19 results on '"Moutasim KA"'
Search Results
2. Investigating the Mechanisms Underlying Tumour Immune Evasion in Head and Neck Squamous Cell Carcinoma
- Author
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Robinson, H, Fleming, JC, Thirdborough, S, Ottensmeier, C, Thomas, GJ, and Moutasim, KA
- Published
- 2019
3. Pro-migratory and TGF-β-activating functions of αvβ6 integrin in pancreatic cancer are differentially regulated via an Eps8-dependent GTPase switch
- Author
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Tod, J, Hanley, CJ, Morgan, MR, Rucka, M, Mellows, T, Lopez, M-A, Kiely, P, Moutasim, KA, Frampton, SJ, Sabnis, D, Fine, DR, Johnson, C, Marshall, JF, Scita, G, Jenei, V, and Thomas, GJ
- Abstract
The integrin αvβ6 is upregulated in numerous carcinomas, where expression commonly correlates with poor prognosis. αvβ6 promotes tumour invasion, partly through regulation of proteases and cell migration, and is also the principal mechanism by which epithelial cells activate TGF-β1; this latter function complicates therapeutic targeting of αvβ6, since TGF-β1 has both tumour-promoting and -suppressive effects. It is unclear how these different αvβ6 functions are linked; both require actin cytoskeletal reorganisation, and it is suggested that tractive forces generated during cell migration activate TGF-β1 by exerting mechanical tension on the ECM-bound latent complex. We examined the functional relationship between cell invasion and TGF-β1 activation in pancreatic ductal adenocarcinoma (PDAC) cells, and confirmed that both processes are αvβ6-dependent. Surprisingly, we found that cellular functions could be biased towards either motility or TGF-β1 activation depending on the presence or absence of epidermal growth factor receptor pathway substrate 8 (Eps8), a regulator of actin remodelling, endocytosis and GTPase activation. Similar to αvβ6, we found that Eps8 was upregulated in >70% of PDAC. In complex with Abi1/Sos1, Eps8 regulated αvβ6-dependent cell migration through activation of Rac1. Downregulation of Eps8, Sos1 or Rac1 suppressed cell movement, while simultaneously increasing αvβ6-dependent TGF-β1 activation. This latter effect was modulated through increased cell tension, regulated by Rho activation. Thus, the Eps8/Abi1/Sos1 tricomplex acts as a key molecular switch altering the balance between Rac1 and Rho activation; its presence or absence in PDAC cells modulates αvβ6-dependent functions, resulting in a pro-migratory (Rac1-dependent) or a pro-TGF-β1 activation (Rho-dependent) functional phenotype respectively.
- Published
- 2017
4. Orofacial presentations of sarcoidosis--a case series and review of the literature.
- Author
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Poate TW, Sharma R, Moutasim KA, Escudier MP, and Warnakulasuriya S
- Published
- 2008
- Full Text
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5. Evaluation of the quality of RNA extracted from archival FFPE glioblastoma and epilepsy surgical samples for gene expression assays.
- Author
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Haynes HR, Killick-Cole CL, Hares KM, Redondo J, Kemp KC, Moutasim KA, Faulkner C, Wilkins A, and Kurian KM
- Subjects
- Brain Neoplasms surgery, Case-Control Studies, Epilepsy, Temporal Lobe surgery, Glioblastoma surgery, Humans, Predictive Value of Tests, Quality Control, Reproducibility of Results, Time Factors, Tissue Fixation standards, Brain Neoplasms genetics, Epilepsy, Temporal Lobe genetics, Fixatives chemistry, Formaldehyde chemistry, Gene Expression Profiling standards, Glioblastoma genetics, Paraffin Embedding standards, RNA Stability, RNA, Neoplasm genetics, Tissue Fixation methods
- Abstract
Aims: Histopathological tissue samples are being increasingly used as sources of nucleic acids in molecular pathology translational research. This study investigated the suitability of glioblastoma and control central nervous system (CNS) formalin-fixed paraffin embedded (FFPE) tissue-derived RNA for gene expression analyses., Methods: Total RNA was extracted from control (temporal lobe resection tissue) and glioblastoma FFPE tissue samples. RNA purity (260/280 ratios) was determined and RNA integrity number (RIN) analysis was performed. RNA was subsequently used for RT-qPCR for two reference genes, 18S and GAPDH ., Results: Reference gene expression was equivalent between control and glioblastoma tissue when using RNA extracted from FFPE tissue, which has key implications for biological normalisation for CNS gene expression studies. There was a significant difference between the mean RIN values of control and glioblastoma FFPE tissue. There was no significant correlation between 260/280 or RIN values versus total RNA yield. The age of the tissue blocks did not influence RNA yield, fragmentation or purity. There was no significant correlation between RIN or 260/280 ratios and mean qPCR cycle threshold for either reference gene., Conclusions: This study showed that routinely available CNS FFPE tissue is suitable for RNA extraction and downstream gene expression studies, even after 60 months of storage. Substantial RNA fragmentation associated with glioblastoma and control FFPE tissue blocks did not preclude downstream RT-qPCR gene expression analyses. Cross validation with both archival and prospectively collated FFPE specimens is required to further demonstrate that CNS tissue blocks can be used in novel translational molecular biomarker studies., Competing Interests: Competing interests: None declared., (© Article author(s) (or their employer(s) unless otherwise stated in the text of the article) 2018. All rights reserved. No commercial use is permitted unless otherwise expressly granted.)
- Published
- 2018
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6. Nodular fasciitis of the temporomandibular joint: a case report.
- Author
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Jenkyn I, King A, Moutasim KA, and Sharma S
- Subjects
- Diagnosis, Differential, Diagnostic Imaging, Fasciitis genetics, Fasciitis surgery, Gene Rearrangement, Humans, In Situ Hybridization, Fluorescence, Magnetic Resonance Imaging methods, Mutation, Proto-Oncogene Proteins genetics, Temporomandibular Joint surgery, Ubiquitin Thiolesterase genetics, Fascia diagnostic imaging, Fascia pathology, Fasciitis diagnostic imaging, Fasciitis pathology, Temporomandibular Joint diagnostic imaging, Temporomandibular Joint pathology
- Abstract
Nodular fasciitis is a relatively rare benign lesion of the soft tissue, which often presents in the fascia or deep subcutaneous tissues. It most commonly presents in the upper extremities and trunk and the head and neck region, particularly in younger patients. Its pathogenesis is poorly understood and it is predominantly thought to be a reactive lesion, although some have suggested that it may be a benign neoplasm. Advances in molecular testing and imaging have greatly assisted diagnosis. We discuss the benefits of ubiquitin-specific protease 6 (USP6) gene rearrangement testing and magnetic resonance imaging (MRI) to aid this uncommon diagnosis., (Copyright © 2017 The British Association of Oral and Maxillofacial Surgeons. Published by Elsevier Ltd. All rights reserved.)
- Published
- 2018
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7. Targeting the Myofibroblastic Cancer-Associated Fibroblast Phenotype Through Inhibition of NOX4.
- Author
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Hanley CJ, Mellone M, Ford K, Thirdborough SM, Mellows T, Frampton SJ, Smith DM, Harden E, Szyndralewiez C, Bullock M, Noble F, Moutasim KA, King EV, Vijayanand P, Mirnezami AH, Underwood TJ, Ottensmeier CH, and Thomas GJ
- Subjects
- Actins analysis, Adenocarcinoma chemistry, Adenocarcinoma genetics, Adult, Aged, Aged, 80 and over, Animals, Cancer-Associated Fibroblasts chemistry, Cancer-Associated Fibroblasts physiology, Carcinoma, Non-Small-Cell Lung chemistry, Carcinoma, Non-Small-Cell Lung genetics, Carcinoma, Squamous Cell chemistry, Carcinoma, Squamous Cell genetics, Cell Count, Cell Transdifferentiation drug effects, Cell Transdifferentiation genetics, Colorectal Neoplasms pathology, Disease Progression, Esophageal Neoplasms chemistry, Esophageal Neoplasms genetics, Female, Head and Neck Neoplasms chemistry, Head and Neck Neoplasms drug therapy, Head and Neck Neoplasms genetics, Humans, Lung Neoplasms chemistry, Lung Neoplasms genetics, Male, Mice, Middle Aged, Mouth Neoplasms pathology, Myofibroblasts chemistry, NADPH Oxidase 4, NADPH Oxidases analysis, NADPH Oxidases genetics, Neoplasm Transplantation, Oropharyngeal Neoplasms pathology, Phenotype, Pyrazoles therapeutic use, Pyrazolones, Pyridines therapeutic use, Pyridones, RNA Interference, Reactive Oxygen Species metabolism, Survival Rate, Up-Regulation, Adenocarcinoma drug therapy, Cancer-Associated Fibroblasts pathology, Carcinoma, Non-Small-Cell Lung drug therapy, Carcinoma, Squamous Cell drug therapy, Colorectal Neoplasms chemistry, Esophageal Neoplasms drug therapy, Lung Neoplasms drug therapy, Mouth Neoplasms chemistry, Myofibroblasts pathology, NADPH Oxidases antagonists & inhibitors, Oropharyngeal Neoplasms chemistry
- Abstract
Background: Cancer-associated fibroblasts (CAFs) are tumor-promoting and correlate with poor survival in many cancers, which has led to their emergence as potential therapeutic targets. However, effective methods to manipulate these cells clinically have yet to be developed., Methods: CAF accumulation and prognostic significance in head and neck cancer (oral, n = 260; oropharyngeal, n = 271), and colorectal cancer (n = 56) was analyzed using immunohistochemistry. Mechanisms regulating fibroblast-to-myofibroblast transdifferentiation were investigated in vitro using RNA interference/pharmacological inhibitors followed by polymerase chain reaction (PCR), immunoblotting, immunofluorescence, and functional assays. RNA sequencing/bioinformatics and immunohistochemistry were used to analyze NAD(P)H Oxidase-4 (NOX4) expression in different human tumors. NOX4's role in CAF-mediated tumor progression was assessed in vitro, using CAFs from multiple tissues in Transwell and organotypic culture assays, and in vivo, using xenograft (n = 9-15 per group) and isograft (n = 6 per group) tumor models. All statistical tests were two-sided., Results: Patients with moderate/high levels of myofibroblastic-CAF had a statistically significant decrease in cancer-specific survival rates in each cancer type analyzed (hazard ratios [HRs] = 1.69-7.25, 95% confidence intervals [CIs] = 1.11 to 31.30, log-rank P ≤ .01). Fibroblast-to-myofibroblast transdifferentiation was dependent on a delayed phase of intracellular reactive oxygen species, generated by NOX4, across different anatomical sites and differentiation stimuli. A statistically significant upregulation of NOX4 expression was found in multiple human cancers (P < .001), strongly correlating with myofibroblastic-CAFs (r = 0.65-0.91, adjusted P < .001). Genetic/pharmacological inhibition of NOX4 was found to revert the myofibroblastic-CAF phenotype ex vivo (54.3% decrease in α-smooth muscle actin [α-SMA], 95% CI = 10.6% to 80.9%, P = .009), prevent myofibroblastic-CAF accumulation in vivo (53.2%-79.0% decrease in α-SMA across different models, P ≤ .02) and slow tumor growth (30.6%-64.0% decrease across different models, P ≤ .04)., Conclusions: These data suggest that pharmacological inhibition of NOX4 may have broad applicability for stromal targeting across cancer types., (© The Author 2017. Published by Oxford University Press.)
- Published
- 2018
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8. Evaluating the effect of immune cells on the outcome of patients with mesothelioma.
- Author
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Chee SJ, Lopez M, Mellows T, Gankande S, Moutasim KA, Harris S, Clarke J, Vijayanand P, Thomas GJ, and Ottensmeier CH
- Subjects
- Adult, Aged, Aged, 80 and over, Cohort Studies, Female, Follow-Up Studies, Humans, Male, Mesothelioma pathology, Middle Aged, Neoplasm Staging, Prognosis, Survival Rate, B-Lymphocytes immunology, Killer Cells, Natural immunology, Lymphocytes, Tumor-Infiltrating immunology, Mesothelioma immunology, Mesothelioma mortality, Tumor Microenvironment immunology
- Abstract
Background: We systematically assessed the prognostic and predictive value of infiltrating adaptive and innate immune cells in a large cohort of patients with advanced mesothelioma., Methods: A tissue microarray from 302 samples was constructed. Markers of adaptive immune response in T-cells (CD8
+ , FOXP3+ , CD4+ , CD45RO+ , CD3+ ) and B-cells (CD20+ ), and of innate immune response; neutrophils (NP57+ ), natural killer cells (CD56+ ) and macrophages (CD68+ ) were evaluated., Results: We found that in the epithelioid tumours, high CD4+ and CD20+ counts, and low FOXP3+ , CD68+ and NP57+ counts linked to better outcome. In the non-epithelioid group low CD8+ and low FOXP3+ counts were beneficial.On multivariate analysis low FOXP3+ remained independently associated with survival in both groups. In the epithelioid group additionally high CD4+ , high CD20+ , and low NP57+ counts were prognostic., Conclusions: Our data demonstrate for the first time, in predominately advanced disease, the association of key markers of adaptive and innate immunity with survival and the differential effect of histology. A better understanding of the immunological drivers of the different subtypes of mesothelioma will assist prognostication and disease-specific clinical decision-making.- Published
- 2017
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9. Pro-migratory and TGF-β-activating functions of αvβ6 integrin in pancreatic cancer are differentially regulated via an Eps8-dependent GTPase switch.
- Author
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Tod J, Hanley CJ, Morgan MR, Rucka M, Mellows T, Lopez MA, Kiely P, Moutasim KA, Frampton SJ, Sabnis D, Fine DR, Johnson C, Marshall JF, Scita G, Jenei V, and Thomas GJ
- Subjects
- Adaptor Proteins, Signal Transducing genetics, Antigens, Neoplasm genetics, Carcinoma, Pancreatic Ductal genetics, Carcinoma, Pancreatic Ductal pathology, Cell Line, Tumor, Coculture Techniques, Cytoskeletal Proteins genetics, Cytoskeletal Proteins metabolism, Enzyme Inhibitors pharmacology, Humans, Integrins genetics, Neoplasm Invasiveness, Pancreatic Neoplasms genetics, Pancreatic Neoplasms pathology, Phenotype, RNA Interference, SOS1 Protein genetics, SOS1 Protein metabolism, Signal Transduction, Stromal Cells enzymology, Stromal Cells pathology, Transfection, Tumor Microenvironment, rac1 GTP-Binding Protein genetics, rho GTP-Binding Proteins antagonists & inhibitors, Adaptor Proteins, Signal Transducing metabolism, Antigens, Neoplasm metabolism, Carcinoma, Pancreatic Ductal enzymology, Cell Movement drug effects, Integrins metabolism, Pancreatic Neoplasms enzymology, Transforming Growth Factor beta1 metabolism, rac1 GTP-Binding Protein metabolism, rho GTP-Binding Proteins metabolism
- Abstract
The integrin αvβ6 is up-regulated in numerous carcinomas, where expression commonly correlates with poor prognosis. αvβ6 promotes tumour invasion, partly through regulation of proteases and cell migration, and is also the principal mechanism by which epithelial cells activate TGF-β1; this latter function complicates therapeutic targeting of αvβ6, since TGF-β1 has both tumour-promoting and -suppressive effects. It is unclear how these different αvβ6 functions are linked; both require actin cytoskeletal reorganization, and it is suggested that tractive forces generated during cell migration activate TGF-β1 by exerting mechanical tension on the ECM-bound latent complex. We examined the functional relationship between cell invasion and TGF-β1 activation in pancreatic ductal adenocarcinoma (PDAC) cells, and confirmed that both processes are αvβ6-dependent. Surprisingly, we found that cellular functions could be biased towards either motility or TGF-β1 activation depending on the presence or absence of epidermal growth factor receptor pathway substrate 8 (Eps8), a regulator of actin remodelling, endocytosis, and GTPase activation. Similar to αvβ6, we found that Eps8 was up-regulated in >70% of PDACs. In complex with Abi1/Sos1, Eps8 regulated αvβ6-dependent cell migration through activation of Rac1. Down-regulation of Eps8, Sos1 or Rac1 suppressed cell movement, while simultaneously increasing αvβ6-dependent TGF-β1 activation. This latter effect was modulated through increased cell tension, regulated by Rho activation. Thus, the Eps8/Abi1/Sos1 tricomplex acts as a key molecular switch altering the balance between Rac1 and Rho activation; its presence or absence in PDAC cells modulates αvβ6-dependent functions, resulting in a pro-migratory (Rac1-dependent) or a pro-TGF-β1 activation (Rho-dependent) functional phenotype, respectively. © 2017 The Authors. The Journal of Pathology published by John Wiley & Sons Ltd on behalf of Pathological Society of Great Britain and Ireland., (© 2017 The Authors. The Journal of Pathology published by John Wiley & Sons Ltd on behalf of Pathological Society of Great Britain and Ireland.)
- Published
- 2017
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10. Crowdsourcing for translational research: analysis of biomarker expression using cancer microarrays.
- Author
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Lawson J, Robinson-Vyas RJ, McQuillan JP, Paterson A, Christie S, Kidza-Griffiths M, McDuffus LA, Moutasim KA, Shaw EC, Kiltie AE, Howat WJ, Hanby AM, Thomas GJ, and Smittenaar P
- Subjects
- Data Interpretation, Statistical, Humans, Image Processing, Computer-Assisted methods, Immunohistochemistry, Patient Selection, Biomarkers, Tumor metabolism, Crowdsourcing methods, Neoplasms metabolism, Tissue Array Analysis, Translational Research, Biomedical methods
- Abstract
Background: Academic pathology suffers from an acute and growing lack of workforce resource. This especially impacts on translational elements of clinical trials, which can require detailed analysis of thousands of tissue samples. We tested whether crowdsourcing - enlisting help from the public - is a sufficiently accurate method to score such samples., Methods: We developed a novel online interface to train and test lay participants on cancer detection and immunohistochemistry scoring in tissue microarrays. Lay participants initially performed cancer detection on lung cancer images stained for CD8, and we measured how extending a basic tutorial by annotated example images and feedback-based training affected cancer detection accuracy. We then applied this tutorial to additional cancer types and immunohistochemistry markers - bladder/ki67, lung/EGFR, and oesophageal/CD8 - to establish accuracy compared with experts. Using this optimised tutorial, we then tested lay participants' accuracy on immunohistochemistry scoring of lung/EGFR and bladder/p53 samples., Results: We observed that for cancer detection, annotated example images and feedback-based training both improved accuracy compared with a basic tutorial only. Using this optimised tutorial, we demonstrate highly accurate (>0.90 area under curve) detection of cancer in samples stained with nuclear, cytoplasmic and membrane cell markers. We also observed high Spearman correlations between lay participants and experts for immunohistochemistry scoring (0.91 (0.78, 0.96) and 0.97 (0.91, 0.99) for lung/EGFR and bladder/p53 samples, respectively)., Conclusions: These results establish crowdsourcing as a promising method to screen large data sets for biomarkers in cancer pathology research across a range of cancers and immunohistochemical stains.
- Published
- 2017
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11. Induction of fibroblast senescence generates a non-fibrogenic myofibroblast phenotype that differentially impacts on cancer prognosis.
- Author
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Mellone M, Hanley CJ, Thirdborough S, Mellows T, Garcia E, Woo J, Tod J, Frampton S, Jenei V, Moutasim KA, Kabir TD, Brennan PA, Venturi G, Ford K, Herranz N, Lim KP, Clarke J, Lambert DW, Prime SS, Underwood TJ, Vijayanand P, Eliceiri KW, Woelk C, King EV, Gil J, Ottensmeier CH, and Thomas GJ
- Subjects
- Animals, Cell Line, Tumor, Extracellular Matrix metabolism, Fibroblasts metabolism, Humans, Mice, Myofibroblasts metabolism, Neoplasms metabolism, Phenotype, Prognosis, Signal Transduction physiology, Transforming Growth Factor beta1 metabolism, Cell Differentiation physiology, Cellular Senescence physiology, Fibroblasts pathology, Myofibroblasts pathology, Neoplasms pathology
- Abstract
Cancer-associated fibroblasts (CAF) remain a poorly characterized, heterogeneous cell population. Here we characterized two previously described tumor-promoting CAF sub-types, smooth muscle actin (SMA)-positive myofibroblasts and senescent fibroblasts, identifying a novel link between the two. Analysis of CAF cultured ex vivo , showed that senescent CAF are predominantly SMA-positive; this was confirmed by immunochemistry in head & neck (HNSCC) and esophageal (EAC) cancers. In vitro , we found that fibroblasts induced to senesce develop molecular, ultrastructural and contractile features typical of myofibroblasts and this is dependent on canonical TGF-β signaling. Similar to TGF-β1-generated myofibroblasts, these cells secrete soluble factors that promote tumor cell motility. However, RNA-sequencing revealed significant transcriptomic differences between the two SMA-positive CAF groups, particularly in genes associated with extracellular matrix (ECM) deposition and organization, which differentially promote tumor cell invasion. Notably, second harmonic generation imaging and bioinformatic analysis of SMA-positive human HNSCC and EAC showed that collagen fiber organization correlates with poor prognosis, indicating that heterogeneity within the SMA-positive CAF population differentially impacts on survival. These results show that non-fibrogenic, SMA-positive myofibroblasts can be directly generated through induction of fibroblast senescence and suggest that senescence and myofibroblast differentiation are closely linked processes.
- Published
- 2016
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12. Suppression of Hedgehog signalling promotes pro-tumourigenic integrin expression and function.
- Author
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Moutasim KA, Mellows T, Mellone M, Lopez MA, Tod J, Kiely PC, Sapienza K, Greco A, Neill GW, Violette S, Weinreb PH, Marshall JF, Ottensmeier CH, Sayan AE, Jenei V, and Thomas GJ
- Subjects
- Antigens, Neoplasm genetics, Carcinoma, Basal Cell genetics, Carcinoma, Basal Cell pathology, Cell Line, Cell Movement, Cell Transformation, Neoplastic genetics, Cell Transformation, Neoplastic pathology, Coculture Techniques, Down-Regulation, Fibroblasts metabolism, Fibroblasts pathology, Humans, Integrins genetics, Keratinocytes metabolism, Keratinocytes pathology, Pancreatic Neoplasms genetics, Pancreatic Neoplasms pathology, RNA Interference, Skin Neoplasms genetics, Skin Neoplasms pathology, Smad2 Protein metabolism, Smad3 Protein metabolism, Transcription Factors genetics, Transfection, Transforming Growth Factor beta1 metabolism, Zinc Finger Protein GLI1, Antigens, Neoplasm metabolism, Carcinoma, Basal Cell metabolism, Cell Transformation, Neoplastic metabolism, Hedgehog Proteins metabolism, Integrins metabolism, Pancreatic Neoplasms metabolism, Signal Transduction, Skin Neoplasms metabolism, Transcription Factors metabolism
- Abstract
Aberrant Hedgehog (Hh) signalling has been reported in a number of malignancies, particularly basal cell carcinoma (BCC) of the skin. Clinical trials of Hh inhibitors are underway in many cancers, and these have produced significant clinical benefit in BCC patients, although regrowth of new, or clinically aggressive, variants, as well as development of secondary malignancies, has been reported. αvβ6 integrin is expressed in many cancers, where it has been shown to correlate with an aggressive tumour phenotype and poor prognosis. We have previously reported αvβ6 up-regulation in aggressive, morphoeic BCC variants, where it modulates the stromal response and induces invasion. To examine a possible link between Hh and αvβ6 function, we generated BCC models, overexpressing Gli1 in immortalized keratinocytes (NTert1, HaCaT). Unexpectedly, we found that suppressing Gli1 significantly increased αvβ6 expression. This promoted tumour cell motility and also stromal myofibroblast differentiation through integrin-dependent TGF-β1 activation. Gli1 inhibited αvβ6 expression by suppressing TGF-β1-induced Smad2/3 activation, blocking a positive feedback loop maintaining high αvβ6 levels. A similar mechanism was observed in AsPC1 pancreatic cancer cells expressing endogenous Gli1, suggesting a common mechanism across tumour types. In vitro findings were supported using human clinical samples, where we showed an inverse correlation between αvβ6 and Gli1 expression in different BCC subtypes and pancreatic cancers. In summary, we show that expression of Gli1 and αvβ6 inversely correlates in tumours in vivo, and Hh targeting up-regulates TGF-β1/Smad2/3-dependent αvβ6 expression, promoting pro-tumourigenic cell functions in vitro. These results have potential clinical significance, given the reported recurrence of BCC variants and secondary malignancies in patients treated by Hh targeting., (Copyright © 2014 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.)
- Published
- 2014
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13. In vitro effect of bisphosphonates on oral keratinocytes and fibroblasts.
- Author
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McLeod NM, Moutasim KA, Brennan PA, Thomas G, and Jenei V
- Subjects
- Alendronate adverse effects, Bone Density Conservation Agents adverse effects, Cell Proliferation drug effects, Cells, Cultured, Clodronic Acid adverse effects, Coculture Techniques, Fetus, Foreskin cytology, Foreskin drug effects, Humans, Imidazoles adverse effects, Male, Mouth Mucosa cytology, Mouth Mucosa drug effects, Zoledronic Acid, Cell Movement drug effects, Diphosphonates adverse effects, Fibroblasts drug effects, Keratinocytes drug effects
- Abstract
Purpose: Osteonecrosis of the jaws is a potential complication of bisphosphonate (BP) therapy. The underlying mechanisms remain unclear. Although most research has concentrated on the effects of BPs on osteoclast and osteoblast functions, the disease is diagnosed and classified based on of mucosal breakdown, suggesting that oral soft tissues may be involved in its pathogenesis. The aim of this study was to determine the effect of 3 different BP drugs (alendronate, zoledronate, and clodronate) on the function of oral keratinocytes and fibroblasts., Materials and Methods: Human oral keratinocytes (OKF6) and fetal foreskin fibroblasts (HFFF2) were exposed to each drug at several concentrations and the effect on cell proliferation was assessed by counting the viable cells after different lengths of treatment. The effect on cell migration was examined using Transwell migration assays. An organotypic coculture model using keratinocytes and fibroblasts, which recapitulated the morphology of the oral mucosa, was used to assess the effect of the drugs on epithelial stratification and differentiation., Results: The 3 BPs affected the viability and proliferation of OKF6 and HFFF2 cells at concentrations in keeping with their known relative in vitro potencies. There was no effect on cell migration or tissue architecture in organotypic cultures at subtoxic concentrations., Conclusion: The lack of effect of these drugs on cell migration below concentrations known to affect cell viability suggests that BP-related osteonecrosis is not caused through suppression of keratinocyte or fibroblast motility., (Copyright © 2014 American Association of Oral and Maxillofacial Surgeons. Published by Elsevier Inc. All rights reserved.)
- Published
- 2014
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14. Stromal features are predictive of disease mortality in oral cancer patients.
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Marsh D, Suchak K, Moutasim KA, Vallath S, Hopper C, Jerjes W, Upile T, Kalavrezos N, Violette SM, Weinreb PH, Chester KA, Chana JS, Marshall JF, Hart IR, Hackshaw AK, Piper K, and Thomas GJ
- Subjects
- Actins metabolism, Aged, Biomarkers, Tumor metabolism, Carcinoma, Squamous Cell diagnosis, Carcinoma, Squamous Cell secondary, Carcinoma, Squamous Cell therapy, Epidemiologic Methods, Female, Humans, Male, Middle Aged, Mouth Neoplasms diagnosis, Mouth Neoplasms therapy, Myofibroblasts physiology, Neoplasm Invasiveness, Neoplasm Proteins metabolism, Neoplasm Staging, Prognosis, Stromal Cells metabolism, Carcinoma, Squamous Cell pathology, Mouth Neoplasms pathology, Stromal Cells pathology
- Abstract
Worldwide, approximately 405 000 cases of oral cancer (OSCC) are diagnosed each year, with a rising incidence in many countries. Despite advances in surgery and radiotherapy, which remain the standard treatment options, the mortality rate has remained largely unchanged for decades, with a 5-year survival rate of around 50%. OSCC is a heterogeneous disease, staged currently using the TNM classification, supplemented with pathological information from the primary tumour and loco-regional lymph nodes. Although patients with advanced disease show reduced survival, there is no single pathological or molecular feature that identifies aggressive, early-stage tumours. We retrospectively analysed 282 OSCC patients for disease mortality, related to clinical, pathological, and molecular features based on our previous functional studies [EGFR, αvβ6 integrin, smooth muscle actin (SMA), p53, p16, EP4]. We found that the strongest independent risk factor of early OSCC death was a feature of stroma rather than tumour cells. After adjusting for all factors, high stromal SMA expression, indicating myofibroblast transdifferentiation, produced the highest hazard ratio (3.06, 95% CI 1.65-5.66) and likelihood ratio (3.6; detection rate: false positive rate) of any feature examined, and was strongly associated with mortality, regardless of disease stage. Functional assays showed that OSCC cells can modulate myofibroblast transdifferentiation through αvβ6-dependent TGF-β1 activation and that myofibroblasts promote OSCC invasion. Finally, we developed a prognostic model using Cox regression with backward elimination; only SMA expression, metastasis, cohesion, and age were significant. This model was independently validated on a patient subset (detection rate 70%; false positive rate 20%; ROC analysis 77%, p < 0.001). Our study highlights the limited prognostic value of TNM staging and suggests that an SMA-positive, myofibroblastic stroma is the strongest predictor of OSCC mortality. Whether used independently or as part of a prognostic model, SMA identifies a significant group of patients with aggressive tumours, regardless of disease stage., (Copyright © 2011 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.)
- Published
- 2011
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15. Betel-derived alkaloid up-regulates keratinocyte alphavbeta6 integrin expression and promotes oral submucous fibrosis.
- Author
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Moutasim KA, Jenei V, Sapienza K, Marsh D, Weinreb PH, Violette SM, Lewis MP, Marshall JF, Fortune F, Tilakaratne WM, Hart IR, and Thomas GJ
- Subjects
- Actins metabolism, Antigens, Neoplasm genetics, Carcinoma, Squamous Cell metabolism, Carcinoma, Squamous Cell pathology, Cell Differentiation drug effects, Cell Movement drug effects, Cells, Cultured, Coculture Techniques, Humans, Integrins genetics, Keratinocytes metabolism, Mouth Neoplasms metabolism, Mouth Neoplasms pathology, Myofibroblasts cytology, Myofibroblasts drug effects, Oral Submucous Fibrosis pathology, Precancerous Conditions metabolism, Precancerous Conditions pathology, Reverse Transcriptase Polymerase Chain Reaction methods, Transforming Growth Factor beta1 metabolism, Up-Regulation drug effects, Antigens, Neoplasm biosynthesis, Areca chemistry, Arecoline pharmacology, Integrins biosynthesis, Keratinocytes drug effects, Oral Submucous Fibrosis metabolism
- Abstract
Oral submucous fibrosis (OSF) is a premalignant, fibrosing disorder of the mouth, pharynx, and oesophagus, with a malignant transformation rate of 7-13%. OSF is strongly associated with areca (betel) nut chewing and worldwide, over 5 million people are affected. As αvβ6 integrin is capable of promoting both tissue fibrosis and carcinoma invasion, we examined its expression in fibroepithelial hyperplasia and OSF. αvβ6 was markedly up-regulated in OSF, with high expression detected in 22 of 41 cases (p < 0.001). We investigated the functional role of αvβ6 using oral keratinocyte-derived cells genetically modified to express high αvβ6 (VB6), and also NTERT-immortalized oral keratinocytes, which express low αvβ6 (OKF6/TERT-1). VB6 cells showed significant αvβ6-dependent activation of TGF-β1, which induced transdifferentiation of oral fibroblasts into myofibroblasts and resulted in up-regulation of genes associated with tissue fibrosis. These experimental in vitro findings were confirmed using human clinical samples, where we showed that the stroma of OSF contained myofibroblasts and that TGF-β1-dependent Smad signalling was detectable both in keratinocytes and in myofibroblasts. We also found that arecoline, the major alkaloid of areca nuts, up-regulated keratinocyte αvβ6 expression. This was modulated through the M(4) muscarinic acetylcholine receptor and was suppressed by the M(4) antagonist, tropicamide. Arecoline-dependent αvβ6 up-regulation promoted keratinocyte migration and induced invasion, raising the possibility that this mechanism may support malignant transformation. Over 80% of OSF-related oral cancers examined had moderate/high αvβ6 expression. These data suggest that the pathogenesis of OSF may be epithelial-driven and involve arecoline-dependent up-regulation of αvβ6 integrin., (Copyright © 2010 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.)
- Published
- 2011
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16. Cell migration and invasion assays.
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Moutasim KA, Nystrom ML, and Thomas GJ
- Subjects
- Animals, Cell Culture Techniques, Cell Line, Tumor, Cell Movement, Cytological Techniques methods, Neoplasm Invasiveness
- Abstract
A number of in vitro assays have been developed to study tumor cell motility. Historically, assays have been mainly monocellular, where carcinoma cells are studied in isolation. Scratch assays can be used to study the collective and directional movement of populations of cells, whereas two chamber assays lend themselves to the analysis of chemotactic/haptotactic migration and cell invasion. However, an inherent disadvantage of these assays is that they grossly oversimplify the complex process of invasion, lacking the tumor structural architecture and stromal components. Organotypic assays, where tumor cells are grown at an air/liquid interface on gels populated with stromal cells, are a more physiologically relevant method for studying 3-dimensional tumor invasion.
- Published
- 2011
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17. A comparison of primary oesophageal squamous epithelial cells with HET-1A in organotypic culture.
- Author
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Underwood TJ, Derouet MF, White MJ, Noble F, Moutasim KA, Smith E, Drew PA, Thomas GJ, Primrose JN, and Blaydes JP
- Subjects
- Adenocarcinoma pathology, Antigens, CD biosynthesis, Cadherins biosynthesis, Casein Kinases biosynthesis, Coculture Techniques, Epithelial Cells metabolism, Esophagus cytology, Humans, Membrane Proteins biosynthesis, Vimentin biosynthesis, Barrett Esophagus pathology, Carcinoma, Squamous Cell pathology, Epithelial Cells pathology, Esophageal Neoplasms pathology
- Abstract
Background Information: Carcinoma of the oesophagus is the sixth leading cause of cancer death in the western world and is associated with a 5-year survival of less than 15%. Recent evidence suggests that stromal-epithelial interactions are fundamental in carcinogenesis. The advent of co-culture techniques permits the investigation of cross-talk between the stroma and epithelium in a physiological setting. We have characterized a histologically representative oesophageal organotypic model and have used it to compare the most commonly used squamous oesophageal cell line, HET-1A, with primary oesophageal squamous cells for use in studies of the oesophageal epithelium in vitro., Results: When grown in an organotypic culture with normal fibroblasts, the oesophageal carcinoma cell lines OE21 (squamous) and OE19 (adenocarcinoma) morphologically resembled the tumour of origin with evidence of stromal invasion and mucus production, respectively. However, HET-1A cells, which were derived from normal squamous oesophageal cells, appeared dysplastic and failed to display evidence of squamous differentiation. By comparison with primary oesophageal epithelial cells, the HET-1A cells were highly proliferative and did not express the epithelial markers E-cadherin or CK5/6 (casein kinase 5/6), or the stratified epithelial marker ΔNp63, but did express the mesenchymal markers vimentin and N-cadherin., Conclusion: Studies of epithelial carcinogenesis will benefit from culture systems which allow manipulation of the stromal and epithelial layers independently. We have developed an organotypic culture using primary oesophageal squamous cells and fibroblasts in which a stratified epithelium with a proliferative basal layer that stains strongly for ΔNp63 develops. This model will be suitable for the study of the molecular events in the development of Barrett's oesophagus. The most commonly used normal oesophageal squamous cell line, HET-1A, does not have the characteristics of normal oesophageal squamous cells and should not be used in models of the normal oesophageal epithelium. Until more representative cell lines are available, future studies in oesophageal cancer will be reliant on the availability and manipulation of primary tissue.
- Published
- 2010
- Full Text
- View/download PDF
18. Congenital heart block associated with Sjögren syndrome: case report.
- Author
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Moutasim KA, Shirlaw PJ, Escudier MP, and Poate TW
- Abstract
Background: Congenital heart block is a rare complication of pregnancy associated with Sjögren Syndrome that may result in the death of the foetus or infant, or the need for pacing in the newborn or at a later stage., Case Report: The case is presented of a 64-year-old patient with primary Sjögren Syndrome and a history of having given birth to two sons with congenital heart block, both of whom required pacing several years later., Conclusion: The literature relating to this association is discussed including the suggested mechanism, long-term outcome of mothers of children with congenital heart block and preventive treatment strategies.
- Published
- 2009
- Full Text
- View/download PDF
19. A case of vulvovaginal gingival lichen planus in association with Good's syndrome.
- Author
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Moutasim KA, Poate TW, Setterfield JF, and Challacombe SJ
- Subjects
- Female, Humans, Immunologic Deficiency Syndromes complications, Lichen Planus, Oral complications, Middle Aged, Syndrome, Agammaglobulinemia complications, Lichen Planus complications, Thymoma complications, Vaginal Diseases complications, Vulvar Diseases complications
- Abstract
Vulvovaginal gingival lichen planus (VVG LP) is a distinct variant of LP frequently associated with mucocutaneous scarring and vaginal stricture formation. Good's Syndrome (thymoma with hypogammaglobulinemia) is a rare cause of immunodeficiency in adults. The clinical features, investigation findings, and challenges in the management of a patient presenting with VVG LP and Good's Syndrome are discussed.
- Published
- 2008
- Full Text
- View/download PDF
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