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The majority of metastatic renal cell carcinoma (RCC) patients are treated with tyrosine kinase inhibitors (TKI) in first-line treatment; however, a fraction are refractory to these antiangiogenic drugs. MicroRNAs (miRNAs) are regulatory molecules proven to be accurate biomarkers in cancer. Here, we identified miRNAs predictive of progressive disease under TKI treatment through deep sequencing of 74 metastatic clear cell RCC cases uniformly treated with these drugs. Twenty-nine miRNAs were differentially expressed in the tumors of patients who progressed under TKI therapy (P values from 6 × 10-9 to 3 × 10-3). Among 6 miRNAs selected for validation in an independent series, the most relevant associations corresponded to miR-1307-3p, miR-155-5p, and miR-221-3p (P = 4.6 × 10-3, 6.5 × 10-3, and 3.4 × 10-2, respectively). Furthermore, a 2 miRNA-based classifier discriminated individuals with progressive disease upon TKI treatment (AUC = 0.75, 95% CI, 0.64-0.85; P = 1.3 × 10-4) with better predictive value than clinicopathological risk factors commonly used. We also identified miRNAs significantly associated with progression-free survival and overall survival (P = 6.8 × 10-8 and 7.8 × 10-7 for top hits, respectively), and 7 overlapped with early progressive disease. In conclusion, this is the first miRNome comprehensive study, to our knowledge, that demonstrates a predictive value of miRNAs for TKI response and provides a new set of relevant markers that can help rationalize metastatic RCC treatment.

KISS1 is a metastasis suppressor lost in several solid malignancies. We evaluated the clinical relevance of KiSS-1 methylation and its protein expression in colorectal cancer. The epigenetic silencing of KiSS-1 by hypermethylation was tested in colon cancer cells (n = 5) before and after azacytidine treatment. KiSS-1 methylation was evaluated by methylation-specific PCR in colorectal cancer cells, and normal, benign, and tumor tissues (n = 352) were grouped in a training set (n = 62) and two independent validation cohorts (n = 100 and n = 190). KiSS-1 protein expression was analyzed by immunohistochemistry on tissue arrays. KiSS-1 hypermethylation correlated with transcript and protein expression loss, being increased in vitro by azacytidine. Methylation rates were 53.1, 70.0, and 80.0 % in the training and validation sets, respectively. In the training set, KiSS-1 methylation rendered a diagnostic accuracy of 72.7 % (p = 0.002). Combination of KiSS-1 methylation and serum CEA (p = 0.001) increased the prognostic utility of CEA alone (p = 0.022). In the first validation set, KiSS-1 methylation correlated with tumor grade (p = 0.011), predicted recurrence (p = 0.009), metastasis (p = 0.004), disease-free (p = 0.034), and overall survival (p = 0.015). In the second validation cohort, KiSS-1 methylation predicted disease-specific survival (p = 0.030). In the training set, cytoplasmic KiSS-1 expression was significantly higher in nonneoplastic biopsies as compared to colorectal tumors (p

The banks of tumours exist since the pathological anatomy and cytology laboratories preserve tissue samples, in particular by paraffin inclusion. Nevertheless, the current biotechnological projections changed our needs to access to tissues and other biological samples.

Background Previous studies suggest that expression of hypoxia markers may be associated with response to antiangiogenic drugs. Thus, we aimed to identify predictors of sunitinib outcome in clear-cell renal cell carcinoma (ccRCC). Patients and methods The expression of eight key proteins related to hypoxia (CAIX, HIF1A, HIF2A, VEGFA, VEGFR1, VEGFR2, VEGFR3 and PDGFRB) and P-glycoprotein were assessed by immunohistochemistry in 67 primary ccRCC samples from prospectively recruited patients treated with first-line sunitinib. The proteins expression, VHL inactivation and EGLN3 mRNA content were compared with the patients' response to sunitinib. Results High expression of HIF2A and PDGFRB was associated with better sunitinib RECIST objective response (P = 0.024 and P = 0.026; respectively) and increased VEGFR3 expression was associated with longer progression-free survival (P = 0.012). VEGFR3 overexpression showed a negative correlation with VEGFR3 polymorphism rs307826 (P = 0.002), a sunitinib resistance predictor. With respect to overall survival (OS), high VEGFA was associated with short (P = 0.009) and HIF2A with long (P = 0.048) survival times. High EGLN3 mRNA content was associated with shorter OS (P = 0.023). Conclusions We found an association between several proteins involved in hypoxia and sunitinib efficacy. In addition, low VEGFR3 expression was associated with worse outcome and with VEGFR3 rs307826 variant allele, reinforcing VEGFR3 as a marker of sunitinib resistance.

Aberrant overexpression of cyclooxygenase-2 (COX2) is observed in urothelial carcinoma of the bladder (UCB). Studies evaluating COX2 as a prognostic marker in UCB report contradictory results. We determined the prognostic potential of COX2 expression in UCB and quantitatively summarize the results with those of the literature through a meta-analysis. Newly diagnosed UCB patients recruited between 1998–2001 in 18 Spanish hospitals were prospectively included in the study and followed-up (median, 70.7 months). Diagnostic slides were reviewed and uniformly classified by expert pathologists. Clinical data was retrieved from hospital charts. Tissue microarrays containing non-muscle invasive (n = 557) and muscle invasive (n = 216) tumours were analyzed by immunohistochemistry using quantitative image analysis. Expression was evaluated in Cox regression models to assess the risk of recurrence, progression and disease-specific mortality. Meta-hazard ratios were estimated using our results and those from 11 additional evaluable studies. COX2 expression was observed in 38% (211/557) of non-muscle invasive and 63% (137/216) of muscle invasive tumors. Expression was associated with advanced pathological stage and grade (p

BACKGROUND: Triple-negative breast cancer (TNBC) is a breast cancer subtype that is not defined by targetable molecular markers. Since all the aberrations that exist and ultimately contribute to the tumor phenotype converge, from a functional point of view, in the final status of the phosphorylation of the proteome in a given moment, we sought to interrogate the phosphoproteome with two aims: 1) establish a taxonomy of TNBC based on measurable markers that predict clinical course; 2) reduce the phosphoproteome that characterizes the bad- from good-prognosis cases to its targetable, driving kinases, in order to define rational therapeutic approaches in TNBC. We chose mass spectrometry as a discovery approach, because of its coverage, sensitivity, dynamic range and specificity. Here we report the part of phosphoproteomic analysis used for establishing a new taxonomy of TNBC. METHODS: We performed quantitative phosphoproteomics from a discovery set of 34 frozen tumor samples divided in two sets paired by classic prognosis factors: 1, with 13 patients relapsed in less than 3 years; 2, 21 patients, no relapse in 10 years follow up. Raw data were processed with Maxquant and cases clustered using a supervised hierarchical approach. Kinases (predicted with linear domain consensus analysis tools using a PHOSIDA) and phospho-sites discriminating the set 1 from 2 were validated by immunohistochemistry in 113 consecutive TNBC cases, spotted in TMAs, with 14 years follow up. RESULTS: Overall, 15000 unique phosphopeptides were identified. Supervised clustering of subset 1 vs. 2 showed that 161 and 541 peptides were significantly more phosphorylated in the subset 1 and 2, respectively (FDR Kinase validation was performed directing an antibody against the active kinase form, if available; if not, against the total levels. The staining of kinases and the candidate phosphosites was analyzed with an Ariol, scored with a continue value from 0 to 3, and divided in quartiles for each staining probe. So far, 12 probes have been quantified. The prognosis of patients with upper quartile staining vs. the remainder was analyzed with the log-rank test and cox proportionate hazards model for each probe. Five of them showed statistically significant association with relapse (pSTAT3Tyr705, 6.1 vs. 10.1 years, p = 0.024; CYCB1, 7 vs. 10.3 years, p = 0.027; CDK6, 7 vs. 10.3 years, p = 0.013; pp70S6KThr389, 7.2 vs. 10 years, p = 0.042 and PRKCEpsilon, 7.1 vs. 10.2 years, p = 0.021). A combined variable integrating upper-quartile staining from either of those 5 probes occurring in 67.4% patients, identified almost all patients that experienced relapse: 48.3% of them relapsed (median time 7.6 years) vs. the reminder 32.6% of which only 13.8% relapsed (median time 11.9 years), log rank p = 0.001 INTERPRETATION: High-throughput mass-spectrometry is a powerful tool for generating a disease taxonomy, and can be translated to routine techniques like IHC. The signature constituted by pSTAT3Tyr705, CYCB1, CDK6, pp70S6K and PRKCEpsilon constitutes the most parsimonious signature to date able to detect all the early TNBC patients that relapse in the long term. Citation Format: Ivana Zagorac, Tamara Mondejar Tevar, Jesús Sánchez Ruíz, Albert Heck, Maarten Altelaar, Renske Penning, Harm Post, Gonzalo Gómez López, David Pisano, Javier Muñoz Peralta, Javier Muñoz Peralta, Manuel Morente, Luis Manso, Miguel Quintela-Fandiño. A phosphoproteomic portrait of triple - negative breast cancer: functional taxonomy. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2015 Nov 5-9; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2015;14(12 Suppl 2):Abstract nr B9.

The International Prognostic Score (IPS) is the most widely used system to date for identifying risk groups for the outcome of patients with advanced Hodgkin lymphoma, although important limitations have been recognized. We analyzed the value of the IPS in a series of 311 patients with advanced classical Hodgkin lymphoma (cHL) (Ann Arbor stage III, IV or stage II with B symptoms and/or bulky masses) treated with first-line chemotherapy including adriamycin (adriamycin, bleomycin, vinblastine, dacarbazine [ABVD] or equivalent variants). In univariate and multivariate analyses, stage IV disease and age ≥ 45 years were the only factors with independent predictive significance for overall survival (OS) (p = 0.002 and p < 0.001, respectively). Stage IV was still significant for freedom from progression (FFP) (p = 0.001) and age ≥ 45 years was borderline significant (p = 0.058). IPS separates prognostic groups, as in the original publication, but this is mainly due to the high statistical significance of stage IV and age ≥ 45 years. Moreover, the combination of these two factors enables a simpler system to be constructed that separates groups with different FFP and OS. In conclusion, in our series, stage IV and age ≥ 45 years are the key prognostic factors for the outcome of advanced cHL.

The banks of tumours exist since the pathological anatomy and cytology laboratories preserve tissue samples, in particular by paraffin inclusion. Nevertheless, the current biotechnological projections changed our needs to access to tissues and other biological samples.

To assess the internal consistency, discriminative capacity and factorial composition of the Professional Quality of Life Questionnaire (QPL-35) in a population of primary care professionals.We performed a cross-sectional analytical study in a primary care area in Madrid from 2001 to 2003. Random sampling of 450 healthcare professionals was performed on 2 occasions. The sample was stratified into 3 groups: group I (clinicians, pharmacologists, psychologists), group II (nurses, midwives, physiotherapists, social workers) and group III (administrative staff, porters, auxiliary nurses). The self-administered questionnaire QPL-35 was sent in January 2001 and January 2003 and on each occasion the questionnaire was sent again 1 month later. The percentages of total responses and responses per item were studied. We also studied the distribution of each answer by examining the floor effect and ceiling effect, as well as the factorial composition based on a previous validation study.Five hundred sixty-three questionnaires (62.6%) were returned. All the questions had a response rate of more than 96%. At least one unanswered question was found in 22.0% of the questionnaires, and at least 2 were unanswered in 7.1%. The distribution of the answers did not fit normal distribution in any of the cases. The floor effect was present in questions related to management support and the ceiling effect was found in those related to motivation. The factorial analysis found 3 factors that explained 39.6% of the variance in the total number of questions. These factors were very similar to those of the previous validation study: management support, perception of workload and intrinsic motivation explained 17.0%, 13.2% and 9.4% of the variance, respectively. Internal consistency was high for each factor (Cronbach's alpha0.7) and for the total score (Cronbach's alpha = 0.81).The metric properties of the QPL-35 are maintained in different environments. This questionnaire can be recommended as a tool to measure and compare quality of professional life in primary care.

Introduction Sunitinib is a standard treatment for kidney cancer, however in some patients response is lacking. Molecular predictors of outcome could deeply impact patient care. Methods We conducted an observational, prospective study in 15 institutions members of the Spanish Oncology Genitourinary Group (SOGUG). Inclusion criteria were patients with confirmed advanced clear cell renal cell carcinoma with no prior treatment, planned to receive sunitinib according to local practice. From paraffin embedded tumor samples a Tissue MicroArray (TMA) was constructed and evaluated independently by two pathologists, also blinded to clinical data. Expression of Carbonic Anhydrase IX, P-glicoprotein, Vascular Endothelial Growth Factor (VEGF), VEGF-Receptor 1,2 and 3 (VEGFR1,2 and 3), Platelet-Derived Growth Factor Receptor (PDGFR-Beta), Hypoxia-Inducible Factor 2 alpha (HIF2α) and Von Hippel-Lindau protein (pVHL) was assessed. In addition the presence of VHL gene mutations was studied. Results 101 patients were recruited from 2007 to 2010. TMA was constructed from 69 tumor samples. In multivariable analysis, HIF2α (HR 0.17 (0.05-0.64) p = 0.0083) and PDGFRβ (HR 0.042 (0.003-0.70) p = 0.028) overexpression correlated with progressive disease (PD) as best response. A similar trend in PFS and OS was observed for HIF-alfa but did not reach statistical significance. Progression free survival (PFS) was longer in cases with high VEGFR3 expression (HR 0.42 (0.21-0.83) p = 0.013). Interestingly, low levels of this protein strongly correlated (r = -0.441 P = 0.0003) with the presence of a germline SNP (VEGFR3 rs307826) that has been previously identified as a sunitinib resistance predictor by our group. Finally overexpression of VEGF (HR 4.6 (1.5-13.6), p= 0.0063), and VEGFR1 (HR 6.2 (1.2-32.2) p = 0.031) were associated with poor overall survival (OS). VHL gene alterations could only be determined in 30 cases, with 20 (66%) presenting a pathological mutation. No association with outcome could be established. Conclusion Expression of some proteins involved in neoangiogenesis pathway could play a role in sunitinib sensitivity and resistance. A significant association between low expression of VEGFR3 and one SNP in such gene, both correlated with poor outcome, deserves further investigation. Disclosure All authors have declared no conflicts of interest.

An immunohistochemical and ultrastructural study of 14 cases of persistent and generalized lymphadenopathy (PGL), acquired immune deficiency syndrome (AIDS) related, revealed florid follicular hyperplasia, follicular dendritic cell (FDC) lysis, lymphoid follicle invaginations, increased presence of T8 cells in germinal center, immature sinus histiocytosis (monocytoid B-cells), and inversion of T4/T8 ratio in the paracortical area. Electron microscopic examination showed viral particles of morphologic characteristics consistent with human immunodeficiency virus (HIV) virions attached to the processes of FDC in three of the nine cases studied. Lesions of the germinal center dendritic cell network are the cardinal feature of PGL. This finding lends support to the idea of a viral aggression directed against FDC as the cause of disregulation of the B-cells.

We sought to identify genetic variants associated with disease relapse and failure to hormonal treatment in hormone-receptor positive breast cancer (HRPBC). We analyzed a series of HRPBC with distant relapse, by sequencing pairs (n = 11) of tumors (primary and metastases) at >800X. Comparative genomic hybridization was performed as well. Top hits, based on the frequency of alteration and severity of the changes, were tested in the TCGA series. Genes determining the most parsimonious prognostic signature were studied for their functional role in vitro, by performing cell growth assays in hormonal-deprivation conditions, a setting that mimics treatment with aromatase inhibitors. Severe alterations were recurrently found in 18 genes in the pairs. However, only MYC, DNAH5, CSFR1, EPHA7, ARID1B, and KMT2C preserved an independent prognosis impact and/or showed a significantly different incidence of alterations between relapsed and non-relapsed cases in the TCGA series. The signature composed of MYC, KMT2C, and EPHA7 best discriminated the clinical course, (overall survival 90,7 vs. 144,5 months; p = 0.0001). Having an alteration in any of the genes of the signature implied a hazard ratio of death of 3.25 (p