c-KIT receptor expression is strictly associated with the biological behaviour of thyroid nodules

<p>Abstract</p> <p>Background</p> <p>A large amount of information has been collected on the molecular tumorigenesis of thyroid cancer. A low expression of c-KIT gene has been reported during the transformation of normal thyroid epithelium to papillary carcinoma suggesting a possible role of the gene in the differentiation of thyroid tissue rather than in the proliferation. The initial presentation of thyroid carcinoma is through a nodule and the best way nowadays to evaluate it is by fine-needle aspiration (FNA). However many thyroid FNAs are not definitively benign or malignant, yielding an indeterminate or suspicious diagnosis which ranges from 10 to 25% of FNAs. BRAF mutational analysis is commonly used to assess the malignancy of thyroid nodules but unfortunately it still leaves indeterminate diagnoses. The development of molecular initial diagnostic tests for evaluating a thyroid nodule is needed in order to define optimal surgical approach for patients with uncertain diagnosis pre- and intra-operatively.</p> <p>Methods</p> <p>In this study we extracted RNA from 82 FNA smears, 46 malignant and 36 benign at the histology, in order to evaluate by quantitative Real Time PCR the expression levels of c-KIT gene.</p> <p>Results</p> <p>We have found a highly preferential decrease rather than increase in transcript of c-KIT in malignant thyroid lesions compared to the benign ones. To explore the diagnostic utility of c-KIT expression in thyroid nodules, its expression values were divided in four arbitrarily defined classes, with class I characterized by the complete silencing of the gene. Class I and IV represented the two most informative groups, with 100% of the samples found malignant or benign respectively. The molecular analysis was proven by ROC (receiver operating characteristic) analysis to be highly specific and sensitive improving the cytological diagnostic accuracy of 15%.</p> <p>Conclusion</p> <p>We propose the use of BRAF test (after uncertain cytological diagnosis) to assess the malignancy of thyroid nodules at first, then the use of the c-KIT expression to ultimately assess the diagnosis of the nodules that otherwise would remain suspicious. The c-KIT expression-based classification is highly accurate and may provide a tool to overcome the difficulties in today's preoperative diagnosis of thyroid suspicious malignancies.</p

Cancer astrocytes have a more conserved molecular status in long recurrence free survival (RFS) IDH1 wild-type glioblastoma patients: New emerging cancer players

Glioblastoma is a devastating disease that despite all the information gathered so far, its optimal management remains elusive due to the absence of validated targets from clinical studies. A better clarification of the molecular mechanisms is needed. In this study, having access to IDH1 wild-type glioblastoma of patients with exceptionally long recurrence free survival (RFS), we decided to compare their mutational and gene expression profile to groups of IDH1 wild-type glioblastoma of patients with shorter RFS, by using NGS technology. The exome analysis revealed that Long-RFS tumors have a lower mutational rate compared to the other groups. A total of 158 genes were found differentially expressed among the groups, 112 of which distinguished the two RFS extreme groups. Overall, the exome data suggests that shorter RFS tumors could be, chronologically, in a more advanced state in the muli-step tumor process of sequential accumulation of mutations. New players in this kind of cancer emerge from the analysis, confirmed at the RNA/DNA level, identifying, therefore, possible oncodrivers or tumor suppressor genes

Energy Resolution Performance of the CMS Electromagnetic Calorimeter

The energy resolution performance of the CMS lead tungstate crystal electromagnetic calorimeter is presented. Measurements were made with an electron beam using a fully equipped supermodule of the calorimeter barrel. Results are given both for electrons incident on the centre of crystals and for electrons distributed uniformly over the calorimeter surface. The electron energy is reconstructed in matrices of 3 times 3 or 5 times 5 crystals centred on the crystal containing the maximum energy. Corrections for variations in the shower containment are applied in the case of uniform incidence. The resolution measured is consistent with the design goals

Analysis of shared common genetic risk between amyotrophic lateral sclerosis and epilepsy

Because hyper-excitability has been shown to be a shared pathophysiological mechanism, we used the latest and largest genome-wide studies in amyotrophic lateral sclerosis (n = 36,052) and epilepsy (n = 38,349) to determine genetic overlap between these conditions. First, we showed no significant genetic correlation, also when binned on minor allele frequency. Second, we confirmed the absence of polygenic overlap using genomic risk score analysis. Finally, we did not identify pleiotropic variants in meta-analyses of the 2 diseases. Our findings indicate that amyotrophic lateral sclerosis and epilepsy do not share common genetic risk, showing that hyper-excitability in both disorders has distinct origins

ANALYTICAL AND CLINICAL VALIDATION OF BIOMARKERS FOR NON INVASIVE EARLY DIAGNOSIS OF BLADDER CANCER

Bladder cancer (BC) is the seventh most common cancer worldwide and its incidence has increased in the last 10 years. It is predominantly a disease of older males, with a median age at presentation of 60-65 years. There are geographical variations in incidence, in Europe the highest incidence rates for men are in northern Italy, Spain and Geneva, Switzerland. The etiology is well defined, with the most common association being cigarette smoking and other factors including exposure to dyes and some industrial solvents, exhaust fumes. Nearly 80% of new cases originate from diffuse flat hyperplasia also referred to as low-grade intraurothelial neoplasia. They are typically of low histologic grade, growing as superficial papillary protrusions, and have high propensity for recurrence but they practically never invade the bladder wall or metastasize. The remaining 20% of tumors are non-papillary and develop from severe displasia or carcinoma in situ (CIS), also termed high-grade intraurothelial neoplasia. About 10-15% of low grade superficial papillary tumors progress to invade the bladder wall and may metastasize [1]. Both types of cancer are significant problem for public health: papillary tumors are not usually life threatening but because of their high recurrence rate, they contribute to bladder cancer’s rank as most expensive in terms of clinical management. Although about half of muscle-invasive tumors do initially respond cisplatin-based chemotherapy, the development of drug resistance is the major problem, and the disease progression in resistant tumors is rapid and uniformly fatal [2]. Many factors, such as chromosomal anomalies, genetic polymorphisms, genetic and epigenetic alterations, contribute to tumorigenesis and progression of bladder cancer. Several authors have reported that the detection of single genetic abnormalities can improve diagnosis and surveillance of the disease Different genes are well known to be involved independently in the development of BC, leading to loss of cell proliferation control (genes in FGFR3-p53 pathway), apoptosis resistance (i.e. Survivn) and promoting Epithelial Mesenchimal Transition (EMT) (i.e.: CK19, CK20, E cadherin and CD44). MicroRNAs (miRNAs) are part of a class of small ribonucleic acid (RNAs). They are important regulatory molecules, involved in several cell processes, such as developmental timing, stem cell division and apoptosis. Dysregulated miRNAs have been identified in several human malignancies, including bladder cancer tissue samples, and may confer a “tumour signature” that can be exploited for diagnostic purposes. To determine whether there is a correlation between the genetic profile and the histo-pathological feature of BC in an Italian population, we genetically characterized 66 Italian patients affected by BC, analyzing both TP53 and FGFR3 mutational status, and CK19-20, E cadherin, Survivin and CD44 gene expression levels. We report also a prospective pilot study investigating the diagnostic ability of a two miRNAs in voided urine samples collected from patients with bladder cancer just prior to bladder tumour resection Survivin was significantly over-expressed (p=0.019) in invasive tumors related to CK20 down-regulation (p=0.025). Using multivariate analysis we observed a significant positive relationship between CK19 and CK20 (p=0.0006), a significant association between E cadherin and CK20 in both High Grade (HG) and Low Grade (LG) tumors (p=0.0064 and p=0.0000, respectively) and a strong positive association between Survivin and CD44 (p= 0.0000) only in HG tumors. TP53 and FGFR3 mutations showed a heterogenic distribution. Gene expression data obtained by RT-PCR on CD44, E-cadherin and Survivin in urine sediments don’t show any significant value in discriminating BC patients from healthy donor group. On the contrary, the two miRNAs of interest were significantly up regulated in urine of BC patients than in healthy controls. Moreover, different statistical methods, like Discriminant Analysis (StatGraphics software) and Artificial Neural Network (ANN), showed a really strong predictive value (around 89% and 83% respectively) using miRNA expression value together in discriminating BC patents from healthy group. Taken together, our findings on TP53 and FGFR3 mutational status support the well-known genetic heterogeneity of BCs. However our results put in evidence the role of CD44, CK20, CK19 and Survivin on invasiveness ability and apoptosis resistance in HG tumors. The results on miRNA analysis provide rationale for further studies on validation of candidate miRNAs in voided urine and may potentially lead to the development of a non-invasive and sensitive test for bladder cancer diagnosis and recurrence surveillance

A molecular computational model improves the preoperative diagnosis of thyroid nodules

<p>Abstract</p> <p>Background</p> <p>Thyroid nodules with indeterminate cytological features on fine needle aspiration (FNA) cytology have a 20% risk of thyroid cancer. The aim of the current study was to determine the diagnostic utility of an 8-gene assay to distinguish benign from malignant thyroid neoplasm.</p> <p>Methods</p> <p>The mRNA expression level of 9 genes (KIT, SYNGR2, C21orf4, Hs.296031, DDI2, CDH1, LSM7, TC1, NATH) was analysed by quantitative PCR (q-PCR) in 93 FNA cytological samples. To evaluate the diagnostic utility of all the genes analysed, we assessed the area under the curve (AUC) for each gene individually and in combination. BRAF exon 15 status was determined by pyrosequencing. An 8-gene computational model (Neural Network Bayesian Classifier) was built and a multiple-variable analysis was then performed to assess the correlation between the markers.</p> <p>Results</p> <p>The AUC for each significant marker ranged between 0.625 and 0.900, thus all the significant markers, alone and in combination, can be used to distinguish between malignant and benign FNA samples. The classifier made up of KIT, CDH1, LSM7, C21orf4, DDI2, TC1, Hs.296031 and BRAF had a predictive power of 88.8%. It proved to be useful for risk stratification of the most critical cytological group of the indeterminate lesions for which there is the greatest need of accurate diagnostic markers.</p> <p>Conclusion</p> <p>The genetic classification obtained with this model is highly accurate at differentiating malignant from benign thyroid lesions and might be a useful adjunct in the preoperative management of patients with thyroid nodules.</p

Loss of c-KIT expression in thyroid cancer cells.

Papillary thyroid carcinoma is the most frequent histologic type of thyroid tumor. Few studies investigated the role of c-KIT expression in thyroid tumors, suggesting a role for this receptor and its ligand in differentiation and growth control of thyroid epithelium and a receptor loss following malignant transformation. We investigated and correlated c-KIT expression levels and two known markers of thyrocytes differentiation, PAX8 and TTF-1, in malignant and benign cytological thyroid samples. Moreover, we performed functional studies on human papillary thyroid carcinoma cell line to associated c-KIT expression to thyrocytes differentiation and tumor proliferation. c-KIT and PAX8 expression resulted higher in benign samples compared to the malignant ones, and the expression levels of these two genes were significantly correlated to each other. We also observed that c-KIT overexpression led to an increase of PAX8 expression level together with a decrease of proliferation. Furthermore, c-KIT overexpressing cells showed a regression of typical morphological features of malignancy. Taken together these results suggest that c-KIT could be involved in the differentiation of thyroid cells and in tumor progression

Molecular characterization of low grade and high grade bladder cancer

Background Bladder cancer (BC) is the 9 th most common cancer diagnosis worldwide. Low grade (LG) represents 70% of all BCs, characterized by recurrence and rare ability (10–15%) to progress to high grade (HG) and invade. The remaining 30% is high grade (HG), fast invasive BC, which is resistant to therapy. Identifying biomarkers for predicting those tumors able to progress is a key goal for patient outcome improvement. This study focuses on the most promising prognostic markers. Materials and methods TP53 and FGFR3 mutational status, Survivin, CK19, CK20, E-cadherin and CD44 gene expression analysis were performed on 66 BCs. Results Survivin was found associated to tumor grade (p<0.05). Moreover, Survivin correlated with CD44 in TP53 wild type (p = 0.0242) and FGFR3 wild type (p = 0.0036) tumors. In particular the Survivin-CD44 correlation was associated to HG FGFR3 wild type BCs (p = 0.0045). Unsupervised hierarchical clustering based on gene expression data identified four distinct molecular groups reflecting the patient histology (p = 0.038). Conclusion We suggest Survivin, both as a biomarker associated to G3 BCs but negatively related to TP53 mutational status, and as a potential novel therapeutic target

Loss of c-KIT expression in thyroid cancer cells

<div><p>Papillary thyroid carcinoma is the most frequent histologic type of thyroid tumor. Few studies investigated the role of c-KIT expression in thyroid tumors, suggesting a role for this receptor and its ligand in differentiation and growth control of thyroid epithelium and a receptor loss following malignant transformation. We investigated and correlated c-KIT expression levels and two known markers of thyrocytes differentiation, PAX8 and TTF-1, in malignant and benign cytological thyroid samples. Moreover, we performed functional studies on human papillary thyroid carcinoma cell line to associated c-KIT expression to thyrocytes differentiation and tumor proliferation. c-KIT and PAX8 expression resulted higher in benign samples compared to the malignant ones, and the expression levels of these two genes were significantly correlated to each other. We also observed that c-KIT overexpression led to an increase of PAX8 expression level together with a decrease of proliferation. Furthermore, c-KIT overexpressing cells showed a regression of typical morphological features of malignancy. Taken together these results suggest that c-KIT could be involved in the differentiation of thyroid cells and in tumor progression.</p></div

Effect of c-KIT overexpression on cell proliferation.

<p>WST-1 based proliferation assays were performed for control, empty vector transfected cells and c-KIT overexpressing cells.</p