Katherine Ashton

Lancashire Teaching Hospitals NHS Foundation Trust, Chorley, England, United Kingdom

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Publications (27)95.3 Total impact

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    Full-text · Dataset · Nov 2015
  • T. Mudariki · R.W. Lea · K. Ashton · T. Dawson · C. Davis · J.A. Smith

    No preview · Article · Nov 2015 · Neuro-Oncology

  • No preview · Article · Nov 2015 · Neuro-Oncology
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    ABSTRACT: Molecular biomarkers are commonly used for the management of several types of malignant tumours in routine clinical practice. However, this is not the case for malignant gliomas. Cytokines and Angiogenesis factors are potential candidates due to their intrinsic role in tumourigenesis. Pre- and post-operative serum from 36 malignant glioma patients and 36 controls was analysed using the Bio-Plex Pro Angiogenesis and Cytokines Assay (Bio-Rad, USA). Amongst the molecules tested, the serum concentration of follistatin was significantly higher in patients than in controls. Moreover, the serum concentration of follistatin of the patients postoperatively was significantly reduced compared to that preoperatively. Factors such as age and gender did not affect the concentrations of follistatin measured in the serum of patients pre- and post-operatively as well as healthy controls. This is the first report of follistatin as potential biomarker for the detection of malignant gliomas.
    Full-text · Article · Sep 2015 · Neuroscience & Medicine
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    ABSTRACT: Malignant glioma is characterised by a rapid growth rate and high capacity for invasive infiltration to surrounding brain tissue; hence, diagnosis and treatment is difficult and patient survival is poor. Aptamers contribute a promising and unique technology for the in vitro imaging of live cells and tissues, with a potentially bright future in clinical diagnostics and therapeutics for malignant glioma. The binding selectivity, uptake capacity and binding target of two DNA aptamers, SA43 and SA44, were investigated in glioma cells and patient tissues. The binding assay showed that SA43 and SA44 bound with strong affinity (K d , 21.56 ± 4.60 nM and K d , 21.11 ± 3.30 nM respectively) to the target U87MG cells. Quantitative analysis by flow cytometry showed that the aptamers were able to actively internalise in U87MG and 1321N1 glioma cells compared to the non-cancerous and non-glioma cell types. Confocal microscopy confirmed staining in the cytoplasm, and co-localisation studies with endoplasmic reticulum, Golgi apparatus and lysosomal markers suggested internalisa-tion and compartmentalisation within the endomembrane system. Both aptamers selectively bound to Ku 70 and Ku 80 DNA repair proteins as determined by aptoprecipitation (AP) followed by mass spectrometry analysis and confirmation by Western blot. In addition, aptohistochemical (AHC) staining on paraffin embedded, formalin fixed patient tissues revealed that the binding selectivity was significantly higher for SA43 aptamer in glioma tissues (grade I, II, III and IV) compared to the non-cancerous tissues, whereas SA44 did not show selectivity towards glioma tissues. The results indicate that SA43 aptamer can differentiate between glioma and non-cancerous cells and tissues and therefore, shows promise for histological diagnosis of glioma.
    Full-text · Article · Aug 2015 · PLoS ONE
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    ABSTRACT: Tumour metastasis to the brain is a common and deadly development in certain cancers; 18-30 % of breast tumours metastasise to the brain. The contribution that gene silencing through epigenetic mechanisms plays in these metastatic tumours is not well understood. We have carried out a bioinformatic screen of genome-wide breast tumour methylation data available at The Cancer Genome Atlas (TCGA) and a broad literature review to identify candidate genes that may contribute to breast to brain metastasis (BBM). This analysis identified 82 candidates. We investigated the methylation status of these genes using Combined Bisulfite and Restriction Analysis (CoBRA) and identified 21 genes frequently methylated in BBM. We have identified three genes, GALNT9, CCDC8 and BNC1, that were frequently methylated (55, 73 and 71 %, respectively) and silenced in BBM and infrequently methylated in primary breast tumours. CCDC8 was commonly methylated in brain metastases and their associated primary tumours whereas GALNT9 and BNC1 were methylated and silenced only in brain metastases, but not in the associated primary breast tumours from individual patients. This suggests differing roles for these genes in the evolution of metastatic tumours; CCDC8 methylation occurs at an early stage of metastatic evolution whereas methylation of GANLT9 and BNC1 occurs at a later stage of tumour evolution. Knockdown of these genes by RNAi resulted in a significant increase in the migratory and invasive potential of breast cancer cell lines. These findings indicate that GALNT9 (an initiator of O-glycosylation), CCDC8 (a regulator of microtubule dynamics) and BNC1 (a transcription factor with a broad range of targets) may play a role in the progression of primary breast tumours to brain metastases. These genes may be useful as prognostic markers and their products may provide novel therapeutic targets.
    Full-text · Article · May 2015
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    ABSTRACT: Cyclic nucleotides (cAMP & cGMP) are critical intracellular second messengers involved in the transduction of a diverse array of stimuli and their catabolism is mediated by phosphodiesterases (PDEs). We previously detected focal genomic amplification of PDE1C in >90 glioblastoma multiforme (GBM) cells suggesting a potential as a novel therapeutic target in these cells. In this report, we show that genomic gain of PDE1C was associated with increased expression in low passage GBM-derived cell cultures. We demonstrate that PDE1C is essential in driving cell proliferation, migration and invasion in GBM cultures since silencing of this gene significantly mitigates these functions. We also define the mechanistic basis of this functional effect through whole genome expression analysis by identifying down-stream gene effectors of PDE1C which are involved in cell cycle and cell adhesion regulation. In addition, we also demonstrate that Vinpocetine, a general PDE1 inhibitor, can also attenuate proliferation with no effect on invasion/migration. Up-regulation of at least one of this gene set (IL8, CXCL2, FOSB, NFE2L3, SUB1, SORBS2, WNT5A, and MMP1) in TCGA GBM cohorts is associated with worse outcome and PDE1C silencing down-regulated their expression, thus also indicating potential to influence patient survival. Therefore we conclude that proliferation, migration, and invasion of GBM cells could also be regulated downstream of PDE1C. © 2015 Wiley Periodicals, Inc.
    No preview · Article · Jan 2015 · Molecular Carcinogenesis
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    ABSTRACT: Tumor metastasis to the brain is a common and deadly development in certain cancers; 18-30% of breasts tumors metastasize to the brain. The contribution that gene silencing through epigenetic mechanisms plays in metastasis to the brain is not well understood. To identify epigenetic drivers of brain metastasis we have combined a candidate gene screen and Genome-wide methylation analysis of metastatic brain tumors that originated from primary breast tumors. From a screen of 85 candidates and an independent comparison of genome-wide methylation profiles in brain metastases we have identified genes that are frequently methylated in breast-to-brain metastases and infrequently methylated in primary breast tumors. We have identified gene methylation that either occurs early in tumour evolution (methylation is present in primary tumors that metastasize) or late in tumour evolution (methylation is only present in the brain metastasis). Many of the genes identified are involved in transcriptional control, either by DNA binding of through mRNA regulation (MicroRNAs etc.). We are carrying out in vitro assays to determine if epigenetic deregulation of these genes contributes to the metastatic phenotype: CCDC8 and GALNT9 are methylated in 87% and 53% of breast-to brain metastases respectively, knock-down of these genes by RNAi resulted in a significant migratory (p = 0.0033, p = 0.0253) advantage of breast cancer cell lines. BNC1 and L3MBTL1 are methylated in 73% and 67% of breast-to brain metastases respectively, knock-down by RNAi resulted in a significant migratory (p = 0.0302, p = 0.0061) and invasive (p = 0.0012, p = 0.0161) advantage to breast cancer cell lines. These early data suggest that this approach has the potential to identify genes deregulated in the process of tumour metastasis to the brain. Further analysis is required to determine if these genes may be used as therapeutic targets or prognostic markers.
    No preview · Conference Paper · Nov 2014
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    ABSTRACT: INTRODUCTION: The ability to diagnose brain tumours rapidly from serum would allow for rapid testing and decreased time to results providing a responsive diagnostic environment. ATR-FTIR (Attenuated Total Reflection - Fourier Transform Infrared Spectroscopy) generates a serum sample - fingerprint - since biomolecules exhibit different responses to different wavelengths of light. We have previously used ATR-FTIR to discriminate high grade brain cancer, low-grade brain cancer and normal serum to sensitivities and specificities on average of 93.75 and 96.53 % respectively. This paper reports the analysis of 429 patients. METHOD: 1 µl of serum was analysed in triplicate from 429 patients via ATR-FTIR. This dataset was analysed using pattern recognition algorithms for the following models 1) Cancer (n = 314) vs Non-cancer (n = 122), 2) Metastatic Brain Cancer (n = 192) vs Brain Cancer (n = 124) vs Non-Cancer (n = 122), 3) Organ of tumour origin, 4)High grade glioma (n = 52) vs low grade glioma (n = 24) vs meningioma (n = 47) 5) Subtype of Brain Cancer. RESULTS: ATR-FTIR spectroscopy was able to discriminate different groupings with high accuracy. The predicted range for each diagnostic layer was between 78.26 - 100.00 % sensitivity and specificity respectively. Interrogation of the loadings plots showed differences in Amide and lipid content of the serum. CONCLUSION: A robust reproducible method has been developed for the diagnosis of small volumes of serum. In addition multiple layers of diagnostic information can be provided from the same spectral dataset. ATR-FTIR has shown to be an excellent clinical tool for rapid diagnosis enabling a dynamic and responsive clinical environment. Future work will investigate spectroscopic changes during treatment.
    Full-text · Article · Oct 2014 · Neuro-Oncology
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    ABSTRACT: INTRODUCTION: Complete tumour removal during surgery is one of the primary factors for recurrence-free survival. The current process is unable to accurately identify tumour margins and data feedback takes too long for responsive action during surgery. Our recent study has shown the ability of Raman spectroscopy to distinguish between metastatic brain, GBM and normal brain tissue to sensitivities and specificities ranging from 85.71% to 100%. Pushing this result towards the clinic needs an assessment of the data feedback. In a surgery situation, audio feedback of tumour margins would provide a real time response allowing the surgeons visual focus to remain on the patient.Auditory perception of complex, structured information could have several advantages when compared to visual images including the capability of the human ear to detect patterns, recognise timbres and follow different strands at the same time. METHOD: Our method consists of two phases, to compute the relevant variables, we apply feature extraction to the data, extracting band-wise spectral shape descriptors such as centroid, skew and kurtosis. These are then used to modulate parameters of a synthesizer. RESULTS: The method was applied to a dataset of 311 samples, 136 from an uncontrolled and 176 from a controlled group. Based on subjective listening tests taken from around 20 participants, we were able to get manual classification accuracies of up to 83%, using a multi-stimulus technique. CONCLUSION: Raman spectroscopy is able to accurately distinguish between cancerous and non-cancerous tissue. The development of novel data extraction tools has enabled audio feedback of these molecular differences in real time allowing the surgeons visual focus to remain on the patient.
    Full-text · Article · Oct 2014 · Neuro-Oncology

  • No preview · Article · Jul 2014 · Neuro-Oncology
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    ABSTRACT: Gliomas are the most frequent primary brain tumours in adults with over 9,000 people diagnosed each year in the UK. A rapid, reagent-free and cost-effective diagnostic regime using serum spectroscopy would allow for rapid diagnostic results and for swift treatment planning and monitoring within the clinical environment. We report the use of ATR-FTIR spectral data combined with a RBF-SVM for the diagnosis of gliomas (high-grade and low-grade) from non-cancer with sensitivities and specificities on average of 93.75 and 96.53% respectively. The proposed diagnostic regime has the ability to reduce mortality and morbidity rates. (© 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim).
    Full-text · Article · Apr 2014 · Journal of Biophotonics
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    Full-text · Conference Paper · Mar 2014
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    ABSTRACT: It is estimated that approximately 13 000 people in the UK are diagnosed with brain cancer every year; of which 60% are metastatic. Current methods of diagnosis can be subjective, invasive and have long diagnostic windows. Raman spectroscopy provides a non-destructive, non-invasive, rapid and economical method for diagnosing diseases. The aim of this study was to investigate the use of Raman and immersion Raman spectroscopy for diagnosing metastatic brain cancer and identifying primary sites of origin using brain tissue. Through spectral examination, the Raman peaks at 721 cm(-1) and 782 cm(-1) were identified as being the most distinct for discriminating between the glioblastoma multiforme (GBM), metastatic and normal brain tissue spectra. A ratio score plot of these peaks calculated the classification sensitivities and specificities as 100% and 94.44% for GBM, 96.55% and 100% for metastatic brain, and 85.71% and 100% for normal brain tissue. Principal Component-Linear Discriminant Analysis (PC-LDA) also showed discrimination between normal, GBM and metastatic brain tissue spectra. We also present, for the first time, the use of Raman spectroscopy to investigate primary site of origin for metastatic brain cancer and any biochemical differences between different primary and metastatic cancer using linked samples. This study revealed interesting spectral differences in the amide regions showing changes in the biochemistry of the metastatic brain cancer from the primary cancer.
    No preview · Article · Jan 2014 · Analytical methods
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    ABSTRACT: Raman spectroscopy is a non-destructive, non-invasive, rapid and economical technique which has the potential to be an excellent method for the diagnosis of cancer and understanding disease progression through retrospective studies of archived tissue samples. Historically, biobanks are generally comprised of formalin fixed paraffin preserved tissue and as a result these specimens are often used in spectroscopic research. Tissue in this state has to be dewaxed prior to Raman analysis to reduce paraffin contributions in the spectra. However, although the procedures are derived from histopathological clinical practice, the efficacy of the dewaxing procedures that are currently employed is questionable. Ineffective removal of paraffin results in corruption of the spectra and previous experiments have shown that the efficacy can depend on the dewaxing medium and processing time. The aim of this study was to investigate the influence of commonly used spectroscopic substrates (CaF2, Spectrosil quartz and low-E slides) and the influence of different histological tissue types (normal, cancerous and metastatic) on tissue preparation and to assess their use for spectral histopathology. Results show that CaF2 followed by Spectrosil contribute the least to the spectral background. However, both substrates retain paraffin after dewaxing. Low-E substrates, which exhibit the most intense spectral background, do not retain wax and resulting spectra are not affected by paraffin peaks. We also show a disparity in paraffin retention depending upon the histological identity of the tissue with abnormal tissue retaining more paraffin than normal.
    No preview · Article · Dec 2013 · The Analyst
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    ABSTRACT: The ability to diagnose brain cancer rapidly from serum samples is of great interest; such a diagnosis would allow for rapid testing and time to results providing a responsive diagnostic environment, ability to monitor treatment efficacy, early detection of recurrent tumours and screening techniques. Current methods rely upon subjective, time-consuming tests such as histological grading and are particularly invasive with the diagnostic test requiring hospitalisation of 2–3 days. A rapid diagnostic method based upon serum samples would allow for a relatively non-invasive test and open up the possibility of screening for brain cancer. We report for the first time the use of a Bioplex immunoassay to provide cytokine and angiogenesis factor levels that differ between serum from glioma and non-cancer patients specifically angiopoietin, follistatin, HGF, IL-8, leptin, PDGF-BB and PECAM-1 providing sensitivities and specificities as high as 88 % and 81 %, respectively. We also report, for the first time, the use of serum ATR-FTIR combined with a RBF SVM for the diagnosis of gliomas from non-cancer patients with sensitivities and specificities as high as 87.5 % and 100 %, respectively. We describe the combination of these techniques in an orthogonal diagnostic regime, providing strength to the diagnosis through data combinations, in a rapid diagnostic test within 5 h from serum collection (10 min for ATR-FTIR and 4 h for the Bioplex Immunoassay). This regime has the ability to revolutionise the clinical environment by providing objective measures for diagnosis allowing for increased efficiency with corresponding decreases in mortality, morbidity and economic impact upon the health services. Figure ᅟ
    Full-text · Article · Jul 2013 · Analytical and Bioanalytical Chemistry
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    ABSTRACT: The most common initial treatment received by patients with a brain tumour is surgical removal of the growth. Precise histopathological diagnosis of brain tumours is to some extent subjective. Furthermore, currently available diagnostic imaging techniques to delineate the excision border during cytoreductive surgery lack the required spatial precision to aid surgeons. We set out to determine whether infrared (IR) and/or Raman spectroscopy combined with multivariate analysis could be applied to discriminate between normal brain tissue and different tumour types (meningioma, glioma and brain metastasis) based on the unique spectral "fingerprints" of their biochemical composition. Formalin-fixed paraffin-embedded tissue blocks of normal brain and different brain tumours were de-waxed, mounted on low-E slides and desiccated before being analyzed using attenuated total reflection Fourier-transform IR (ATR-FTIR) and Raman spectroscopy. ATR-FTIR spectroscopy showed a clear segregation between normal and different tumour subtypes. Discrimination of tumour classes was also apparent with Raman spectroscopy. Further analysis of spectral data revealed changes in brain biochemical structure associated with different tumours. Decreased tentatively-assigned lipid-to-protein ratio was associated with increased tumour progression. Alteration in cholesterol esters-to-phenylalanine ratio was evident in grade IV glioma and metastatic tumours. The current study indicates that IR and/or Raman spectroscopy have the potential to provide a novel diagnostic approach in the accurate diagnosis of brain tumours and have potential for application in intra-operative diagnosis.
    Full-text · Article · Jan 2013 · Analytical methods
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    ABSTRACT: The most common initial treatment received by patients with a brain tumour is surgical removal of the growth. Precise histopathological diagnosis of brain tumours is to some extent subjective. Furthermore, currently available diagnostic imaging techniques to delineate the excision border during cytoreductive surgery lack the required spatial precision to aid surgeons. We set out to determine whether infrared (IR) and/or Raman spectroscopy combined with multivariate analysis could be applied to discriminate between normal brain tissue and different tumour types (meningioma, glioma and brain metastasis) based on the unique spectral "fingerprints" of their biochemical composition. Formalin-fixed paraffin-embedded tissue blocks of normal brain and different brain tumours were de-waxed, mounted on low-E slides and desiccated before being analyzed using attenuated total reflection Fourier-transform IR (ATR-FTIR) and Raman spectroscopy. ATR-FTIR spectroscopy showed a clear segregation between normal and different tumour subtypes. Discrimination of tumour classes was also apparent with Raman spectroscopy. Further analysis of spectral data revealed changes in brain biochemical structure associated with different tumours. Decreased tentatively-assigned lipid-to-protein ratio was associated with increased tumour progression. Alteration in cholesterol esters-to-phenylalanine ratio was evident in grade IV glioma and metastatic tumours. The current study indicates that IR and/or Raman spectroscopy have the potential to provide a novel diagnostic approach in the accurate diagnosis of brain tumours and have potential for application in intra-operative diagnosis.
    No preview · Article · Sep 2012 · Analytical methods
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    ABSTRACT: Glioblastoma multiforme is a malignant primary brain tumour with very limited treatment options. Any addition to existing treatment options which can improve prognosis and life expectancy is useful. In our study, we look at the usefulness of anti-progestogen mifepristone in causing growth suppression of glioma cell lines in the laboratory. We cultured five cell lines in the lab and exposed them to mifepristone in different doses for a total of 96 h. Five different doses of mifepristone were used. Progesterone and dexamethasone were also used as growth stimulants. Immunostaining was used to identify progesterone receptors (PRs) in the cell lines. U257/7 and IN1265 showed statistically significant growth suppression (36% and 11%, P = 0.001 and 0.03 respectively), maximal at 96 h. Growth suppression in U257/7 showed a dose response progression except with the lowest dose which was not explicable. The response of IN1265 was seen only with the highest dose of mifepristone. There was no significant growth stimulation with either dexamethasone or progesterone. None of the cell lines showed any significant positivity for PRs. We were able to produce enough growth suppression of glioma cell lines using mifepristone. This is in keeping with some of the published results in literature. This raises the possibility of using mifepristone in treating GBMs which have very limited treatment options. This, however, needs further work probably on primary glioma cultures first followed by in vivo studies before it can be used in patients.
    No preview · Article · Nov 2011 · British Journal of Neurosurgery
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    ABSTRACT: Endometriosis is the growth of endometrial tissue outside of the uterine cavity. Its aetiology remains obscure, and it is difficult to diagnose ranging from asymptomatic to debilitating disease. Mid-infrared (IR) spectroscopy has become recognised as a potential clinical diagnostic tool. Biomolecules absorb mid-IR (4000 cm(-1) to 400 cm(-1)) and from this, a biochemical-cell fingerprint in the form of an absorbance spectrum can be derived. We set out to determine if IR spectroscopy could be used to identify underlying biochemical differences between endometrial tissues growing outside of the uterus (ectopic) from endometrial tissue of the uterus (eutopic). For comparative purposes, endometrial tissues from endometriosis-free women were also obtained (benign eutopic). Attenuated total reflection Fourier-transform IR (ATR-FTIR) spectroscopy or transmission FTIR microspectroscopy was employed for spectral acquisition. Principal component analysis (PCA)-linear discriminant analysis (LDA) was used for chemometric analysis. A clear segregation was exhibited between the three categories independent of inter-individual confounding differences. Importantly, there was a marked difference between eutopic endometrial tissue from patients with or without endometriosis. This indicates that IR spectroscopy coupled with multivariate analysis (e.g., PCA-LDA) may provide a non-invasive diagnostic tool for endometriosis. By analysing the underlying biochemistry of these endometrial tissues, this approach may facilitate a better understanding of this pathology.
    No preview · Article · Mar 2011 · The Analyst

Publication Stats

189 Citations
95.30 Total Impact Points

Institutions

  • 2004-2015
    • Lancashire Teaching Hospitals NHS Foundation Trust
      Chorley, England, United Kingdom
  • 2014
    • University of Wolverhampton
      • School of Applied Sciences
      Wolverhampton, England, United Kingdom
    • University of Central Lancashire
      Preston, England, United Kingdom