Publications by authors named "Esther Conde"

39 Publications

Screening for fusions in patients with advanced non-small cell lung carcinomas using the VENTANA ROS1 (SP384) rabbit monoclonal primary antibody.

Expert Rev Mol Diagn 2021 Apr 26. Epub 2021 Apr 26.

Pathology and Laboratory of Therapeutic Targets, Hospital Universitario HM Sanchinarro, HM Hospitales, CIBERONC, Madrid, Spain.

: The development of several ROS1 inhibitors means that the importance of accurately identifying -positive lung cancer patients has never been greater. Therefore, it is crucial that testing assays become more standardized.: Based on primary literature, combined with personal diagnostic and research experience, this review provide a pragmatic update on the use of the recently released VENTANA ROS1 (SP384) Rabbit Monoclonal Primary Antibody.: This assay provides high sensitivity, so it is an excellent analytical option when screening for fusions in patients with advanced non-small cell lung carcinomas.
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http://dx.doi.org/10.1080/14737159.2021.1919512DOI Listing
April 2021

Molecular diagnosis in non-small-cell lung cancer: expert opinion on and testing.

J Clin Pathol 2021 Apr 19. Epub 2021 Apr 19.

Department of Pathology, Hospital Universitario Virgen del Rocío, Sevilla, Spain.

The effectiveness of targeted therapies with tyrosine kinase inhibitors in non-small-cell lung cancer (NSCLC) depends on the accurate determination of the genomic status of the tumour. For this reason, molecular analyses to detect genetic rearrangements in some genes (ie, , , ) have become standard in patients with advanced disease. Since immunohistochemistry is easier to implement and interpret, it is normally used as the screening procedure, while fluorescence in situ hybridisation (FISH) is used to confirm the rearrangement and decide on ambiguous immunostainings. Although FISH is considered the most sensitive method for the detection of and rearrangements, the interpretation of results requires detailed guidelines. In this review, we discuss the various technologies available to evaluate and genomic rearrangements using these techniques. Other techniques such as real-time PCR and next-generation sequencing have been developed recently to evaluate and gene rearrangements, but some limitations prevent their full implementation in the clinical setting. Similarly, liquid biopsies have the potential to change the treatment of patients with advanced lung cancer, but further research is required before this technology can be applied in routine clinical practice. We discuss the technical requirements of laboratories in the light of quality assurance programmes. Finally, we review the recent updates made to the guidelines for the determination of molecular biomarkers in patients with NSCLC.
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http://dx.doi.org/10.1136/jclinpath-2021-207490DOI Listing
April 2021

Immunotherapy Moves to the Early-Stage Setting in Non-Small Cell Lung Cancer: Emerging Evidence and the Role of Biomarkers.

Cancers (Basel) 2020 Nov 20;12(11). Epub 2020 Nov 20.

Department of Radiation Oncology, Hospital Universitario Quirónsalud Madrid, Pozuelo de Alarcón, 28223 Madrid, Spain.

Despite numerous advances in targeted therapy and immunotherapy in the last decade, lung cancer continues to present the highest mortality rate of all cancers. Targeted therapy based on specific genomic alterations, together with PD-1 and CTLA-4 axis blocking-based immunotherapy, have significantly improved survival in advanced non-small cell lung cancer (NSCLC) and both therapies are now well-established in this clinical setting. However, it is time for immunotherapy to be applied in patients with early-stage disease, which would be an important qualitative leap in the treatment of lung cancer patients with curative intent. Preliminary data from a multitude of studies are highly promising, but therapeutic decision-making should be guided by an understanding of the molecular features of the tumour and host. In the present review, we discuss the most recently published studies and ongoing clinical trials, controversies, future challenges and the role of biomarkers in the selection of best therapeutic options.
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http://dx.doi.org/10.3390/cancers12113459DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7699975PMC
November 2020

Pan-TRK Immunohistochemistry.

Arch Pathol Lab Med 2020 Oct 28. Epub 2020 Oct 28.

From Pathology and Laboratory of Therapeutic Targets, Hospital Universitario HM Sanchinarro, HMHospitales, CIBERONC, Madrid, Spain (Conde, Lopez-Rios).

Context.—: Food and Drug Administration-approved TRK inhibitors with impressive overall response rates are now available for patients with multiple cancer types that harbor NTRK rearrangements, yet the identification of NTRK fusions remains a difficult challenge. These alterations are highly recurrent in extremely rare malignancies or can be detected in exceedingly small subsets of common tumor types. A 2-step approach has been proposed, involving a screening by immunohistochemistry (IHC) followed by a confirmatory method (fluorescence in situ hybridization, reverse transcriptase-polymerase chain reaction, or next-generation sequencing) in cases expressing the protein. However, there is no interpretation guide for any of the available IHC clones.

Objective.—: To provide a pragmatic update on the use of pan-TRK IHC. Selected examples of the different IHC staining patterns across multiple histologies are shown.

Data Sources.—: Primary literature review with PubMed, combined with personal diagnostic and research experience.

Conclusions.—: In-depth knowledge of pan-TRK IHC will help pathologists implement a rational approach to the detection of NTRK fusions in human malignancies.
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http://dx.doi.org/10.5858/arpa.2020-0400-RADOI Listing
October 2020

Lung Cancer in Spain.

J Thorac Oncol 2021 02 24;16(2):197-204. Epub 2020 Oct 24.

Department of Thoracic Surgery, Hospital Universitari Mútua Terrassa, University of Barcelona, Terrassa, Barcelona, Spain; Lung Cancer Group, Network of Centers for Biomedical Research in Respiratory Diseases/Centro de Investigación Biomédica en Red Enfermedades Respiratorias, Terrassa, Barcelona, Spain.

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http://dx.doi.org/10.1016/j.jtho.2020.09.026DOI Listing
February 2021

A noninterventional, multinational study to assess PD-L1 expression in cytological and histological lung cancer specimens.

Cancer Cytopathol 2020 12 28;128(12):928-938. Epub 2020 Jul 28.

Department of Pathology, University of Aberdeen, Aberdeen, United Kingdom.

Background: The diagnosis of advanced lung cancer is made with minimally invasive procedures. This often results in the availability of cytological material only for subtype determination and companion diagnostic testing, with the latter being technically and clinically validated on histological material only. Thus, the primary objective of the MO29978 clinical study was to assess programmed death ligand 1 (PD-L1) protein expression on cytology samples as surrogates for histology samples in patients with lung cancer.

Methods: Formalin-fixed, paraffin-embedded histological samples and cytological cell blocks from 190 patients were analyzed with immunohistochemical assays using the rabbit monoclonal anti-PD-L1 antibody clones SP142 and SP263. PD-L1 expression was quantified on both tumor cells (TC) and tumor-infiltrating immune cells (IC). Overall concordance, sensitivity, specificity, and accuracy, with a 1% cutoff used for both assays, were assessed for PD-L1 expression on TC and IC.

Results: In non-small cell lung cancer histology and cytology samples measured with the PD-L1 (SP142) antibody (n = 173), the intraclass correlation coefficients were 0.40 and 0.06 on TC and IC, respectively. With SP142 and SP263, accuracies of 74.1% for TC and 51.9% for IC and accuracies of 75.2% for TC and 61.2% for IC, respectively, were reported.

Conclusions: Overall, this study has demonstrated that PD-L1 analysis on TC is feasible in cytological material, but quantification is challenging. Tumor tissue should be preferred over cell block cytology for PD-L1 immunohistochemical analysis unless laboratories have validated their cytology preanalytical approaches and demonstrated the comparability of histology and cytology for TC PD-L1 results.
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http://dx.doi.org/10.1002/cncy.22324DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7754298PMC
December 2020

[Updated guidelines for predictive biomarker testing in advanced non-small-cell lung cancer: A National Consensus of the Spanish Society of Pathology and the Spanish Society of Medical Oncology].

Rev Esp Patol 2020 Jul - Sep;53(3):167-181. Epub 2020 Jun 16.

Departamento de Oncología Médica, Hospital Universitario Ramón y Cajal, Universidad Alcalá, IRYCIS, CIBERONC, Madrid, España.

In 2011, the Spanish Society of Medical Oncology (SEOM) and the Spanish Society of Pathology (SEAP) initiated a joint project to establish guidelines for biomarker testing in patients with advanced non-small-cell lung cancer based on the information available at the time. As this field is constantly evolving, these guidelines were updated in 2012 and 2015 and now in 2019. Current evidence suggests it should be mandatory to test all patients with this kind of advanced lung cancer for EGFR and BRAF mutations, ALK and ROS1 rearrangements and PD-L1 expression. The growing need to study other emerging biomarkers has promoted the routine use of massive sequencing (next-generation sequencing, NGS). However, the coordination of every professional involved and the prioritisation of the most suitable tests and technologies for each case remain a challenge.
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http://dx.doi.org/10.1016/j.patol.2019.11.004DOI Listing
June 2020

Hydration in relation to water insecurity, heat index, and lactation status in two small-scale populations in hot-humid and hot-arid environments.

Am J Hum Biol 2021 01 24;33(1):e23447. Epub 2020 Jun 24.

Department of Biobehavioral Health, The Pennsylvania State University, University Park, Pennsylvania, USA.

Objectives: This study compared the prevalence of concentrated urine (urine specific gravity ≥1.021), an indicator of hypohydration, across Tsimane' hunter-forager-horticulturalists living in hot-humid lowland Bolivia and Daasanach agropastoralists living in hot-arid Northern Kenya. It tested the hypotheses that household water and food insecurity would be associated with higher odds of hypohydration.

Methods: This study collected spot urine samples and corresponding weather data along with data on household water and food insecurity, demographics, and health characteristics among 266 Tsimane' households (N = 224 men, 235 women, 219 children) and 136 Daasanach households (N = 107 men, 120 women, 102 children).

Results: The prevalence of hypohydration among Tsimane' men (50.0%) and women (54.0%) was substantially higher (P < .001) than for Daasanach men (15.9%) and women (17.5%); the prevalence of hypohydration among Tsimane' (37.0%) and Daasanach (31.4%) children was not significantly different (P = .33). Multiple logistic regression models suggested positive but not statistically significant trends between household water insecurity and odds of hypohydration within populations, yet some significant joint effects of water and food insecurity were observed. Heat index (2°C) was associated with a 23% (95% confidence interval [CI]: 1.09-1.40, P = .001), 34% (95% CI: 1.18-1.53, P < .0005), and 23% (95% CI: 1.04-1.44, P = .01) higher odds of hypohydration among Tsimane' men, women, and children, respectively, and a 48% (95% CI: 1.02-2.15, P = .04) increase in the odds among Daasanach women. Lactation status was also associated with hypohydration among Tsimane' women (odds ratio = 3.35, 95% CI: 1.62-6.95, P = .001).

Conclusion: These results suggest that heat stress and reproductive status may have a greater impact on hydration status than water insecurity across diverse ecological contexts.
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http://dx.doi.org/10.1002/ajhb.23447DOI Listing
January 2021

Assessment of a New ROS1 Immunohistochemistry Clone (SP384) for the Identification of ROS1 Rearrangements in Patients with Non-Small Cell Lung Carcinoma: the ROSING Study.

J Thorac Oncol 2019 12 23;14(12):2120-2132. Epub 2019 Jul 23.

Alvaro Cunqueiro Hospital, Vigo, Spain.

Introduction: The ROS1 gene rearrangement has become an important biomarker in NSCLC. The College of American Pathologists/International Association for the Study of Lung Cancer/Association for Molecular Pathology testing guidelines support the use of ROS1 immunohistochemistry (IHC) as a screening test, followed by confirmation with fluorescence in situ hybridization (FISH) or a molecular test in all positive results. We have evaluated a novel anti-ROS1 IHC antibody (SP384) in a large multicenter series to obtain real-world data.

Methods: A total of 43 ROS1 FISH-positive and 193 ROS1 FISH-negative NSCLC samples were studied. All specimens were screened by using two antibodies (clone D4D6 from Cell Signaling Technology and clone SP384 from Ventana Medical Systems), and the different interpretation criteria were compared with break-apart FISH (Vysis). FISH-positive samples were also analyzed with next-generation sequencing (Oncomine Dx Target Test Panel, Thermo Fisher Scientific).

Results: An H-score of 150 or higher or the presence of at least 70% of tumor cells with an intensity of staining of 2+ or higher by the SP384 clone was the optimal cutoff value (both with 93% sensitivity and 100% specificity). The D4D6 clone showed similar results, with an H-score of at least 100 (91% sensitivity and 100% specificity). ROS1 expression in normal lung was more frequent with use of the SP384 clone (p < 0.0001). The ezrin gene (EZR)-ROS1 variant was associated with membranous staining and an isolated green signal FISH pattern (p = 0.001 and p = 0.017, respectively).

Conclusions: The new SP384 ROS1 IHC clone showed excellent sensitivity without compromising specificity, so it is another excellent analytical option for the proposed testing algorithm.
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http://dx.doi.org/10.1016/j.jtho.2019.07.005DOI Listing
December 2019

FGFR1 Cooperates with EGFR in Lung Cancer Oncogenesis, and Their Combined Inhibition Shows Improved Efficacy.

J Thorac Oncol 2019 04 9;14(4):641-655. Epub 2019 Jan 9.

H12O-CNIO Lung Cancer Clinical Research Unit, Biomedical Research Foundation i+12, Madrid, Spain; H12O-CNIO Lung Cancer Clinical Research Unit, Spanish National Cancer Research Centre (CNIO), Madrid, Spain; CIBERONC, Madrid, Spain; Medical Oncology Department, University Hospital Doce de Octubre Madrid, Spain; Medical School, Complutense University, Madrid, Spain.

Introduction: There is substantial evidence for the oncogenic effects of fibroblast growth factor receptor 1 (FGFR1) in many types of cancer, including lung cancer, but the role of this receptor has not been addressed specifically in lung adenocarcinoma.

Methods: We performed FGFR1 and EGFR overexpression and co-overexpression assays in adenocarcinoma and in inmortalized lung cell lines, and we also carried out surrogate and interaction assays. We performed monotherapy and combination EGFR/FGFR inhibitor sensitivity assays in vitro and in vivo in cell line- and patient-derived xenografts. We determined FGFR1 mRNA expression in a cohort of patients with anti-EGFR therapy-treated adenocarcinoma.

Results: We have reported a cooperative interaction between FGFR1 and EGFR in this context, resulting in increased EGFR activation and oncogenic signaling. We have provided in vitro and in vivo evidence indicating that FGFR1 expression increases tumorigenicity in cells with high EGFR activation in EGFR-mutated and EGFR wild-type models. At the clinical level, we have shown that high FGFR1 expression levels predict higher resistance to erlotinib or gefitinib in a cohort of patients with tyrosine kinase inhibitor-treated EGFR-mutated and EGFR wild-type lung adenocarcinoma. Dual EGFR and FGFR inhibition in FGFR1-overexpressing, EGFR-activated models shows synergistic effects on tumor growth in vitro and in cell line- and patient-derived xenografts, suggesting that patients with tumors bearing these characteristics may benefit from combined EGFR/FGFR inhibition.

Conclusion: These results support the extended the use of EGFR inhibitors beyond monotherapy in the EGFR-mutated adenocarcinoma setting in combination with FGFR inhibitors for selected patients with increased FGFR1 overexpression and EGFR activation.
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http://dx.doi.org/10.1016/j.jtho.2018.12.021DOI Listing
April 2019

Author Correction: Free mate choice does not influence reproductive success in humans.

Sci Rep 2018 Jul 3;8(1):10295. Epub 2018 Jul 3.

Department of Human Biology, University of Wroclaw, Wroclaw, Poland.

A correction to this article has been published and is linked from the HTML and PDF versions of this paper. The error has not been fixed in the paper.
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http://dx.doi.org/10.1038/s41598-018-28336-7DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6028642PMC
July 2018

Dental enamel defects predict adolescent health indicators: A cohort study among the Tsimane' of Bolivia.

Am J Hum Biol 2018 05 5;30(3):e23107. Epub 2018 Feb 5.

School of Dentistry, Department of Oral Health Sciences, University of Washington, Seattle, Washington, 98119.

Objectives: Bioarchaeological findings have linked defective enamel formation in preadulthood with adult mortality. We investigated how defective enamel formation in infancy and childhood is associated with risk factors for adult morbidity and mortality in adolescents.

Methods: This cohort study of 349 Amerindian adolescents (10-17 years of age) related extent of enamel defects on the central maxillary incisors (none, less than 1/3, 1/3 to 2/3, more than 2/3) to adolescent anthropometrics (height, weight) and biomarkers (hemoglobin, glycated hemoglobin, white blood cell count, and blood pressure). Risk differences and 95% confidence intervals were estimated using multiple linear regression. Enamel defects and stunted growth were compared in their ability to predict adolescent health indicators using log-binomial regression and receiver operating characteristics (ROCs).

Results: Greater extent of defective enamel formation on the tooth surface was associated with shorter height (-1.35 cm, 95% CI: -2.17, -0.53), lower weight (-0.98 kg, 95% CI: -1.70, -0.26), lower hemoglobin (-0.36 g/dL, 95% CI: -0.59, -0.13), lower glycated hemoglobin (-0.04 %A , 95% CI: -0.08, -0.00008), and higher white blood cell count (0.74 10 /L, 95% CI: 0.35, 1.14) in adolescence. Extent of enamel defects and stunted growth independently performed similarly as risk factors for adverse adolescent outcomes, including anemia, prediabetes/type II diabetes, elevated WBC count, prehypertension/hypertension, and metabolic health.

Conclusions: Defective enamel formation in infancy and childhood predicted adolescent health outcomes and may be primarily associated with infection. Extent of enamel defects and stunted growth may be equally predictive of adverse adolescent health outcomes.
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http://dx.doi.org/10.1002/ajhb.23107DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5980689PMC
May 2018

Free mate choice does not influence reproductive success in humans.

Sci Rep 2017 08 31;7(1):10127. Epub 2017 Aug 31.

Department of Human Biology, University of Wroclaw, Wroclaw, Poland.

The effect of free mate choice on the relative magnitude of fitness benefits has been examined among various species. The majority of the data show significant fitness benefits of mating with partners of an individual's own choice, highlighting elevated behavioral compatibility between partners with free mate choice. Similarities between humans and other species that benefit from free mate choice led us to hypothesize that it also confers reproductive benefits in Homo sapiens. To test this hypothesis, we conducted a study among three indigenous societies-the Tsimane', Yali, and Bhotiya-who employ natural birth control. In all three samples, we compared the marriages arranged by parents with the non-arranged ones in terms of number of offspring. Here, we show that there were no significant relationships between type of marriage and the total number of alive children and number of dead children among the three sampled groups. The presented study is the first to date to examine the fitness benefits of free mate choice in humans. In discussion we present limitations of our research and discuss the possibility of love having a beneficial influence in terms of the number of offspring.
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http://dx.doi.org/10.1038/s41598-017-10484-xDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5578983PMC
August 2017

Aligning digital CD8 scoring and targeted next-generation sequencing with programmed death ligand 1 expression: a pragmatic approach in early-stage squamous cell lung carcinoma.

Histopathology 2018 Jan 3;72(2):270-284. Epub 2017 Nov 3.

Pathology-Laboratorio de Dianas Terapeuticas, Hospital Universitario HM Sanchinarro, Universidad CEU San Pablo, Madrid, Spain.

Aims: To study programmed death ligand 1 (PD-L1) expression, tumour-infiltrating T lymphocytes (TILs) and the molecular context in patients with early-stage squamous cell lung carcinomas (SCCs).

Methods And Results: The study included samples from 40 patients (discovery cohort) and 29 patients (validation cohort) diagnosed with early-stage SCC. PD-L1 immunohistochemistry (IHC) was performed with three commercially available clones (E1L3N, SP263 and SP142). CD8 TILs were scored with a digital algorithm. All tumours were analysed with targeted next-generation sequencing (NGS). Additionally, TP53 mutations were investigated with direct sequencing. In both cohorts, we observed a significant association between CD8 TILs density and high PD-L1 IHC expression in tumour cells (TCs). Furthermore, high SP142 PD-L1 expression in immune cells (ICs) was also associated significantly with CD8 TILs density. Therefore, CD8 TILs density discriminated between patients with high versus low PD-L1 IHC expression with excellent sensitivity and specificity. Interestingly, the highest percentages of PD-L1-positive TCs with the three antibodies were found in samples with cyclin-dependent kinase 6 (CDK6) amplification, with high amplification of proto-oncogene C-Myc (CMYC) or with cyclin D1-PI3 kinase subunit alpha (CCND1-PIK3CA) co-amplification. High SP142 PD-L1 IHC expression in ICs showed a non-significant correlation with TP53 mutations. Conversely, most cases with fibroblast growth factor receptor 1 (FGFR1) amplification were negative for all PD-L1 clones.

Conclusions: Our preliminary results support the use of digital CD8 TILs scoring and targeted NGS alongside PD-L1 expression. The approach presented herein could help define patients with SCCs candidates to immune checkpoints inhibitors.
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http://dx.doi.org/10.1111/his.13346DOI Listing
January 2018

Malnutrition-related early childhood exposures and enamel defects in the permanent dentition: A longitudinal study from the Bolivian Amazon.

Am J Phys Anthropol 2017 10 28;164(2):416-423. Epub 2017 Jul 28.

School of Dentistry, Department of Oral Health Sciences, University of Washington, Seattle, Washington, DC, 98119.

Objectives: We investigated the relationship between early childhood malnutrition-related measures and subsequent enamel defects in the permanent dentition.

Materials And Methods: This cohort study included 349 Amerindian adolescents (10-17 years, 52% male) from the Bolivian Amazon. Exposures included: stunted growth (height-for-age z-scores), underweight (weight-for-age z-scores), anemia (hemoglobin), acute inflammation (C-reactive protein) and parasitic infection (hookworm). We measured the occurrence (no/yes) and extent (<1/3, 1/3-2/3, >2/3) of enamel defects. We estimated associations between childhood exposures and enamel defect measures using log-binomial and multinomial logistic regression.

Results: The prevalence of an enamel defect characterized by an orange peel texture on a large central depression on the labial surface of the central maxillary incisors was 92.3%. During childhood (1-4 years), participants had a high prevalence of stunted growth (75.2%), anemia (56.9%), acute inflammation (39.1%), and hookworm infection (49.6%). We observed associations between childhood height-for-age (OR = 0.65; P = 0.028 for >2/3 extent vs. no EH) and gastrointestinal hookworm infection (OR = 3.43; P = 0.035 for >2/3 extent vs. no defects or <1/3 extent) with enamel defects.

Discussion: The study describes a possibly novel form of enamel hypoplasia and provides evidence for associations of malnutrition-related measures in early childhood, including stunted growth and parasitic helminth infection, with the observed enamel defects.
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http://dx.doi.org/10.1002/ajpa.23283DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5607097PMC
October 2017

Identification of , , and Fusions by a Multiplexed mRNA-Based Assay in Formalin-Fixed, Paraffin-Embedded Samples from Advanced Non-Small-Cell Lung Cancer Patients.

Clin Chem 2017 03 10;63(3):751-760. Epub 2017 Jan 10.

Medical Oncology, Hospital Clínic, Barcelona, Spain.

Background: Anaplastic lymphoma receptor tyrosine kinase (), ROS proto-oncogene 1, receptor tyrosine kinase (), and ret proto-oncogene () fusions are present in 5%-7% of patients with advanced non-small-cell lung cancer (NSCLC); their accurate identification is critical to guide targeted therapies. FISH and immunohistochemistry (IHC) are considered the gold standards to determine gene fusions, but they have limitations. The nCounter platform is a potentially useful genomic tool for multiplexed detection of gene fusions, but has not been validated in the clinical setting.

Methods: Formalin-fixed, paraffin embedded (FFPE) samples from 108 patients with advanced NSCLC were analyzed with an nCounter-based assay and the results compared with FISH, IHC, and reverse transcription PCR (RT-PCR). Data on response to fusion kinase inhibitors was retrospectively collected in a subset of 29 patients.

Results: Of 108 FFPE samples, 98 were successfully analyzed by nCounter (91%), which identified 55 fusion-positive cases (32 , 21 , and 2 ). nCounter results were highly concordant with IHC for ALK (98.5%, CI = 91.8-99.7), while 11 discrepancies were found compared with FISH (87.5% concordance, CI = 79.0-92.9). For , nCounter showed similar agreement with IHC and FISH (87.2% and 85.9%), but a substantial number of samples were positive only by 1 or 2 techniques. Of the 25 patients deriving clinical benefit from fusion kinase inhibitors, 24 were positive by nCounter and 22 by FISH.

Conclusions: nCounter compares favorably with IHC and FISH and can be used for identifying patients with advanced NSCLC positive for // fusion genes.
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http://dx.doi.org/10.1373/clinchem.2016.265314DOI Listing
March 2017

Profile of Ventana ALK (D5F3) companion diagnostic assay for non-small-cell lung carcinomas.

Expert Rev Mol Diagn 2016 06 15;16(6):707-13. Epub 2016 Apr 15.

a Laboratorio de Dianas Terapéuticas , Hospital Universitario HM Sanchinarro , Madrid , Spain.

The development of several ALK inhibitors means that the importance of accurately identifying ALK-positive lung cancer has never been greater. Therefore, it is crucial that ALK testing assays become more standardized. The aim of this review is to comment on the recently FDA-approved VENTANA ALK (D5F3) Companion Diagnostic (CDx) Assay. This kit provides high sensitivity and specificity for the detection of ALK rearrangements and seamless integration into the laboratory workflow, with a fully automated analytical phase and fast interpretation. The use of controls increases the sensitivity and specificity and a dichotomous scoring approach enhances reproducibility.
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http://dx.doi.org/10.1586/14737159.2016.1172963DOI Listing
June 2016

Primary Cutaneous Ewing Sarcoma: Report of a Case.

Fetal Pediatr Pathol 2015 24;34(5):315-21. Epub 2015 Jul 24.

c Hospital Infantil Universitario Niño Jesús , Onco-Hematology , Madrid , Spain.

Primary cutaneous Ewing's sarcoma is a rare entity. Although the diagnosis may be very difficult, it can be confirmed through molecular biology. We present the case of a 13-years old male with a lesion in the sole of the right foot, characterized by a monomorphous proliferation of small, round and blue cells. The histology and molecular biology allowed us to perform the diagnosis of cutaneous Ewing's sarcoma. This neoplasm must be distinguished from other round cell tumors with cutaneous involvement. The prognosis and treatment of this rare disease will also be discussed.
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http://dx.doi.org/10.3109/15513815.2015.1068411DOI Listing
June 2016

The anaplastic lymphoma kinase testing conundrum.

Expert Rev Mol Diagn 2015 Feb 12;15(2):161-3. Epub 2015 Jan 12.

Laboratorio de Dianas Terapéuticas, Hospital HM Universitario Sanchinarro, Universidad San Pablo-CEU, Faculty of Medicine, C/Oña, 10. 28050 Madrid, Spain.

Given the excellent results of the clinical trials with anaplastic lymphoma kinase (ALK) inhibitors, the importance of accurately identifying ALK-positive lung carcinoma patients has never been greater. It brings with it a pressing need for harmonized development of companion diagnostics, for economic, scientific and medical reasons. Therefore, it is crucial that ALK testing assays become more standardized both in performance (analytical phase) and interpretation (post-analytical phase). We find that both methods currently recommended by College of American Pathologists/International Association for the Study of Lung Cancer/Association for Molecular Pathology guidelines (FISH and Immunohistochemistry) are reasonable approaches for primary routine ALK testing, if at least 50 tumor cells are scored and protocols are strictly followed. Moreover, due to the high demand to study multiple predictive biomarkers on different assay platforms, quick and reliable approaches to achieve this are essential to guide treatment decisions.
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http://dx.doi.org/10.1586/14737159.2015.997713DOI Listing
February 2015

Accurate identification of ALK positive lung carcinoma patients: novel FDA-cleared automated fluorescence in situ hybridization scanning system and ultrasensitive immunohistochemistry.

PLoS One 2014 23;9(9):e107200. Epub 2014 Sep 23.

Laboratorio de Dianas Terapéuticas, Centro Integral Oncológico "Clara Campal", Hospital Universitario Madrid Sanchinarro, Universidad San Pablo-CEU, Madrid, Spain.

Background: Based on the excellent results of the clinical trials with ALK-inhibitors, the importance of accurately identifying ALK positive lung cancer has never been greater. However, there are increasing number of recent publications addressing discordances between FISH and IHC. The controversy is further fuelled by the different regulatory approvals. This situation prompted us to investigate two ALK IHC antibodies (using a novel ultrasensitive detection-amplification kit) and an automated ALK FISH scanning system (FDA-cleared) in a series of non-small cell lung cancer tumor samples.

Methods: Forty-seven ALK FISH-positive and 56 ALK FISH-negative NSCLC samples were studied. All specimens were screened for ALK expression by two IHC antibodies (clone 5A4 from Novocastra and clone D5F3 from Ventana) and for ALK rearrangement by FISH (Vysis ALK FISH break-apart kit), which was automatically captured and scored by using Bioview's automated scanning system.

Results: All positive cases with the IHC antibodies were FISH-positive. There was only one IHC-negative case with both antibodies which showed a FISH-positive result. The overall sensitivity and specificity of the IHC in comparison with FISH were 98% and 100%, respectively.

Conclusions: The specificity of these ultrasensitive IHC assays may obviate the need for FISH confirmation in positive IHC cases. However, the likelihood of false negative IHC results strengthens the case for FISH testing, at least in some situations.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0107200PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4172507PMC
June 2015

Tuberculous dacryoadenitis unveils HIV infection.

Can J Ophthalmol 2013 Oct;48(5):e128-30

Hospital Universitario de Fuenlabrada. Electronic address:

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http://dx.doi.org/10.1016/j.jcjo.2013.05.008DOI Listing
October 2013

Determining the profiles and parameters for gene amplification testing of growth factor receptors in lung cancer.

Int J Cancer 2013 Aug 9;133(4):898-907. Epub 2013 Mar 9.

Genes and Cancer Group, Cancer Epigenetics and Biology Program (PEBC), Bellvitge Biomedical Research Institute (IDIBELL), Hospitalet de Llobregat, Barcelona, Spain.

Growth factor receptors (GFRs) are amenable to therapeutic intervention in cancer and it is important to select patients appropriately. One of the mechanisms for activation of GFRs is gene amplification (GA) but discrepancies arising from the difficulties associated with data interpretation and the lack of agreed parameters confound the comparison of results from different laboratories. Here, we attempt to establish appropriate conditions for standardization of the determination of GA in a panel of GFRs. A NSCLC tissue microarray panel containing 302 samples was screened for alterations at ALK, FGFR1, FGFR2, FGFR3, ERBB2, IGF1R, KIT, MET and PDGFRA by FISH, immunostaining and/or real-time quantitative RT-PCR. Strong amplification was found for FGFR1, ERBB2, KIT/PDFGRA and MET, with frequencies ranging from 1 to 6%. Thresholds for overexpression and GA were established. Strong immunostaining was found in most tumors with ERBB2, MET and KIT amplification, although some tumors underwent strong immunostaining in the absence of GA. KIT and PDFGRA were always coamplified, but only one tumor showed PDGFRA overexpression, indicating that KIT is the main target. Amplification of FGFR1 predominated in squamous cell carcinomas, although the association with overexpression was inconclusive. Interestingly, alterations at ALK, MET, EGFR, ERBB2 and KRAS correlated with augmented levels of phospho-S6 protein, suggesting activation of the mTOR pathway, which may prove useful to pre-select tumors for testing. Overall, here, we provide with parameters for the determination of GA at ERBB2, MET, KIT and PDGFRA which could be implemented in the clinic to stratify lung cancer patients for specific treatments.
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http://dx.doi.org/10.1002/ijc.28090DOI Listing
August 2013

Comparison of molecular testing methods for the detection of EGFR mutations in formalin-fixed paraffin-embedded tissue specimens of non-small cell lung cancer.

J Clin Pathol 2013 May 5;66(5):381-5. Epub 2013 Feb 5.

Laboratorio de Dianas Terapeuticas, Centro Integral Oncologico Clara Campal, Hospital Universitario Madrid Sanchinarro, Madrid, Spain.

Aim: To conduct a methods correlation study of three different assays for the detection of mutations at EGFR gene in human formalin-fixed paraffin-embedded tumour (FFPET) specimens of non-small cell lung carcinomas (NSCLC).

Methods: We conducted a 2-site method comparison study of two european conformity (CE) in vitro diagnostic (IVD)-marked assays, the cobas EGFR Mutation Test and the Therascreen EGFR29 Mutation Kit, and 2× bidirectional Sanger sequencing. We blind-tested 124 NSCLC FFPET specimens with all three methods; the cobas test was performed at both sites. Positive (PPA) and negative percent agreements (NPA) were determined for the cobas test versus each of the other two methods. Specimens yielding discordant test results between methods were further tested using quantitative massively parallel pyrosequencing (MPP).

Results: PPA between cobas and Sanger was 98.8%; NPA was 79.3%. Overall there were seven discordant results. MPP confirmed an exon 19 deletion in two cases and L858R mutation in four cases. PPA between cobas and Therascreen was 98.9% and NPA was 100%. There was one discordant result. Reproducibility of the cobas test between the two sites was 99.2%.

Conclusions: The invalid rates for the cobas test and Therascreen were lower than Sanger sequencing. The cobas and Therascreen assays showed a high degree of concordance, and both were more sensitive for the detection of exon 19 deletion and L858R mutations than Sanger. The cobas test was highly reproducible between the two testing sites, used the least amount of DNA input and was the only test with automated results reporting.
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http://dx.doi.org/10.1136/jclinpath-2012-201240DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3632986PMC
May 2013

The ALK translocation in advanced non-small-cell lung carcinomas: preapproval testing experience at a single cancer centre.

Histopathology 2013 Mar 5;62(4):609-16. Epub 2013 Feb 5.

Laboratorio de Dianas Terapéuticas, Centro Integral Oncológico Clara Campal, Hospital Universitario Madrid Sanchinarro, Universidad San Pablo-CEU, Madrid, Spain.

Aims: To study the ALK translocation in patients with advanced non-small-cell lung carcinomas (NSCLCs) seen at a European cancer centre, and its association with EGFR mutations, KRAS mutations and MET amplification.

Methods And Results: The study included samples from 86 patients diagnosed with advanced NSCLC. ALK fluorescence in-situ hybridization (FISH) was performed using the ALK break-apart probe set (Vysis). ALK FISH-positive cases were defined as those with more than 15% break-apart signals or isolated red signals in 50 cells. EGFR and KRAS mutations were determined by direct sequencing. All ALK-positive cases were analysed retrospectively for MET amplification using a FISH assay, and for ALK mutations by sequencing. We found nine (10.5%) ALK-positive cases, all in adenocarcinomas and the majority in female patients (88.9%). Signet ring cells were observed in four (44.4%) of the nine patients. None of the ALK translocated cases showed MET amplifications or EGFR, KRAS and ALK mutations.

Conclusions: The prevalence of ALK translocation in an unselected population of European patients with advanced NSCLCs was 10%. This alteration was mutually exclusive with EGFR and KRAS mutations, as well as with MET amplification. If multiplexing is considered at the preanalytical phase, lung biopsy specimens are sufficient for performing several predictive assays.
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http://dx.doi.org/10.1111/his.12037DOI Listing
March 2013

Diplopia from pleomorphic lipoma of the orbit with lateral rectus muscle involvement.

Ophthalmic Plast Reconstr Surg 2013 Mar-Apr;29(2):e53-5

Hospital Universitario de Fuenlabrada, Madrid, Spain.

A-55-year-old man with a 2-year history of left proptosis with painless swelling of the upper and lateral bulbar conjunctiva was referred. He had developed diplopia in left gaze. Orbital CT showed left proptosis with a mass measuring 2 × 1 cm in the superolateral and lateral left orbit, with lateral rectus muscle infiltration. The lesion was excised and was found to be diffuse, and an infiltrative mass affecting the anterior portion of the lateral rectus muscle was also removed. The histopathologic diagnosis was pleomorphic lipoma. Only 7 cases of pleomorphic lipomas occurring in ocular adnexal tissues or in the orbit have been previously reported, but in none of the cases had an infiltration of the lateral rectus muscle or diplopia been described before. The histopathologic features and differential diagnosis of this type of soft tissue tumor are also described.
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http://dx.doi.org/10.1097/IOP.0b013e31826a5112DOI Listing
August 2013

Comparison of testing methods for the detection of BRAF V600E mutations in malignant melanoma: pre-approval validation study of the companion diagnostic test for vemurafenib.

PLoS One 2013 10;8(1):e53733. Epub 2013 Jan 10.

Laboratorio de Dianas Terapéuticas, Hospital Universitario Sanchinarro, Madrid, Spain.

Background: The cobas 4800 BRAF V600 Mutation Test is a CE-marked and FDA-approved in vitro diagnostic assay used to select patients with metastatic melanoma for treatment with the selective BRAF inhibitor vemurafenib. We describe the pre-approval validation of this test in two external laboratories.

Methods: Melanoma specimens were tested for BRAF V600 mutations at two laboratories with the: cobas BRAF Mutation Test; ABI BRAF test; and bidirectional direct sequencing. Positive (PPA) and negative (NPA) percent agreements were determined between the cobas test and the other assays. Specimens with discordant results were tested with massively parallel pyrosequencing (454). DNA blends with 5% mutant alleles were tested to assess detection rates.

Results: Invalid results were observed in 8/116 specimens (6·9%) with Sanger, 10/116 (8·6%) with ABI BRAF, and 0/232 (0%) with the cobas BRAF test. PPA was 97·7% for V600E mutation for the cobas BRAF test and Sanger, and NPA was 95·3%. For the cobas BRAF test and ABI BRAF, PPA was 71·9% and NPA 83·7%. For 16 cobas BRAF test-negative/ABI BRAF-positive specimens, 454 sequencing detected no codon 600 mutations in 12 and variant codon 600 mutations in four. For eight cobas BRAF test-positive/ABI BRAF-negative specimens, four were V600E and four V600K by 454 sequencing. Detection rates for 5% mutation blends were 100% for the cobas BRAF test, 33% for Sanger, and 21% for the ABI BRAF. Reproducibility of the cobas BRAF test was 111/116 (96%) between the two sites.

Conclusions: It is feasible to evaluate potential companion diagnostic tests in external laboratories simultaneously to the pivotal clinical trial validation. The health authority approved assay had substantially better performance characteristics than the two other methods. The overall success of the cobas BRAF test is a proof of concept for future biomarker development.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0053733PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3542342PMC
July 2013

A comparison of EGFR mutation testing methods in lung carcinoma: direct sequencing, real-time PCR and immunohistochemistry.

PLoS One 2012 27;7(8):e43842. Epub 2012 Aug 27.

Laboratorio de Dianas Terapéuticas, Faculty of Medicine, Centro Integral Oncológico Clara Campal, Hospital HM Universitario Sanchinarro, Universidad San Pablo-CEU, Madrid, Spain.

The objective of this study is to compare two EGFR testing methodologies (a commercial real-time PCR kit and a specific EGFR mutant immunohistochemistry), with direct sequencing and to investigate the limit of detection (LOD) of both PCR-based methods. We identified EGFR mutations in 21 (16%) of the 136 tumours analyzed by direct sequencing. Interestingly, the Therascreen EGFR Mutation Test kit was able to characterize as wild-type one tumour that could not be analyzed by direct sequencing of the PCR product. We then compared the LOD of the kit and that of direct sequencing using the available mutant tumours. The kit was able to detect the presence of a mutation in a 1% dilution of the total DNA in nine of the 18 tumours (50%), which tested positive with the real-time quantitative PCR method. In all cases, EGFR mutation was identified at a dilution of 5%. Where the mutant DNA represented 30% of the total DNA, sequencing was able to detect mutations in 12 out of 19 cases (63%). Additional experiments with genetically defined standards (EGFR ΔE746-A750/+ and EGFR L858R/+) yielded similar results. Immunohistochemistry (IHC) staining with exon 19-specific antibody was seen in eight out of nine cases with E746-A750del detected by direct sequencing. Neither of the two tumours with complex deletions were positive. Of the five L858R-mutated tumours detected by the PCR methods, only two were positive for the exon 21-specific antibody. The specificity was 100% for both antibodies. The LOD of the real-time PCR method was lower than that of direct sequencing. The mutation specific IHC produced excellent specificity.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0043842PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3428292PMC
February 2013

Epiphyseal ewing sarcoma: first reported case with molecular confirmation.

Int J Surg Pathol 2013 Apr 25;21(2):173-6. Epub 2012 Jul 25.

Department of Surgical Pathology, Hospital Universitario de Móstoles, Madrid, Spain.

Ewing sarcoma is the second most common pediatric malignant bone neoplasm after osteosarcoma. Ewing sarcoma comprises "small, round, blue-cell" tumors thought to arise from neural crest cells. The authors report the case of a 14-year-old boy that presented with a nonpainful circumscribed lesion. The radiographs showed a lytic lesion at the tibial epiphysis with a large soft tissue mass, best depicted in the magnetic resonance imaging scan that suggested an aggressive lesion. A needle biopsy of the lesion was performed. The diagnosis of Ewing sarcoma was made based on microscopic, immunohistochemical, polymerase chain reaction, and fluorescence in situ hybridization. This is the third case report about a primary epiphyseal Ewing sarcoma and the fist one with molecular confirmation.
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http://dx.doi.org/10.1177/1066896912453202DOI Listing
April 2013

Chromophobe renal cell carcinoma: a review of an uncommon entity.

Int J Urol 2012 Oct 21;19(10):894-900. Epub 2012 Jun 21.

Department of Medical Oncology, Clara Campal Comprehensive Cancer Center, Madrid Sanchinarro University Hospital, Madrid, Spain.

Renal cell carcinoma is the most common neoplasm of the kidney. It is a heterogeneous disease, comprised of different histological variants with a distinct clinical course, genetics and response to treatment. The various subtypes identified include clear cell, papillary and chromophobe, among others. Chromophobe renal cell carcinoma is a rare variant and accounts for 5% of all cases. These tumors are macroscopically larger when compared with other forms and are commonly diagnosed at an early stage. Despite significant advances in renal cell carcinoma therapeutics in the past decade, no standard treatment has been identified for advanced chromophobe renal cell carcinoma. Nevertheless, new molecular insights have recently become available. A familial form of renal cell carcinoma, the Birt-Hogg-Dubé syndrome, has been described and the knowledge obtained has opened research opportunities in the therapeutic arena of chromophobe renal cell carcinoma. The following manuscript will endeavor to provide an overview of this uncommon entity including pathology, epidemiology, genetics, clinical aspects, and current and future treatment options.
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http://dx.doi.org/10.1111/j.1442-2042.2012.03079.xDOI Listing
October 2012