Histopat Laboratoris

Barcelona, Spain

Histopat Laboratoris

Barcelona, Spain
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Garcia-Pelaez B.,Biopatologia Molecular SL | Trias I.,Histopat Laboratoris | Trias I.,Hospital Plato Fundacio Privada | Roman R.,Biopatologia Molecular SL | And 6 more authors.
Actas Urologicas Espanolas | Year: 2013

Objective: To assess the value of the study of chromosomal alterations by fluorescent in situ hybridization in a series of patients diagnosed of urothelial carcinoma and a minimum follow up of twenty four months, as well as evaluate its putative predictive potential. Material and methods: The overall series includes 338 samples from 98 patients with 84 episodes of urothelial carcinoma. A subgroup of 24 patients who had at least one recurrence during the follow up was used to evaluate the predictive potential of the test. Three categories were considered (positive coherent episode, negative coherent episode, and incoherent episode) depending on the relationship between the fluorescent in situ hybridization result in the concomitant study of the new episode and those of the preceding samples. Results: Fluorescent in situ hybridization showed higher sensitivity regardless of grade, negative predictive value and accuracy, while specificity and positive predictive value were superior with conventional cytology. In the recurrence, series 19/29 episodes were coherent, 11/19 were positive coherent with urothelial carcinoma all high grade and 8/19 negative coherent, most low grade. Conclusions: Fluorescent in situ hybridization test shows good sensitivity during a follow up of twenty four months and is able to predict recurrence, especially in cases of high grade. Our data demonstrate the existence of urothelial carcinomas without detectable chromosomal abnormalities by currently available methodology. The results support a multidisciplinary follow up combining fluorescent in situ hybridization, cytology and cystoscopy. © 2012 AEU. Published by Elsevier España, S.L. All rights reserved.

Verdu M.,BIOPAT | Verdu M.,Histopat Laboratoris | Trias I.,BIOPAT | Trias I.,Histopat Laboratoris | And 11 more authors.
Virchows Archiv | Year: 2013

ERG gene rearrangement has been identified as a highly specific alteration that is present in 40-50 % of prostate carcinomas. The standardization of an immunohistochemical assay with a novel anti-ERG antibody recently described would have significant diagnostic value. The aims of this study were to identify the incidence of this rearrangement in a Spanish population and to test the specificity of immunohistochemical ERG evaluation for prostate carcinomas. Three prostate tissue microarrays were constructed using radical prostatectomy specimens and related to grade, local invasion, and regional invasion. In addition to samples from malignant cases (160), specimens of prostatic hyperplasia (26) and high-grade prostatic intraepithelial neoplasia (10) were included. Tissue microarrays of 270 samples from most common malignant tumors (breast, colon, lung, and bladder) were also tested. All were analyzed by immunohistochemistry. Seventy-five out of 154 evaluable cases (49 %) of prostate carcinoma showed ERG expression; 52/75 showed strong staining. No ERG expression was observed in any of the high-grade prostatic intraepithelial neoplasia. ERG expression was independent of Gleason score (p = 0.160), extent of invasion (p = 0.517), and regional lymph node involvement (p = 0.816). No ERG expression was found in any other type of tumor, with the exception of one bladder cancer sample that showed focal and weak expression. The frequency of ERG detected in our study correlated with the results published for other Caucasian populations. Strong ERG protein expression was exclusively detected in prostate carcinomas, corroborating the specificity of ERG rearrangements for these tumors. Thus, detecting ERG using immunohistochemistry may be useful in routine practice in pathology departments. © 2013 Springer-Verlag Berlin Heidelberg.

Verdu M.,Histopat Laboratoris | Trias I.,Histopat Laboratoris | Trias I.,Hospital Of Barcelona | Roman R.,BIOPAT | And 9 more authors.
Applied Immunohistochemistry and Molecular Morphology | Year: 2015

The coexpression of HER2 and EGFR L858R in a solitary nodule removed from the lung, whose mutation was not confirmed by molecular techniques, made us think about the possible existence of a cross-reaction between HER2 and the EGFR L858R-specific antibody. Our study was designed to further analyze the existence of this cross-reaction and stress the need to exclude a metastatic breast cancer when dealing with EGFR L858R-positive cases. The series consists of 42 primary breast carcinomas, 22 HER2 positive for overexpression and amplification, and 20 negative for both. EGFR mutations were studied by immunohistochemistry and confirmed using real-time PCR when positive. Immunohistochemistry assay with EGFR L858R was positive in 19 (86%) of the HER2-positive breast carcinomas and negative in all HER2-negative carcinomas. The EGFR L858R antibody gives false-positive results in most of the breast carcinomas with HER2 overexpression/amplification. As a consequence, it is essential to confirm any EGFR L858R-positive cases by molecular methods or at least discard the presence of HER2 overexpression/amplification before rendering a diagnosis. It is also important to consider that HER2 has been described in other carcinomas such as urothelial, gastric or ovarian, as well as lung, although infrequently. © Copyright © 2014 Wolters Kluwer Health, Inc. All rights reserved.

Garcia-Pelaez B.,BIOPAT | Verdu M.,BIOPAT | Verdu M.,Histopat Laboratoris | Roman R.,BIOPAT | And 6 more authors.
Revista Espanola de Patologia | Year: 2015

Sentinel lymph node assessment is an essential part of the staging procedure in breast cancer. Increasingly, One-Step Nucleic Acid Amplification (OSNA), an intraoperative molecular assay that measures the amount of cytokeratin 19 (CK19) mRNA present in the whole of the sentinel lymph node, is being used in the detection of sentinel lymph node metastasis, due to its superior sensitivity and equal specificity compared with conventional histological examination. CK19 positive immunostaining in a previous biopsy is recommended to avoid false negative results with OSNA analysis.The aim of our study was to assess the degree of CK19 positivity in series of breast carcinomas with particular emphasis on special histological subtypes. The total of 337 breast carcinomas studied were distributed in three series as follows: a retrospective unselected series studied on tissue microarray, a series of whole sections enriched for special histological subtypes and triple negative carcinomas, and a prospective series of biopsies studied in whole sections, prior to a routine OSNA assay.The results of our three series revealed that the great majority of breast carcinomas exhibits a diffuse CK19 positivity. Therefore it would be highly unlikely, especially in frequent variants, to have a false negative OSNA result due to lack of CK19 expression in the primary tumour. Heterogeneous expression was only observed in patients with infrequent variants (apocrine, adenoid cystic and medullary carcinomas) and/or TN profile. © 2014 SEAP y SEC.

Verdu M.,Histopat Laboratoris | Trias I.,Histopat Laboratoris | Trias I.,Hospital Of Barcelona | Roman R.,BIOPAT Biopatologia Molecular | And 7 more authors.
Revista Espanola de Patologia | Year: 2016

The study of epidermal growth factor receptor mutations has become essential for the treatment of lung cancer. The aim of this study was to find a correlation between morphological changes and EGFR mutational status using both immunohistochemistry and molecular techniques. We also analyzed the cross-reaction of the L858R mutation-specific monoclonal antibody in cases with HER2 amplification described previously in breast and gastric cancer.A series of 100 primary lung adenocarcinomas were examined. Exon 19 E746_A750del and exon 21 L858R mutations were studied using immunohistochemistry with two specific monoclonal antibodies. Gene mutational status was determined using real-time PCR or Sanger sequencing followed by real-time PCR when negative.EGFR mutations were detected in 22 cases (22%) by molecular techniques, being significantly more frequent in women, low grade carcinoma and lepidic subtype, (p-value <0.05 in all cases).In addition, in our series presence of tumoral necrosis correlated with absence of mutations.The anti-E746_A750del antibody achieved a 100% positive predictive value and a negative predictive value of 97.7% which could restrict the use of molecular techniques to the 7% of cases with an equivocal result. The antibody for L858R mutation showed inconsistent results compared to molecular techniques, giving false positive result in two adenocarcinomas with HER2 amplification. However, its negative predictive value was very high (98.9%).The use of real-time PCR identifies mutations not detected by the other two techniques.These new antibodies could be useful as a screening tool prior to EGFR molecular techniques. © 2016 Sociedad Española de Anatomía Patológica.

High microsatellite instability (MSI-H) allows the identification of a subset of colorectal carcinomas associated with good prognosis and a higher incidence of Lynch syndrome. The aim of this work was to assess the interobserver variability and optimize our MSI-H prediction model previously published based on phenotypic features. The validation series collected from five different hospitals included 265 primary colorectal carcinomas from the same number of patients. The eight clinicopathological parameters that integrate our original model were evaluated in the corresponding centers. Homogeneity assessment revealed significant differences between hospitals in the estimation of the growth pattern, presence of Crohn-like reaction, percentage of cribriform structures, and Ki-67 positivity. Despite this observation, our model was globally able to predict MSI-H with a negative predictive value of 97.0%. The optimization studies were carried out with 615 cases and resulted in a new prediction model RERtest8, which includes the presence of tumor infiltrating lymphocytes at the expense of the percentage of cribriform structures. This refined model achieves a negative predictive value of 97.9% that is maintained even when the immunohistochemical parameters are left out, RERtest6. The high negative predictive value achieved by our models allows the reduction of the cases to be tested for MSI to less than 10%. Furthermore, the easy evaluation of the parameters included in the model renders it a useful tool for the routine practice and can reinforce other published models and the current clinical protocols to detect the subset of colorectal cancer patients bearing hereditary nonpolyposis colorectal cancers risk and/or MSI-H phenotype. © Springer-Verlag 2010.

Verdu M.,Histopat Laboratoris | Trias I.,Histopat Laboratoris | Trias I.,Hospital Of Barcelona | Roman R.,BIOPAT | And 3 more authors.
Revista Espanola de Patologia | Year: 2015

Recently, the study of the EGFR mutations in lung cancer has become mandatory due to their usefulness in target therapy. Lately, the use of specific antibodies to study the most prevalent of these mutations has been introduced. The newly published discovery of the existence of a cross-reaction between the HER2 protein and the EGFR L858R-specific antibody in breast cancer led us to corroborate this cross-reactivity in other tumors; specifically, we have chosen gastric carcinomas where HER2 is routinely evaluated as a molecular therapy driver.Our series consists of 15 primary gastric carcinomas, 7 HER2 cases positive for overexpression/amplification and 8 negative cases for both techniques. EGFR L858R mutation was studied with immunohistochemistry and complemented with real time PCR when positive. Immunohistochemistry assay with EGFR L858R was positive in 5 of the HER2 positive carcinomas (71%), none of which was confirmed by PCR, and negative in all HER2 negative carcinomas. Conclusions: The EGFR L858R antibody gives false positive results in most gastric carcinomas with HER2 overexpression/amplification, confirming the results described previously in breast cancer. It is also important to bear in mind that HER2 has also been described in other carcinomas, including lung cancer. © 2014 SEAP y SEC.

Verdu M.,Biopatologia Molecular SL | Verdu M.,Histopat Laboratoris | Roman R.,Biopatologia Molecular SL | Calvo M.,University of Barcelona | And 8 more authors.
Modern Pathology | Year: 2011

Invasive micropapillary carcinoma is associated with frequent lymph node metastasis and adverse clinical outcome. Initially described as a variant of breast and ovarian carcinoma, it has subsequently been found in other organs, most recently the colon. Reports of colorectal micropapillary carcinoma to date are limited in number, and their molecular profile has not been established. The aims of the present study were to analyze their clinicopathological features and molecular profile, and compare them with those of conventional adenocarcinoma. Clinicopathological features of a cohort of 379 patients with primary colorectal cancer were retrospectively reviewed for the presence of the pattern characteristic of micropapillary carcinoma. We also assessed the expression of KRT7, KRT20, CEACAM5, MUC1 (EMA, clone E29), MUC1 (clone MA695), MLH1, MSH2, MSH6 and TP53 by immunohistochemistry. Genetic assessments of microsatellite instability, chromosomes 17p and 18q, and mutations in TP53, BRAF and KRAS were performed using DNA extracted from formalin-fixed, paraffin-embedded sections. In all, 60 of the reviewed cases (16%) had a micropapillary component that ranged from 5 to 95% of the tumor, characterized by a higher frequency of an infiltrative pattern, lymphovascular and perineural invasion, a higher depth of invasion and more positive lymph nodes than conventional adenocarcinoma. Immunohistochemistry for MUC1 (clone MA695) and MUC1 (EMA, clone E29) enhanced the characteristic inside-out staining pattern of the micropapillary carcinoma component, whereas the rest of the tumor showed luminal staining patterns. KRT7 expression was slightly increased in micropapillary carcinoma, but did not reach significance (17-3%, P0.1967). The molecular parameters showed a higher frequency of TP53 alterations and a low incidence of microsatellite instability and RER phenotype (loss of mismatch repair protein) in micropapillary carcinoma. With regard to the histological parameters, micropapillary carcinoma appears to be more aggressive than conventional colorectal adenocarcinoma. The molecular profile supports the hypothesis that micropapillary carcinoma carcinogenesis develops through the classical chromosomal instability pathway. © 2011 USCAP, Inc. All rights reserved.

Roman R.,Biopatologia Molecular SL | Rodon N.,Biopatologia Molecular SL | Verdu M.,Biopatologia Molecular SL | Verdu M.,Histopat Laboratoris | And 8 more authors.
Revista Espanola de Patologia | Year: 2015

Determining microsatellite instability status (MSI) is now highly recommended for all diagnosed colorectal carcinomas, both as a first approach to identify putative Lynch syndrome patients, and as a valuable prognostic indicator as it is associated with a poor response to 5-fluorouracil-based regimes. However, generally the implementation of MSI testing is still quite limited, suggesting the need for screening tools which would simplify MSI testing. This study presents the validation of two previously published prediction models, RERtest6 and RERtest8, based on clinicopathological features and without age restriction. The series includes 206 primary colorectal tumors from 199 patients, to which the models containing 6 or 8 parameters were applied before the assessment of MSI status, using the consensus NCI panel or the MSI Promega kit. High-level microsatellite instability (MSI-H) was detected in 21 cases (10.1%). Both models confirmed their reliability and were able to maintain negative predictive values close to 95%, reducing the number of cases to be tested by molecular methods to 10%. Furthermore, the nature of the parameters included in the models, mostly already part of a routine histopathological examination, makes them a useful tool that can be easily implemented into routine practice. © 2014 SEAP y SEC.

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