciences ,The Internet ,Smart card ,Communications protocol ,Confidentiality ,Computer network ,Research Article ,Signal Transduction ,Information Systems ,Computer and Information Sciences ,Research and Analysis Methods ,Computer Security ,Session Initiation Protocol ,Authentication ,Internet ,business.industry ,lcsh:R ,Biology and Life Sciences ,020206 networking & telecommunications ,020207 software engineering ,Cell Biology ,Algebra ,lcsh:Q ,business ,computer ,Algebraic Geometry ,Mathematics ,Software - Abstract
The Session Initiation Protocol (SIP) is an extensive and esteemed communication protocol employed to regulate signaling as well as for controlling multimedia communication sessions. Recently, Kumari et al. proposed an improved smart card based authentication scheme for SIP based on Farash's scheme. Farash claimed that his protocol is resistant against various known attacks. But, we observe some accountable flaws in Farash's protocol. We point out that Farash's protocol is prone to key-compromise impersonation attack and is unable to provide pre-verification in the smart card, efficient password change and perfect forward secrecy. To overcome these limitations, in this paper we present an enhanced authentication mechanism based on Kumari et al.'s scheme. We prove that the proposed protocol not only overcomes the issues in Farash's scheme, but it can also resist against all known attacks. We also provide the security analysis of the proposed scheme with the help of widespread AVISPA (Automated Validation of Internet Security Protocols and Applications) software. At last, comparing with the earlier proposals in terms of security and efficiency, we conclude that the proposed protocol is efficient and more secure.
- Published
- 2018
47. Treatment effect of mTOR-inhibition on tissue composition of renal angiomyolipomas in tuberous sclerosis complex (TSC)
- Author
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Susanne Brakemeier, Klemens Budde, Bernd Hamm, Lisa C. Adams, Bianca Zukunft, Gerd Diederichs, Marcus R. Makowski, and Lars Vogt
- Subjects
Male ,Angiomyolipoma ,030232 urology & nephrology ,lcsh:Medicine ,Signal-To-Noise Ratio ,Pathology and Laboratory Medicine ,Biochemistry ,Vascular Medicine ,Diagnostic Radiology ,Fats ,Tuberous sclerosis ,0302 clinical medicine ,Signal Initiation ,Cell Signaling ,Tuberous Sclerosis ,Medicine and Health Sciences ,lcsh:Science ,Multidisciplinary ,medicine.diagnostic_test ,Radiology and Imaging ,TOR Serine-Threonine Kinases ,Mechanisms of Signal Transduction ,Middle Aged ,Magnetic Resonance Imaging ,Lipids ,Kidney Neoplasms ,Genetic Diseases ,Female ,Anatomy ,Tissue composition ,medicine.drug ,Research Article ,Signal Transduction ,Adult ,medicine.medical_specialty ,Signal Inhibition ,Imaging Techniques ,Urology ,Hemorrhage ,Research and Analysis Methods ,03 medical and health sciences ,Text mining ,Signs and Symptoms ,Diagnostic Medicine ,medicine ,Humans ,PI3K/AKT/mTOR pathway ,Clinical Genetics ,Sirolimus ,Everolimus ,business.industry ,lcsh:R ,Biology and Life Sciences ,Magnetic resonance imaging ,Kidneys ,Renal System ,Cell Biology ,medicine.disease ,lcsh:Q ,business ,030217 neurology & neurosurgery - Abstract
Purpose Tuberous sclerosis complex (TSC)-associated renal angiomyolipoma (AML) have a high lifetime risk of acute bleeding. MTOR-inhibitors are a promising novel treatment for TSC-AML, however adequate response to therapy can be difficult to assess. Early changes in MRI signal may serve as a novel early indicator for a satisfactory response to mTOR-inhibitor therapy of AML. Materials and methods Thirty-eight patients with the definite diagnosis of tuberous sclerosis receiving everolimus therapy and n = 19 patients without specific therapy were included. 1.5 Tesla MRI was performed including sequences with a selective fat suppression. Patients were investigated prior to the initiation of therapy (baseline) and after 0.05) and size (p>0.05) were measured in the control group. Conclusion mTOR inhibitor therapy in TSC patients results in an early and pronounced fatty transformation of AMLs on MRI. Fatty transformation could represent a novel early indicator of response to therapy in this patient collective.
- Published
- 2017
48. Data-derived modeling characterizes plasticity of MAPK signaling in melanoma
- Author
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Christine S. Falk, Stefan Massen, Roland Eils, Nathan R. Brady, and Marti Bernardo-Faura
- Subjects
Melanomas ,Cell signaling ,Somatic cell ,Cancer Treatment ,Signal transduction ,Crosstalk (Biology) ,Signal Initiation ,Medicine and Health Sciences ,Membrane Receptor Signaling ,Phosphorylation ,Melanoma ,lcsh:QH301-705.5 ,Genetics ,Network architecture ,Ecology ,Kinase ,Systems Biology ,Mechanisms of Signal Transduction ,Signaling cascades ,Hormone Receptor Signaling ,Oncology ,Computational Theory and Mathematics ,Cell Processes ,Modeling and Simulation ,Network Analysis ,Research Article ,Cell biology ,Computer and Information Sciences ,MAPK signaling cascades ,Feedback Regulation ,MAP Kinase Signaling System ,Phosphoinositide Signal Transduction ,Computational biology ,Biology ,Plasticity ,Models, Biological ,Cell Growth ,Cellular and Molecular Neuroscience ,Fuzzy Logic ,Cancer Genetics ,medicine ,Humans ,Theoretical Biology ,Molecular Biology ,Transcription factor ,Ecology, Evolution, Behavior and Systematics ,Biology and life sciences ,Computational Biology ,Cancers and Neoplasms ,medicine.disease ,Computing Methods ,Signaling Networks ,lcsh:Biology (General) ,Genetics of Disease - Abstract
The majority of melanomas have been shown to harbor somatic mutations in the RAS-RAF-MEK-MAPK and PI3K-AKT pathways, which play a major role in regulation of proliferation and survival. The prevalence of these mutations makes these kinase signal transduction pathways an attractive target for cancer therapy. However, tumors have generally shown adaptive resistance to treatment. This adaptation is achieved in melanoma through its ability to undergo neovascularization, migration and rearrangement of signaling pathways. To understand the dynamic, nonlinear behavior of signaling pathways in cancer, several computational modeling approaches have been suggested. Most of those models require that the pathway topology remains constant over the entire observation period. However, changes in topology might underlie adaptive behavior to drug treatment. To study signaling rearrangements, here we present a new approach based on Fuzzy Logic (FL) that predicts changes in network architecture over time. This adaptive modeling approach was used to investigate pathway dynamics in a newly acquired experimental dataset describing total and phosphorylated protein signaling over four days in A375 melanoma cell line exposed to different kinase inhibitors. First, a generalized strategy was established to implement a parameter-reduced FL model encoding non-linear activity of a signaling network in response to perturbation. Next, a literature-based topology was generated and parameters of the FL model were derived from the full experimental dataset. Subsequently, the temporal evolution of model performance was evaluated by leaving time-defined data points out of training. Emerging discrepancies between model predictions and experimental data at specific time points allowed the characterization of potential network rearrangement. We demonstrate that this adaptive FL modeling approach helps to enhance our mechanistic understanding of the molecular plasticity of melanoma., Author Summary Signal transduction pathways can be described as static routes, transmitting extrinsic signals to the nucleus to induce a transcriptional response. In contrast to this reductionist view, the emerging paradigm is that signaling networks undergo dynamic crosstalk, both in disease and physiological conditions. To understand complex pathway behavior, it is necessary to develop methods to identify pathway interactions that are active as a consequence of stimuli and, importantly, to describe their evolution in time. To that end, we developed a method relying on prior knowledge networks in order to predict signaling crosstalk evolution, in response to perturbation and over time. The challenge we addressed was to establish a method dependent on information related to the topology of reported interactions, and not their mechanistic characteristics, and at the same time complex enough to reproduce the behavior of the signaling intermediates. The work presented here demonstrates that such an approach can be used to predict mechanisms that melanoma uses to rearrange its signaling and maintain its abnormal proliferation upon treatment.
- Published
- 2014
49. Genomic and Genotoxic Responses to Controlled Weathered-Oil Exposures Confirm and Extend Field Studies on Impacts of the Deepwater Horizon Oil Spill on Native Killifish
- Author
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Scott Miles, Whitney Pilcher, Gregory D. Mayer, Andrew Whitehead, and Song Tang
- Subjects
Gulf killifish ,lcsh:Medicine ,Gene Expression ,Toxicology ,Ecotoxicology ,Transcriptome ,Disasters ,Signal Initiation ,Cell Signaling ,Fundulidae ,Molecular Cell Biology ,Petroleum Pollution ,Killifish ,lcsh:Science ,Oil toxicity ,Genetics ,Regulation of gene expression ,Multidisciplinary ,biology ,Mechanisms of Signal Transduction ,Genomics ,Fundulus ,Functional Genomics ,Petroleum ,Organ Specificity ,Transcriptome Analysis ,Research Article ,Signal Transduction ,DNA damage ,Genetic Toxicology ,Toxic Agents ,Zoology ,Animals ,Dose-Response Relationship, Drug ,Mutagenicity Tests ,lcsh:R ,Ecology and Environmental Sciences ,Biology and Life Sciences ,Computational Biology ,Cell Biology ,biology.organism_classification ,Genome Analysis ,13. Climate action ,lcsh:Q ,Genome Expression Analysis ,Environmental Protection ,DNA Damage - Abstract
To understand the ecotoxicological impacts of the Deepwater Horizon oil spill, field studies provide a context for ecological realism but laboratory-based studies offer power for connecting biological effects with specific causes. As a complement to field studies, we characterized genome-wide gene expression responses of Gulf killifish (Fundulus grandis) to oil-contaminated waters in controlled laboratory exposures. Transcriptional responses to the highest concentrations of oiled water in the laboratory were predictive of field-observed responses that coincided with the timing and location of major oiling. The transcriptional response to the low concentration (∼ 10-fold lower than the high concentration) was distinct from the high concentration and was not predictive of major oiling in the field. The high concentration response was characterized by activation of the molecular signaling pathway that facilitates oil metabolism and oil toxicity. The high concentration also induced DNA damage. The low concentration invoked expression of genes that may support a compensatory response, including genes associated with regulation of transcription, cell cycle progression, RNA processing, DNA damage, and apoptosis. We conclude that the gene expression response detected in the field was a robust indicator of exposure to the toxic components of contaminating oil, that animals in the field were exposed to relatively high concentrations that are especially damaging to early life stages, and that such exposures can damage DNA.
- Published
- 2014
50. Pannexin 1 Channels Play Essential Roles in Urothelial Mechanotransduction and Intercellular Signaling
- Author
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David C. Spray, Hiromitsu Negoro, Sylvia O. Suadicani, Mia M. Thi, Louis S. Liou, and Marcia Urban-Maldonado
- Subjects
Nucleotide Receptor Signaling ,lcsh:Medicine ,Stimulation ,Mechanotransduction, Cellular ,Epithelium ,Connexins ,Rats, Sprague-Dawley ,chemistry.chemical_compound ,Mice ,0302 clinical medicine ,Crosstalk (Biology) ,Signal Initiation ,Adenosine Triphosphate ,Cell Signaling ,Molecular Cell Biology ,Membrane Receptor Signaling ,Mechanotransduction ,lcsh:Science ,Mice, Knockout ,0303 health sciences ,Multidisciplinary ,Mechanisms of Signal Transduction ,Tumor Protein, Translationally-Controlled 1 ,Pannexin ,3. Good health ,Cell biology ,Mefloquine ,Anatomy ,Research Article ,Signal Transduction ,Autocrine Signaling ,Cell signaling ,Cell Physiology ,Nerve Tissue Proteins ,Biology ,03 medical and health sciences ,Paracrine signalling ,Antimalarials ,Animals ,Humans ,Channel blocker ,Calcium Signaling ,030304 developmental biology ,Apyrase ,lcsh:R ,Biology and Life Sciences ,Epithelial Cells ,Cell Biology ,Rats ,Biological Tissue ,chemistry ,lcsh:Q ,Calcium ,Receptors, Purinergic P2X7 ,Urothelium ,Adenosine triphosphate ,030217 neurology & neurosurgery ,HeLa Cells - Abstract
Urothelial cells respond to bladder distension with ATP release, and ATP signaling within the bladder and from the bladder to the CNS is essential for proper bladder function. In other cell types, pannexin 1 (Panx1) channels provide a pathway for mechanically-induced ATP efflux and for ATP-induced ATP release through interaction with P2X7 receptors (P2X7Rs). We report that Panx1 and P2X7R are functionally expressed in the bladder mucosa and in immortalized human urothelial cells (TRT-HU1), and participate in urothelial ATP release and signaling. ATP release from isolated rat bladders induced by distention was reduced by the Panx1 channel blocker mefloquine (MFQ) and was blunted in mice lacking Panx1 or P2X7R expression. Hypoosmotic shock induced YoPro dye uptake was inhibited by MFQ and the P2X7R blocker A438079 in TRT-HU1 cells, and was also blunted in primary urothelial cells derived from mice lacking Panx1 or P2X7R expression. Rinsing-induced mechanical stimulation of TRT-HU1 cells triggered ATP release, which was reduced by MFQ and potentiated in low divalent cation solution (LDPBS), a condition known to enhance P2X7R activation. ATP signaling evaluated as intercellular Ca2+ wave radius was significantly larger in LDPBS, reduced by MFQ and by apyrase (ATP scavenger). These findings indicate that Panx1 participates in urothelial mechanotransduction and signaling by providing a direct pathway for mechanically-induced ATP release and by functionally interacting with P2X7Rs.
- Published
- 2014
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