Silvia Hernández - Academia.edu (original) (raw)

Papers by Silvia Hernández

Research paper thumbnail of Prospective Study of FGFR3 Mutations As a Prognostic Factor in Nonmuscle Invasive Urothelial Bladder Carcinomas

Journal of Clinical Oncology, 2006

Purpose To determine the frequency and the prognostic value of fibroblast growth factor receptor ... more Purpose To determine the frequency and the prognostic value of fibroblast growth factor receptor 3 (FGFR3) mutations in patients with nonmuscle invasive bladder tumors according to tumor stage and grade. Patients and Methods Seven hundred seventy-two patients with newly diagnosed bladder tumors were recruited. Tumors were reviewed by expert pathologists. Patients were prospectively followed-up (median, 62.6 months for disease-free patients) through review of hospital records and telephone interviews. The sequence of exons 7 and 10 of FGFR3 was analyzed by polymerase chain reaction and direct sequencing. We assessed the association of mutations with stage and grade. The predictive value of mutations for recurrence, progression, and mortality were assessed using Kaplan-Meier and Cox multivariable models. Results Mutations were more common among low malignant potential neoplasms (LMPN; 77%) and TaG1/TaG2 tumors (61%/58%) than among TaG3 tumors (34%) and T1G3 tumors (17%). The S249C, Y3...

Research paper thumbnail of Clinical Interest of LMO2 Testing for the Diagnosis of Aggressive Large B-Cell Lymphomas

Cancers

MYC rearrangements usually confer aggressive biological behavior to large B-cell lymphomas. In th... more MYC rearrangements usually confer aggressive biological behavior to large B-cell lymphomas. In this study, we aimed to evaluate the relevance of LMO2 detection to the clinical approach to these tumors. First, the ability of LMO2 loss of expression to recognize the presence of MYC rearrangements was evaluated. A series of 365 samples obtained from 351 patients, including 28 Burkitt lymphoma, 230 diffuse large B-cell lymphoma, 30 high-grade B-cell lymphoma with MYC and BCL2/BCL6 rearrangements, eight high-grade B-cell lymphoma-NOS, 43 transformed diffuse large B-cell lymphoma, and 26 high-grade follicular lymphomas was analyzed. Among the CD10-positive tumors prospectively analyzed in whole tissue sections, LMO2 negative expression obtained values of 88% sensitivity, 94% specificity, and 93% accuracy, proving the utility of LMO2 to screen MYC rearrangements. In addition, survival analyses were performed in a series of 155 patients. As per univariate analyses, the prognosis relevance o...

Research paper thumbnail of Complementary value of electron microscopy and immunohistochemistry in the diagnosis of non-small cell lung cancer: A potential role for electron microscopy in the era of targeted therapy

Ultrastructural Pathology, 2019

With the identification of therapeutic targets for lung adenocarcinoma, it has become mandatory t... more With the identification of therapeutic targets for lung adenocarcinoma, it has become mandatory to distinguish it from other entities. Some cases remain classified as non-small cell lung carcinoma, not otherwise specified (NSCLC-NOS) with immunohistochemistry. Electron microscopy (EM) can be useful, allowing the identification of glandular differentiation. The aim of this study was to determine the complementary value of immunohistochemistry and EM. Forty-eight NSCLC-NOS cases were selected (PSMAR-Biobank, Barcelona, Spain). Immunohistochemistry (TTF-1, p40) was performed. Tissue was retrieved from paraffin blocks. Results were compared to the final diagnosis, derived from combination of light microscopy, immunohistochemistry, EM, molecular studies and resection specimen. Immunohistochemistry concurred with final diagnosis in 36 cases (75%, Kappa = 0.517). EM agreed with final diagnosis in 35 (72.9%, Kappa = 0.471). Immunohistochemistry had a sensitivity = 73%, specificity = 100%, positive predictive value (PPV) = 100% and negative predictive value (NPV) = 52.4% for adenocarcinoma. All adenocarcinoma cases not solved by immunohistochemistry (n = 10) were classified by EM, and vice versa. Data from EM were identical to those of immunohistochemistry: sensitivity = 73%, specificity = 100%, PPV = 100% and NPV = 52.4%. Combining both techniques, 47 cases were coincident with final diagnosis (97.9%, Kappa = 0.943). EM can provide valuable information in subtyping NSCLC-NOS, being particularly useful when immunohistochemistry is inconclusive. EM could be considered as a complementary tool for decision-making in NSCLC-NOS.

Research paper thumbnail of ERG overexpression plus SLC45A3 (prostein) and PTEN expression loss: Strong association of the triple hit phenotype with an aggressive pathway of prostate cancer progression

Oncotarget, Sep 26, 2017

TMPRSS2 and SLC45A3 rearrangements may coexist in the same tumor. ERG rearrangements and PTEN los... more TMPRSS2 and SLC45A3 rearrangements may coexist in the same tumor. ERG rearrangements and PTEN loss are concomitant events in prostate cancer (PrCa), and can cooperate in progression. We have reported that mRNA expression of TMPRSS2-ERG and SLC45A3-ERG rearrangements plus PTEN loss define an aggressive tumor subset. The aim of this study has been to validate these results by immunohistochemistry in a large cohort of tumors. ERG, SLC45A3 and PTEN immunostaining and their association with pathological features and PSA progression-free survival were analyzed in 220 PrCa (PSMAR-Biobank, Barcelona, Spain). ERG protein expression was found in 46.8% and SLC45A3 and PTEN loss in 30% and 34% tumors, respectively. Single ERG positive immunostaining was associated with GS = 6 tumors (p = 0.016), double ERG+/PTEN loss with GS = 7 (p = 0.008) and Grade Group 2 (GG) or GG3 cases (p = 0.042), ERG+/SLC45A3 loss/PTEN loss ("triple hit") with GS ≥ 8 (p < 0.0001) and GG4 or GG5 tumors (p =...

Research paper thumbnail of FOXO1 down-regulation is associated with worse outcome in bladder cancer and adds significant prognostic information to p53 overexpression

Research paper thumbnail of 803 PI3K signalling pathway is activated by PIK3CA gain and overexpression in prostate tumours, but PI3KCA, BRAF, KRAS AND AKT1 mutations are an infrequent event

European Journal of Cancer Supplements, 2010

tumourigenesis in this subgroup remain poorly described. We have analysed copy number aberrations... more tumourigenesis in this subgroup remain poorly described. We have analysed copy number aberrations and mutation profile of known target genes in tumours from early and late onset CRC patients. Material and Methods: High resolution array CGH analysis (385 000 features), was performed in 23 patients with CRC diagnosis at young age (range: 28−53 years, median: 44 years), and 17 patients with CRC diagnosis at old age (range: 69−78 years, median: 79 years). Furthermore, mutation profiles were analysed in a larger series of carcinomas stratified according to microsatellite instability analysis (MSI) status. These patients included 45 young-at-onset (range; 27−50 years, median; 43 years), and 69 old-atonset (n = 69, range; 71−93 years, median; 81 years). A panel of five genes, TP53, KRAS, BRAF, PTEN and PIK3CA, were investigated for mutations by sequencing. Results: The overall genome copy number profiles were similar between carcinomas from patients in the two age groups. However, some chromosomal stretches were found to have statistically significant (p < 0.05) more aberrations in the young patients compared to the old-at-onset group (not visa versa); DNA sequences within 2q, 10q, 19q, were more often gained, and sequences within 1p, 1q, 2q, 4p, 4q 10p and 19p, were more frequently lost. KRAS and PTEN mutations were distributed equally between the patient groups, whereas the mutation frequencies of TP53 and PIK3CA differed between the groups. BRAF mutations were not significantly correlated with MSI in the young-atonset group. Conclusions: We have identified genomic and gene specific differences in colorectal carcinomas related to time of disease onset. The somatic genomic changes that occur preferentially in tumours from young patients pinpoint potential genetic risk loci that will be further examined.

Research paper thumbnail of Association of ERG and TMPRSS2-ERG with grade, stage, and prognosis of prostate cancer is dependent on their expression levels

The Prostate, 2015

BACKGROUND. There is controversy in the literature on the role of the fusion TMPRSS2-ERG in the p... more BACKGROUND. There is controversy in the literature on the role of the fusion TMPRSS2-ERG in the pathogenesis and progression of prostate cancer. The quantitative differences in TMPRSS2-ERG fusion expression have received very limited attention in the literature. METHODS. We have quantitatively analyzed the mRNA levels of TMPRSS2-ERG, ERG, PTEN, and AR (n ¼ 83), as well as ERG immunostaining (n ¼ 78) in a series of prostate tumors. RESULTS. Among the TMPRSS2-ERG cases (n ¼ 57), high fusion levels were associated with GS !8 (P ¼ 0.025). ERG mRNA overexpression was associated with GS !8 (P ¼ 0.047), and with stage T3-T4 tumors (P ¼ 0.032). Among the ERG overexpressing cases (n ¼ 54), higher expression levels were found in 92.3% of GS !8 tumors (P ¼ 0.02). ERG immunostaining, regardless of staining intensity, was also associated with high stage (P ¼ 0.05). There was a statistical association between ERG immunostaining and PSA progression-free survival (Log Rank test, P ¼ 0.048). Decreased PTEN expression was associated with TMPRSS2-ERG (P ¼ 0.01), ERG mRNA overexpression (P ¼ 0.003) and ERG immunostaining (P ¼ 0.007). Furthermore, decreased PTEN expression, alone (P ¼ 0.041) and also combined with TMPRSS2-ERG (P ¼ 0.04) or with ERG overexpression (P ¼ 0.04) was associated with GS !7 tumors. CONCLUSIONS. Although more studies are needed to further clarify their role, our findings emphasize that the expression levels of the TMPRSS2-ERG fusion and ERG mRNA, rather than their mere presence, are related to a more aggressive phenotype, have an effect on prognosis and could be molecular markers of progression for prostate cancer. Furthermore, ERG immunohistochemistry could be also a potentially useful prognostic factor.

Research paper thumbnail of PI3K signaling pathway is activated by PIK3CA mRNA overexpression and copy gain in prostate tumors, but PIK3CA, BRAF, KRAS and AKT1 mutations are infrequent events

Modern Pathology, 2010

The phosphatidylinositol 3-kinase (PI3K)-AKT and RAS-MAPK pathways are deregulated in a wide rang... more The phosphatidylinositol 3-kinase (PI3K)-AKT and RAS-MAPK pathways are deregulated in a wide range of human cancers by gain or loss of function in several of their components. Our purpose has been to identify genetic alterations in members of these pathways in prostate cancer. A total of 102 prostate tumors, 79 from prostate cancer alone (group G1) and 23 from bladder and prostate cancer patients (G2), are the subject of this study. In 20 of these 23, the bladder tumors were also analyzed. PIK3CA, KRAS, BRAF and AKT1 mutations were analyzed by direct sequencing, and BRAF also by pyrosequencing. PIK3CA quantitative mRNA expression and fluorescence in situ hybridization (FISH) gains were tested in 25 and 32 prostate tumors from both groups (G1 and G2), respectively. Immunohistochemistry for pAKT was performed in 55 prostate tumors. Of 25 prostate tumors, 10 (40%) had PIK3CA mRNA overexpression that was statistically associated with Gleason score Z7 (P ¼ 0.018). PIK3CA copy gain was detected in 9 of 32 (28%) prostate tumors. Of 20 bladder tumors, 3 (15%) displayed mutations in PIK3CA, KRAS and AKT1, the corresponding prostate tumors being wt. We also detected a previously not reported PIK3CA polymorphism (IVS9 þ 91) in two prostate tumors. In all, 56% of prostate tumors overexpressed pAKT. There is a statistical association (Po0.0001) of strong pAKT immunostaining with high Gleason score, and with PIK3CA alterations (mRNA overexpression and/or FISH gains). PIK3CA gene is deregulated by mRNA overexpression and DNA gain in B40 and 28% of prostate tumors, respectively. High-grade prostate tumors are associated with PIK3CA mRNA overexpression, but not with FISH status. PIK3CA, BRAF, KRAS and AKT1 mutations are very infrequent events in prostate tumors. However, PI3K signaling pathway is activated by PIK3CA FISH gain and/or mRNA overexpression, leading to an increased pAKT protein expression.

Research paper thumbnail of PIK3CA Mutations Are an Early Genetic Alteration Associated with FGFR3 Mutations in Superficial Papillary Bladder Tumors

Cancer Research, 2006

Bladder tumors constitute a very heterogeneous disease. Superficial tumors are characterized by a... more Bladder tumors constitute a very heterogeneous disease. Superficial tumors are characterized by a high prevalence of FGFR3 mutations and chromosome 9 alterations. High-grade and muscle-invasive tumors are characterized by Tp53 mutations and aneuploidy. We have analyzed the sequence of exons 9 and 20 of PIK3CA in a panel of bladder tumors covering the whole spectrum of the disease. DNA from formalin-fixed, paraffin-embedded tumor sections was amplified by PCR and products were sequenced. In an unselected panel of tumors representative of the disease, the PIK3CA mutation prevalence was 13% (11 of 87). Mutations occurred mainly at the previously identified hotspots (codons 542, 545, 1007, and 1047). The distribution according to stage was as follows: papillary urothelial neoplasms of uncertain malignant potential (PUNLMP; 11 of 43, 25.6%), Ta (9 of 57, 16%), T1 (2 of 10, 20%), and muscle-invasive tumors (0 of 20, 0%; P = 0.019). Mutations were associated with low-grade tumors: grade 1 ...

Research paper thumbnail of Abstract 1168: Urothelial cell carcinomas harbor very frequent mutations in FGFR3-PI3K-AKT pathway and highest prevalence of mutations is related to low-grade tumors

Cancer Research, 2010

More than 90% of bladder cancers are urothelial cell carcinomas. Superficial/papillary and invasi... more More than 90% of bladder cancers are urothelial cell carcinomas. Superficial/papillary and invasive tumors progress via different molecular pathways. FGFR3 and PI3KCA mutations are more commonly found in superficial/papillary tumors. The PI3K-AKT pathway emerges as an alternative pathogenic mechanism linked to bladder cancer. We have analyzed the prevalence of mutations in FGFR3, PI3KCA, KRAS, BRAF and AKT1 genes in a set of 61 bladder tumors classified as TaG1 (n=10), TaG2 (n=19), TaG3 (n=8), T1G2 (n= 3), T1G3 (n= 11), T2G2 (n=1) and T2G3 (n=10). We focused on analyzing the hot spot mutation codons and surrounding regions. Mutation analysis of DNA from paraffin-embedded tumors was performed by direct sequencing of PCR products. Thirty-six (60%) tumors presented mutations, 22 cases with only one mutated gene, whereas 14 of 61 cases had mutations in two genes. The most frequently mutated gene was FGFR3 in 25 tumors, followed by PI3KCA in 16 tumors. The distribution and combinations o...

Research paper thumbnail of CXCR4 mRNA overexpression in high grade prostate tumors: lack of association with TMPRSS2-ERG rearrangement

The TMPRSS2-ERG fusion has been reported in 42 to 78% of prostate tumors. More than 90% of ERG-ov... more The TMPRSS2-ERG fusion has been reported in 42 to 78% of prostate tumors. More than 90% of ERG-overexpressing tumors harbor the fusion. The relationship between the TMPRSS2-ERG fusion and prognosis is controversial. Different studies have suggested an association between CXCR4 and ERG overexpression resulting from the TMPRSS2-ERG rearrangement. The aim of this study was to investigate the relationship between CXCR4 expression, TMPRSS2-ERG fusion and Gleason grade in prostate cancer. TMPRSS2-ERG rearrangement was investigated by FISH (n = 44), ERG protein by IHC (n = 84), and CXCR4 by quantitative RT-PCR (n = 44). TMPRSS2-ERG rearrangement and ERG protein expression were present in almost 50% of the cases, without statistical differences between the different Gleason score groups. There was a very high concordance between FISH and IHC techniques (Kappa Index = 0.954). Seventy percent of Gleason 8 prostate tumors overexpressed CXCR4 mRNA, and the difference in CXCR4 expression with Gleason < 8 cases was statistically significant (p = 0.009). There was no association between ERG protein and CXCR4 mRNA expression. In conclusion, our results reveal for the first time that CXCR4 overexpression is associated with high Gleason score prostate tumors, but that it is independent of the TMPRSS2-ERG rearrangement.

Research paper thumbnail of SPOP and CHD1 alterations in prostate cancer: Relationship with PTEN loss, tumor grade, perineural infiltration, and PSA recurrence

Research paper thumbnail of Immunohistochemical expression of mismatch repair proteins (MSH2, MSH6, MLH1, and PMS2) in prostate cancer: correlation with grade groups (WHO 2016) and ERG and PTEN status

Research paper thumbnail of SPOP and FOXA1 mutations are associated with PSA recurrence in ERG wt tumors, and SPOP downregulation with ERG ‐rearranged prostate cancer

Research paper thumbnail of Strong cytoplasmic ETV1 expression has a negative impact on prostate cancer outcome

Research paper thumbnail of Hernandez, S. et al. CDC25A and the splicing variant CDC25B2, but not CDC25B1, -B3 or -C, are over-expressed in aggressive human non-Hodgkin's lymphomas. Int. J. Cancer 89, 148-52

International Journal of Cancer

ABSTRACT

Research paper thumbnail of 816 FGFR3 mutations in prostate cancer and other tumours

European Journal of Cancer Supplements, 2010

Background: We previously reported FGFR3 mutations in prostate carcinoma (PCa). FGFR3 mutations w... more Background: We previously reported FGFR3 mutations in prostate carcinoma (PCa). FGFR3 mutations were associated with low-grade PCa, and also with PCa found in patients with concurrent bladder cancer or skin tumours. The aim of this work has been to further investigate the relationship between FGFR3 alterations in PCa and the presence of concurrent tumours. Material and Methods: 41 cases with PCa and other associated tumour types were studied. The PCa series consisted of: 22 incidental (cystoprostatectomy) tumours and 19 clinically significant (biopsy or prostatectomy) cases. Twentythree PCa were Gleason grade 6, 11 were Gleason grade 7 and 7 were Gleason grade 8. In each case, we studied the PCa and the concurrent tumour (prostate and bladder cancer, n = 32; prostate and skin tumour, n = 6; prostate and colon cancer, n = 2; prostate and lung cancer, n = 1). FGFR3 exons 7 and 10 were analysed by PCR and direct sequencing. Results: Eight of 41 (19.5%) PCa presented a mutation in FGFR3. From these, 6 were Gleason grade 6, and 2 were Gleason grade 7. Four of 32 (13%) patients with PCa and bladder cancer harboured a FGFR3 mutation in the PCa, and 5 other cases (16%) in the bladder tumour. In the PCa-skin tumour group, 3 of 6 (50%) PCa presented a FGFR3 mutation, and other 2 different cases (33%) in the skin tumour. One case harboured FGFR3 mutations in both tumours, but in different codons. Finally, one case with PCa and colon cancer also had a FGFR3 mutation in PCa. Conclusions: FGFR3 mutations in PCa are associated with an increased frequency of concurrent tumours in other organs, mainly skin and bladder. The lack of coincidence in the presence of FGFR3 mutations in both the PCa and the associated tumours suggests that they evolve through different pathways.

Research paper thumbnail of TRK-fused gene (TFG) is a new partner of ALK in anaplastic large cell lymphoma producing two structurally different TFG-ALK translocations

Blood, 1999

Anaplastic large cell lymphoma (ALCL) is associated with the t(2;5)(p23;q35), which generates the... more Anaplastic large cell lymphoma (ALCL) is associated with the t(2;5)(p23;q35), which generates the NPM-ALK fusion gene encoding an 80-kD protein. Several studies have suggested that genes other than NPM may be fused to the ALK gene. Here we have identified TRK-fused gene (TFG) as a new ALK partner in 2 ALCL, 1 of which exhibited a t(2;3)(p23;q21). In these cases, TFG was involved in 2 different fusion genes, TFG-ALK(S) and TFG-ALK(L), coding respectively 85-kD and 97-kD chimeric proteins. The ALK breakpoint in these translocations was the same as in the classic t(2;5) translocation. These 2 proteins were both active in an in vitro tyrosine kinase assay showing that the new cloned cDNA sequences are translated into chimeric proteins with functional activity. These findings indicate that TFG can provide an alternative to NPM as a fusion partner responsible for activation of the ALK and the pathogenesis of ALCL.

Research paper thumbnail of Mutations in FGFR3 and PIK3CA, singly or combined with RAS and AKT1, are associated with AKT but not with MAPK pathway activation in urothelial bladder cancer

Human Pathology, 2012

Different members of the phosphoinositide 3 kinase--serine threonine protein kinase (PI3K-AKT) pa... more Different members of the phosphoinositide 3 kinase--serine threonine protein kinase (PI3K-AKT) pathway are altered in bladder cancer. Fibroblast growth factor receptor 3 (FGFR3) mutations characterize the low-grade tumors, and RAS genes are mutated in approximately 13% of all bladder tumors. Interestingly, a percentage of bladder tumors have alterations in more than 1 PI3K-AKT or rat sarcoma viral oncogene homolog-RAF mitogen activated protein kinase (RAS-MAPK) pathway gene or their upstream regulators, but some combinations are mutually exclusive. We analyzed mutations in FGFR3, phosphoinositide 3 kinase catalytic alpha polypeptide (PIK3CA), v-akt murine thymoma viral oncogene homolog 1 (AKT1), v-Ki-ras2 Kirsten rat sarcoma viral oncogene homolog (KRAS), v-Ha-ras Harvey rat sarcoma viral oncogene homolog (HRAS), and v-raf murine sarcoma viral oncogene homolog B1 (BRAF) in 88 urothelial cell carcinomas and the immunohistochemical expression of phospho-v-akt murine thymoma viral oncogene homolog (AKT) and mitogen-activated protein kinase 1 and 2 (pERK1/2) in 80 and 77 urothelial cell carcinomas, respectively. Approximately 43% and 20.5% of tumors presented 1 and 2 mutated genes, respectively. FGFR3 mutations were more frequent alone, whereas PIK3CA mutations were associated with another mutated gene (FGFR3 and KRAS). Overall, mutated FGFR3 (FGFR3(mut)) and mutated FGFR3 (FGFR3(mut))-mutated PIK3CA (PIK3CA(mut)) genotypes were associated with low-grade bladder tumors and mutated PIK3CA (PIK3CA(mut))-mutated KRAS (KRAS(mut)) and mutated AKT1 (AKT1(mut)) were only present in high-grade tumors. There are no mutated FGFR3 (FGFR3(mut))-mutated RAS (RAS(mut)) nor mutated PIK3CA (PIK3CA(mut))-mutated AKT1 (AKT1(mut)) combinations. Fifty percent and 56% of tumors showed high levels of pAKT and pERK1/2, respectively. High levels of pAKT were associated with total mutations, FGFR3(mut), and PIK3CA(mut) tumors but not with tumor grade or stage. Wild-type tumors presented significantly higher pERK1/2 expression. Mutations in FGFR3 and FGFR3-PIK3CA but not single PIK3CA mutations characterize low-grade bladder tumors. Single FGFR3 or PIK3CA mutations and the different mutation combinations FGFR3-PIK3CA/AKT1 and PIK3CA-RAS can activate the AKT but not the MAPK pathway. Other genes different from FGFR3 may be related with the pERK activation in bladder tumors.

Research paper thumbnail of Manual Versus Laser Micro-dissection in Molecular Biology

Ultrastructural Pathology, 2006

Molecular studies on whole samples of fresh or frozen tissue do not take into account the heterog... more Molecular studies on whole samples of fresh or frozen tissue do not take into account the heterogeneity of these tissues. In addition to normal cells, precursor lesions and different progression stages may be mixed within a given sample. Usually, the dominant cell population will determine the results and may sometimes mask biologically relevant abnormalities. To obtain more specific information and knowledge on changes within different cell compartments, many techniques have been developed that combine morphological observation and selection with different strategies for specific cell dissection. In this review, the most important micro-dissection methods are put into perspective, and some requirements and limitations are discussed with regard to sample fixation, staining, dissection and molecular analysis.

Research paper thumbnail of Prospective Study of FGFR3 Mutations As a Prognostic Factor in Nonmuscle Invasive Urothelial Bladder Carcinomas

Journal of Clinical Oncology, 2006

Purpose To determine the frequency and the prognostic value of fibroblast growth factor receptor ... more Purpose To determine the frequency and the prognostic value of fibroblast growth factor receptor 3 (FGFR3) mutations in patients with nonmuscle invasive bladder tumors according to tumor stage and grade. Patients and Methods Seven hundred seventy-two patients with newly diagnosed bladder tumors were recruited. Tumors were reviewed by expert pathologists. Patients were prospectively followed-up (median, 62.6 months for disease-free patients) through review of hospital records and telephone interviews. The sequence of exons 7 and 10 of FGFR3 was analyzed by polymerase chain reaction and direct sequencing. We assessed the association of mutations with stage and grade. The predictive value of mutations for recurrence, progression, and mortality were assessed using Kaplan-Meier and Cox multivariable models. Results Mutations were more common among low malignant potential neoplasms (LMPN; 77%) and TaG1/TaG2 tumors (61%/58%) than among TaG3 tumors (34%) and T1G3 tumors (17%). The S249C, Y3...

Research paper thumbnail of Clinical Interest of LMO2 Testing for the Diagnosis of Aggressive Large B-Cell Lymphomas

Cancers

MYC rearrangements usually confer aggressive biological behavior to large B-cell lymphomas. In th... more MYC rearrangements usually confer aggressive biological behavior to large B-cell lymphomas. In this study, we aimed to evaluate the relevance of LMO2 detection to the clinical approach to these tumors. First, the ability of LMO2 loss of expression to recognize the presence of MYC rearrangements was evaluated. A series of 365 samples obtained from 351 patients, including 28 Burkitt lymphoma, 230 diffuse large B-cell lymphoma, 30 high-grade B-cell lymphoma with MYC and BCL2/BCL6 rearrangements, eight high-grade B-cell lymphoma-NOS, 43 transformed diffuse large B-cell lymphoma, and 26 high-grade follicular lymphomas was analyzed. Among the CD10-positive tumors prospectively analyzed in whole tissue sections, LMO2 negative expression obtained values of 88% sensitivity, 94% specificity, and 93% accuracy, proving the utility of LMO2 to screen MYC rearrangements. In addition, survival analyses were performed in a series of 155 patients. As per univariate analyses, the prognosis relevance o...

Research paper thumbnail of Complementary value of electron microscopy and immunohistochemistry in the diagnosis of non-small cell lung cancer: A potential role for electron microscopy in the era of targeted therapy

Ultrastructural Pathology, 2019

With the identification of therapeutic targets for lung adenocarcinoma, it has become mandatory t... more With the identification of therapeutic targets for lung adenocarcinoma, it has become mandatory to distinguish it from other entities. Some cases remain classified as non-small cell lung carcinoma, not otherwise specified (NSCLC-NOS) with immunohistochemistry. Electron microscopy (EM) can be useful, allowing the identification of glandular differentiation. The aim of this study was to determine the complementary value of immunohistochemistry and EM. Forty-eight NSCLC-NOS cases were selected (PSMAR-Biobank, Barcelona, Spain). Immunohistochemistry (TTF-1, p40) was performed. Tissue was retrieved from paraffin blocks. Results were compared to the final diagnosis, derived from combination of light microscopy, immunohistochemistry, EM, molecular studies and resection specimen. Immunohistochemistry concurred with final diagnosis in 36 cases (75%, Kappa = 0.517). EM agreed with final diagnosis in 35 (72.9%, Kappa = 0.471). Immunohistochemistry had a sensitivity = 73%, specificity = 100%, positive predictive value (PPV) = 100% and negative predictive value (NPV) = 52.4% for adenocarcinoma. All adenocarcinoma cases not solved by immunohistochemistry (n = 10) were classified by EM, and vice versa. Data from EM were identical to those of immunohistochemistry: sensitivity = 73%, specificity = 100%, PPV = 100% and NPV = 52.4%. Combining both techniques, 47 cases were coincident with final diagnosis (97.9%, Kappa = 0.943). EM can provide valuable information in subtyping NSCLC-NOS, being particularly useful when immunohistochemistry is inconclusive. EM could be considered as a complementary tool for decision-making in NSCLC-NOS.

Research paper thumbnail of ERG overexpression plus SLC45A3 (prostein) and PTEN expression loss: Strong association of the triple hit phenotype with an aggressive pathway of prostate cancer progression

Oncotarget, Sep 26, 2017

TMPRSS2 and SLC45A3 rearrangements may coexist in the same tumor. ERG rearrangements and PTEN los... more TMPRSS2 and SLC45A3 rearrangements may coexist in the same tumor. ERG rearrangements and PTEN loss are concomitant events in prostate cancer (PrCa), and can cooperate in progression. We have reported that mRNA expression of TMPRSS2-ERG and SLC45A3-ERG rearrangements plus PTEN loss define an aggressive tumor subset. The aim of this study has been to validate these results by immunohistochemistry in a large cohort of tumors. ERG, SLC45A3 and PTEN immunostaining and their association with pathological features and PSA progression-free survival were analyzed in 220 PrCa (PSMAR-Biobank, Barcelona, Spain). ERG protein expression was found in 46.8% and SLC45A3 and PTEN loss in 30% and 34% tumors, respectively. Single ERG positive immunostaining was associated with GS = 6 tumors (p = 0.016), double ERG+/PTEN loss with GS = 7 (p = 0.008) and Grade Group 2 (GG) or GG3 cases (p = 0.042), ERG+/SLC45A3 loss/PTEN loss ("triple hit") with GS ≥ 8 (p < 0.0001) and GG4 or GG5 tumors (p =...

Research paper thumbnail of FOXO1 down-regulation is associated with worse outcome in bladder cancer and adds significant prognostic information to p53 overexpression

Research paper thumbnail of 803 PI3K signalling pathway is activated by PIK3CA gain and overexpression in prostate tumours, but PI3KCA, BRAF, KRAS AND AKT1 mutations are an infrequent event

European Journal of Cancer Supplements, 2010

tumourigenesis in this subgroup remain poorly described. We have analysed copy number aberrations... more tumourigenesis in this subgroup remain poorly described. We have analysed copy number aberrations and mutation profile of known target genes in tumours from early and late onset CRC patients. Material and Methods: High resolution array CGH analysis (385 000 features), was performed in 23 patients with CRC diagnosis at young age (range: 28−53 years, median: 44 years), and 17 patients with CRC diagnosis at old age (range: 69−78 years, median: 79 years). Furthermore, mutation profiles were analysed in a larger series of carcinomas stratified according to microsatellite instability analysis (MSI) status. These patients included 45 young-at-onset (range; 27−50 years, median; 43 years), and 69 old-atonset (n = 69, range; 71−93 years, median; 81 years). A panel of five genes, TP53, KRAS, BRAF, PTEN and PIK3CA, were investigated for mutations by sequencing. Results: The overall genome copy number profiles were similar between carcinomas from patients in the two age groups. However, some chromosomal stretches were found to have statistically significant (p < 0.05) more aberrations in the young patients compared to the old-at-onset group (not visa versa); DNA sequences within 2q, 10q, 19q, were more often gained, and sequences within 1p, 1q, 2q, 4p, 4q 10p and 19p, were more frequently lost. KRAS and PTEN mutations were distributed equally between the patient groups, whereas the mutation frequencies of TP53 and PIK3CA differed between the groups. BRAF mutations were not significantly correlated with MSI in the young-atonset group. Conclusions: We have identified genomic and gene specific differences in colorectal carcinomas related to time of disease onset. The somatic genomic changes that occur preferentially in tumours from young patients pinpoint potential genetic risk loci that will be further examined.

Research paper thumbnail of Association of ERG and TMPRSS2-ERG with grade, stage, and prognosis of prostate cancer is dependent on their expression levels

The Prostate, 2015

BACKGROUND. There is controversy in the literature on the role of the fusion TMPRSS2-ERG in the p... more BACKGROUND. There is controversy in the literature on the role of the fusion TMPRSS2-ERG in the pathogenesis and progression of prostate cancer. The quantitative differences in TMPRSS2-ERG fusion expression have received very limited attention in the literature. METHODS. We have quantitatively analyzed the mRNA levels of TMPRSS2-ERG, ERG, PTEN, and AR (n ¼ 83), as well as ERG immunostaining (n ¼ 78) in a series of prostate tumors. RESULTS. Among the TMPRSS2-ERG cases (n ¼ 57), high fusion levels were associated with GS !8 (P ¼ 0.025). ERG mRNA overexpression was associated with GS !8 (P ¼ 0.047), and with stage T3-T4 tumors (P ¼ 0.032). Among the ERG overexpressing cases (n ¼ 54), higher expression levels were found in 92.3% of GS !8 tumors (P ¼ 0.02). ERG immunostaining, regardless of staining intensity, was also associated with high stage (P ¼ 0.05). There was a statistical association between ERG immunostaining and PSA progression-free survival (Log Rank test, P ¼ 0.048). Decreased PTEN expression was associated with TMPRSS2-ERG (P ¼ 0.01), ERG mRNA overexpression (P ¼ 0.003) and ERG immunostaining (P ¼ 0.007). Furthermore, decreased PTEN expression, alone (P ¼ 0.041) and also combined with TMPRSS2-ERG (P ¼ 0.04) or with ERG overexpression (P ¼ 0.04) was associated with GS !7 tumors. CONCLUSIONS. Although more studies are needed to further clarify their role, our findings emphasize that the expression levels of the TMPRSS2-ERG fusion and ERG mRNA, rather than their mere presence, are related to a more aggressive phenotype, have an effect on prognosis and could be molecular markers of progression for prostate cancer. Furthermore, ERG immunohistochemistry could be also a potentially useful prognostic factor.

Research paper thumbnail of PI3K signaling pathway is activated by PIK3CA mRNA overexpression and copy gain in prostate tumors, but PIK3CA, BRAF, KRAS and AKT1 mutations are infrequent events

Modern Pathology, 2010

The phosphatidylinositol 3-kinase (PI3K)-AKT and RAS-MAPK pathways are deregulated in a wide rang... more The phosphatidylinositol 3-kinase (PI3K)-AKT and RAS-MAPK pathways are deregulated in a wide range of human cancers by gain or loss of function in several of their components. Our purpose has been to identify genetic alterations in members of these pathways in prostate cancer. A total of 102 prostate tumors, 79 from prostate cancer alone (group G1) and 23 from bladder and prostate cancer patients (G2), are the subject of this study. In 20 of these 23, the bladder tumors were also analyzed. PIK3CA, KRAS, BRAF and AKT1 mutations were analyzed by direct sequencing, and BRAF also by pyrosequencing. PIK3CA quantitative mRNA expression and fluorescence in situ hybridization (FISH) gains were tested in 25 and 32 prostate tumors from both groups (G1 and G2), respectively. Immunohistochemistry for pAKT was performed in 55 prostate tumors. Of 25 prostate tumors, 10 (40%) had PIK3CA mRNA overexpression that was statistically associated with Gleason score Z7 (P ¼ 0.018). PIK3CA copy gain was detected in 9 of 32 (28%) prostate tumors. Of 20 bladder tumors, 3 (15%) displayed mutations in PIK3CA, KRAS and AKT1, the corresponding prostate tumors being wt. We also detected a previously not reported PIK3CA polymorphism (IVS9 þ 91) in two prostate tumors. In all, 56% of prostate tumors overexpressed pAKT. There is a statistical association (Po0.0001) of strong pAKT immunostaining with high Gleason score, and with PIK3CA alterations (mRNA overexpression and/or FISH gains). PIK3CA gene is deregulated by mRNA overexpression and DNA gain in B40 and 28% of prostate tumors, respectively. High-grade prostate tumors are associated with PIK3CA mRNA overexpression, but not with FISH status. PIK3CA, BRAF, KRAS and AKT1 mutations are very infrequent events in prostate tumors. However, PI3K signaling pathway is activated by PIK3CA FISH gain and/or mRNA overexpression, leading to an increased pAKT protein expression.

Research paper thumbnail of PIK3CA Mutations Are an Early Genetic Alteration Associated with FGFR3 Mutations in Superficial Papillary Bladder Tumors

Cancer Research, 2006

Bladder tumors constitute a very heterogeneous disease. Superficial tumors are characterized by a... more Bladder tumors constitute a very heterogeneous disease. Superficial tumors are characterized by a high prevalence of FGFR3 mutations and chromosome 9 alterations. High-grade and muscle-invasive tumors are characterized by Tp53 mutations and aneuploidy. We have analyzed the sequence of exons 9 and 20 of PIK3CA in a panel of bladder tumors covering the whole spectrum of the disease. DNA from formalin-fixed, paraffin-embedded tumor sections was amplified by PCR and products were sequenced. In an unselected panel of tumors representative of the disease, the PIK3CA mutation prevalence was 13% (11 of 87). Mutations occurred mainly at the previously identified hotspots (codons 542, 545, 1007, and 1047). The distribution according to stage was as follows: papillary urothelial neoplasms of uncertain malignant potential (PUNLMP; 11 of 43, 25.6%), Ta (9 of 57, 16%), T1 (2 of 10, 20%), and muscle-invasive tumors (0 of 20, 0%; P = 0.019). Mutations were associated with low-grade tumors: grade 1 ...

Research paper thumbnail of Abstract 1168: Urothelial cell carcinomas harbor very frequent mutations in FGFR3-PI3K-AKT pathway and highest prevalence of mutations is related to low-grade tumors

Cancer Research, 2010

More than 90% of bladder cancers are urothelial cell carcinomas. Superficial/papillary and invasi... more More than 90% of bladder cancers are urothelial cell carcinomas. Superficial/papillary and invasive tumors progress via different molecular pathways. FGFR3 and PI3KCA mutations are more commonly found in superficial/papillary tumors. The PI3K-AKT pathway emerges as an alternative pathogenic mechanism linked to bladder cancer. We have analyzed the prevalence of mutations in FGFR3, PI3KCA, KRAS, BRAF and AKT1 genes in a set of 61 bladder tumors classified as TaG1 (n=10), TaG2 (n=19), TaG3 (n=8), T1G2 (n= 3), T1G3 (n= 11), T2G2 (n=1) and T2G3 (n=10). We focused on analyzing the hot spot mutation codons and surrounding regions. Mutation analysis of DNA from paraffin-embedded tumors was performed by direct sequencing of PCR products. Thirty-six (60%) tumors presented mutations, 22 cases with only one mutated gene, whereas 14 of 61 cases had mutations in two genes. The most frequently mutated gene was FGFR3 in 25 tumors, followed by PI3KCA in 16 tumors. The distribution and combinations o...

Research paper thumbnail of CXCR4 mRNA overexpression in high grade prostate tumors: lack of association with TMPRSS2-ERG rearrangement

The TMPRSS2-ERG fusion has been reported in 42 to 78% of prostate tumors. More than 90% of ERG-ov... more The TMPRSS2-ERG fusion has been reported in 42 to 78% of prostate tumors. More than 90% of ERG-overexpressing tumors harbor the fusion. The relationship between the TMPRSS2-ERG fusion and prognosis is controversial. Different studies have suggested an association between CXCR4 and ERG overexpression resulting from the TMPRSS2-ERG rearrangement. The aim of this study was to investigate the relationship between CXCR4 expression, TMPRSS2-ERG fusion and Gleason grade in prostate cancer. TMPRSS2-ERG rearrangement was investigated by FISH (n = 44), ERG protein by IHC (n = 84), and CXCR4 by quantitative RT-PCR (n = 44). TMPRSS2-ERG rearrangement and ERG protein expression were present in almost 50% of the cases, without statistical differences between the different Gleason score groups. There was a very high concordance between FISH and IHC techniques (Kappa Index = 0.954). Seventy percent of Gleason 8 prostate tumors overexpressed CXCR4 mRNA, and the difference in CXCR4 expression with Gleason < 8 cases was statistically significant (p = 0.009). There was no association between ERG protein and CXCR4 mRNA expression. In conclusion, our results reveal for the first time that CXCR4 overexpression is associated with high Gleason score prostate tumors, but that it is independent of the TMPRSS2-ERG rearrangement.

Research paper thumbnail of SPOP and CHD1 alterations in prostate cancer: Relationship with PTEN loss, tumor grade, perineural infiltration, and PSA recurrence

Research paper thumbnail of Immunohistochemical expression of mismatch repair proteins (MSH2, MSH6, MLH1, and PMS2) in prostate cancer: correlation with grade groups (WHO 2016) and ERG and PTEN status

Research paper thumbnail of SPOP and FOXA1 mutations are associated with PSA recurrence in ERG wt tumors, and SPOP downregulation with ERG ‐rearranged prostate cancer

Research paper thumbnail of Strong cytoplasmic ETV1 expression has a negative impact on prostate cancer outcome

Research paper thumbnail of Hernandez, S. et al. CDC25A and the splicing variant CDC25B2, but not CDC25B1, -B3 or -C, are over-expressed in aggressive human non-Hodgkin's lymphomas. Int. J. Cancer 89, 148-52

International Journal of Cancer

ABSTRACT

Research paper thumbnail of 816 FGFR3 mutations in prostate cancer and other tumours

European Journal of Cancer Supplements, 2010

Background: We previously reported FGFR3 mutations in prostate carcinoma (PCa). FGFR3 mutations w... more Background: We previously reported FGFR3 mutations in prostate carcinoma (PCa). FGFR3 mutations were associated with low-grade PCa, and also with PCa found in patients with concurrent bladder cancer or skin tumours. The aim of this work has been to further investigate the relationship between FGFR3 alterations in PCa and the presence of concurrent tumours. Material and Methods: 41 cases with PCa and other associated tumour types were studied. The PCa series consisted of: 22 incidental (cystoprostatectomy) tumours and 19 clinically significant (biopsy or prostatectomy) cases. Twentythree PCa were Gleason grade 6, 11 were Gleason grade 7 and 7 were Gleason grade 8. In each case, we studied the PCa and the concurrent tumour (prostate and bladder cancer, n = 32; prostate and skin tumour, n = 6; prostate and colon cancer, n = 2; prostate and lung cancer, n = 1). FGFR3 exons 7 and 10 were analysed by PCR and direct sequencing. Results: Eight of 41 (19.5%) PCa presented a mutation in FGFR3. From these, 6 were Gleason grade 6, and 2 were Gleason grade 7. Four of 32 (13%) patients with PCa and bladder cancer harboured a FGFR3 mutation in the PCa, and 5 other cases (16%) in the bladder tumour. In the PCa-skin tumour group, 3 of 6 (50%) PCa presented a FGFR3 mutation, and other 2 different cases (33%) in the skin tumour. One case harboured FGFR3 mutations in both tumours, but in different codons. Finally, one case with PCa and colon cancer also had a FGFR3 mutation in PCa. Conclusions: FGFR3 mutations in PCa are associated with an increased frequency of concurrent tumours in other organs, mainly skin and bladder. The lack of coincidence in the presence of FGFR3 mutations in both the PCa and the associated tumours suggests that they evolve through different pathways.

Research paper thumbnail of TRK-fused gene (TFG) is a new partner of ALK in anaplastic large cell lymphoma producing two structurally different TFG-ALK translocations

Blood, 1999

Anaplastic large cell lymphoma (ALCL) is associated with the t(2;5)(p23;q35), which generates the... more Anaplastic large cell lymphoma (ALCL) is associated with the t(2;5)(p23;q35), which generates the NPM-ALK fusion gene encoding an 80-kD protein. Several studies have suggested that genes other than NPM may be fused to the ALK gene. Here we have identified TRK-fused gene (TFG) as a new ALK partner in 2 ALCL, 1 of which exhibited a t(2;3)(p23;q21). In these cases, TFG was involved in 2 different fusion genes, TFG-ALK(S) and TFG-ALK(L), coding respectively 85-kD and 97-kD chimeric proteins. The ALK breakpoint in these translocations was the same as in the classic t(2;5) translocation. These 2 proteins were both active in an in vitro tyrosine kinase assay showing that the new cloned cDNA sequences are translated into chimeric proteins with functional activity. These findings indicate that TFG can provide an alternative to NPM as a fusion partner responsible for activation of the ALK and the pathogenesis of ALCL.

Research paper thumbnail of Mutations in FGFR3 and PIK3CA, singly or combined with RAS and AKT1, are associated with AKT but not with MAPK pathway activation in urothelial bladder cancer

Human Pathology, 2012

Different members of the phosphoinositide 3 kinase--serine threonine protein kinase (PI3K-AKT) pa... more Different members of the phosphoinositide 3 kinase--serine threonine protein kinase (PI3K-AKT) pathway are altered in bladder cancer. Fibroblast growth factor receptor 3 (FGFR3) mutations characterize the low-grade tumors, and RAS genes are mutated in approximately 13% of all bladder tumors. Interestingly, a percentage of bladder tumors have alterations in more than 1 PI3K-AKT or rat sarcoma viral oncogene homolog-RAF mitogen activated protein kinase (RAS-MAPK) pathway gene or their upstream regulators, but some combinations are mutually exclusive. We analyzed mutations in FGFR3, phosphoinositide 3 kinase catalytic alpha polypeptide (PIK3CA), v-akt murine thymoma viral oncogene homolog 1 (AKT1), v-Ki-ras2 Kirsten rat sarcoma viral oncogene homolog (KRAS), v-Ha-ras Harvey rat sarcoma viral oncogene homolog (HRAS), and v-raf murine sarcoma viral oncogene homolog B1 (BRAF) in 88 urothelial cell carcinomas and the immunohistochemical expression of phospho-v-akt murine thymoma viral oncogene homolog (AKT) and mitogen-activated protein kinase 1 and 2 (pERK1/2) in 80 and 77 urothelial cell carcinomas, respectively. Approximately 43% and 20.5% of tumors presented 1 and 2 mutated genes, respectively. FGFR3 mutations were more frequent alone, whereas PIK3CA mutations were associated with another mutated gene (FGFR3 and KRAS). Overall, mutated FGFR3 (FGFR3(mut)) and mutated FGFR3 (FGFR3(mut))-mutated PIK3CA (PIK3CA(mut)) genotypes were associated with low-grade bladder tumors and mutated PIK3CA (PIK3CA(mut))-mutated KRAS (KRAS(mut)) and mutated AKT1 (AKT1(mut)) were only present in high-grade tumors. There are no mutated FGFR3 (FGFR3(mut))-mutated RAS (RAS(mut)) nor mutated PIK3CA (PIK3CA(mut))-mutated AKT1 (AKT1(mut)) combinations. Fifty percent and 56% of tumors showed high levels of pAKT and pERK1/2, respectively. High levels of pAKT were associated with total mutations, FGFR3(mut), and PIK3CA(mut) tumors but not with tumor grade or stage. Wild-type tumors presented significantly higher pERK1/2 expression. Mutations in FGFR3 and FGFR3-PIK3CA but not single PIK3CA mutations characterize low-grade bladder tumors. Single FGFR3 or PIK3CA mutations and the different mutation combinations FGFR3-PIK3CA/AKT1 and PIK3CA-RAS can activate the AKT but not the MAPK pathway. Other genes different from FGFR3 may be related with the pERK activation in bladder tumors.

Research paper thumbnail of Manual Versus Laser Micro-dissection in Molecular Biology

Ultrastructural Pathology, 2006

Molecular studies on whole samples of fresh or frozen tissue do not take into account the heterog... more Molecular studies on whole samples of fresh or frozen tissue do not take into account the heterogeneity of these tissues. In addition to normal cells, precursor lesions and different progression stages may be mixed within a given sample. Usually, the dominant cell population will determine the results and may sometimes mask biologically relevant abnormalities. To obtain more specific information and knowledge on changes within different cell compartments, many techniques have been developed that combine morphological observation and selection with different strategies for specific cell dissection. In this review, the most important micro-dissection methods are put into perspective, and some requirements and limitations are discussed with regard to sample fixation, staining, dissection and molecular analysis.