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371 Background: Inflammation plays a key role in the pathophysiology of many diseases, including cancer. Systemic inflammatory factors have been validated as indicators of ongoing systemic inflammation that could be predictive markers of poor prognosis for oncological outcomes. However, it is unknown the prognostic impact of systemic inflammation markers in patients with GEP-NETs treated with PRRT. Methods: We conducted an observational, retrospective, multicentric study of 40 patients with GEP-NET treated with PRRT belonging to GGNET (Galician Research Group on Neuroendocrine Tumors) network at Nuclear Medicine Department of Santiago de Compostela University Hospital (Spain). The systemic inflammatory markers were calculated as follows: NLR = neutrophil count/lymphocyte count, PLR = platelet count/lymphocyte count, MLR= monocyte count/lymphocyte count, ALB= albumin levels and dNLR = neutrophil count/ (leucocytes count – neutrophils count). For the calculation of the different ratios, baseline analysis and after the second dose were used. The cut-off values were determined as the median of each values, correlating them with progression-free survival (PFS). Results: Data from 40 patients (pts) treated between 2016 and 2020 were recorded. Median age was 63.5 years (range 41-85) and 55% were male. Baseline ECOG PS 0/1/2 was 15 (37.5%)/16 (40%)/9 (22.5%). Tumor location was intestinal 26 pts (65%), pancreas 11 pts (27.5%) and unknown origin 3 pts (7.5%). 15 pts (37.5%) were functioning. Tumor grade G1/G2/G3 were 17 pts (42.5%)/ 20 pts (50%)/ 3 pts (7.5%), and Ki 67 20%/unknown were 11 pts (27.5%)/ 21 pts (52.5%)/ 3 pts (7.5%)/ 5 pts (12.5%), respectively. The most frequent site of metastasis was liver 32 pts (80%), lymph nodes 19 pts (47.5%), peritoneum 11 pts (27.5%) and bone 10 pts (25%). Surgery: 22 pts (55%) primary tumor surgery and 8 pts (20%) metastasectomy. Previous systemic treatments included somatostatin analogs (SSA) 40 pts (100%), everolimus 26 pts (65%) and sunitnib 11 pts (27.5%), others 7 pts (17.5%). The baseline cutoff-values for NLR was 2.61, for PLR 110.14, for MLR 0.31, for ALB 4.2. and for dNLR 1.71. The cutoff-values after the 2nd dose for NLR was 2.3, for PLR 2.15, for MLR 0.3, for ALB 4.2 and for dNLR 1.48. With a median follow up of 21 months, 14 pts (35%) had died. Median PFS was 27.2 m (95% CI 16.0-38.4m) and OS was not reached (NR). Pts with baseline higher NLR (>2.61 vs. 1.71 vs.

e13061 Background: Smoking is the leading cause of avoidable death in the world. Although controversial for years, current prospective studies suggest that smoking increases the risk of cancer in women with a deleterious BRCA1/2 mutation. Actually, we have very limited information about tobacco habit among BRCA1/2 carriers. Methods: We report an observational retrospective study of a consecutive sample of 198 BRCA1/2 carriers studied between April 2014 and March 2018 at the Family Cancer Clinic of the University Hospital of Coruña (NW-Spain). The main objective of the study is to know the prevalence of smoking in BRCA1/2 women, and to determine if carriers undergoing risk-reducing surgeries (RRS) abandon their tobacco habit as part of the primary prevention strategy. Results: The sample includes 136 patients affected by cancer (84% breast, 19% ovary) and 62 healthy women. During their follow-up [Median = 22, (1-43) months], 85 carriers underwent RRS (29% mammary, 54% prophylactic salpingo-oophorectomy, 16% both). At first visit, 46 were active smokers, 47 ex-smokers, 77 non-smokers and 28 did not specify their habit. The percentage of smokers and never smokers is similar between sick and healthy BRCA1/2 carriers, presenting a greater proportion of former smokers among those affected of cancer (30% vs. 23%), and of active smokers among healthy women (35% vs. 23%) [p > 0.5)]. Moreover, 23% of BRCA1/2 carriers are active smokers despite a previous diagnosis of cancer. RRS are not associated to a greater tobacco cessation, persisting in the habit 18% of the women that underwent any preventive surgery (p > 0.5). Conclusions: Smoking habit is high (55%) among BRCA1/2 carriers followed at our family cancer clinic. At least 23% of our BRCA1/2 positive women are active smokers despite a previous diagnosis of cancer.The performance of aggressive RRS is not associated with greater abandonment of tobacco in carriers, persisting in the smoking habit 18% of operated women.It is essential to improve anti-tobacco strategies in family cancer consultations if we want to be efficient in the management of high-risk cancer individuals.

12043Background: The determination of monitoring Circulating Tumor Cells (CTC) in patients (pts) with metastatic breast cancer (MBC) is a well-established prognostic marker of efficacy/tolerance. W...

468 Background: Detection of circulating tumor cells (CTC) may provide diagnostic and prognostic information in genitourinary tumors (GT). The aim of this work was identify aberrantly expressed miRNAs potentially useful for CTC detection in blood samples from patients with GT to assess their potential clinical significance, and to gain a greater understanding of the mechanisms driving tumor progression. Methods: We examined blood levels of 92 microRNAs in 113 metastatic patients: prostate (mP), renal cell carcinoma (mRCC), bladder tumors (mB) and healthy volunteers (HV) (N=18) using SYBR-green-based microRNA RT-qPCR arrays (Exiqon). Array design was made by literature review and bioinformatics approach using free databases. Relative Expression Software Tool (REST) and SPPS 21.0 were used to data processing and analysis. Results: microRNAs differential expressed between tumors and HV (only up-regulated miRNAs are shown in Table). Conclusions: microRNAs in the circulation are relatively stable, very accessible, low invasive and easily testable biomarkers. These results suggest that our bioinformatic approach is an useful high-throughput method to identified tumors-associated miRNAs. The selected miRNAs should be further evaluated for their potential as markers for CTC detection, prognosis and therapeutic outcome. Further studies of precision and sensitivity on these markers in this GT population will clarify its role as novel stable blood-based markers. [Table: see text]

451 Background: Renal cell carcinoma (RCC) is known to be resistant to chemotherapy. There is need for the identification of biomarkers capable to determine RCC prognosis factors and metastatic potential obtainable from non-invasive or minimally invasive techniques. Our aim was to derive predictive models which could predict more accurately than any one factor alone. Methods: To studythe cascade of events leading to the formation and progression of RCC, we assessed 29 markers by immunohistochemistry and qRT-PCR using tissue micro-array (TMA). Results: Multivariate logistic regression showed the best proteins combination for node status (NOTCH1 and GLUT5) and pelvis invasion (EGFR and DLL3). ROC curve analyses were made to analyse the accuracy of the best candidate proteins; it should be noted NOTCH1 and GLUT5 for node status prediction (AUC=0.833, 95% CI, 0.744-0.922; p

We describe a 5-month-old boy with complex congenital heart defects (dTGA, DORV, VSD, ASD, and PDA), minor facial and ear anomalies, deep palmar creases, multiple vertebral anomalies, agenesis of the corpus callosum, and mosaic tetrasomy 8p (47,XY,+i(8)(p10)[88%]/46,XY[12%] in blood with normal chromosomes in cultured skin fibroblasts. This infant represents the eleventh reported case of mosaic tetrasomy 8p since its first description by Kristofferson et al. [1988: Clin Genet 34:201-203]. The pattern of heart malformations and discordance of blood and fibroblast karyotypes make our case unique. Our report and review suggest that an important distinction between mosaic tetrasomy 8p and other chromosome 8 aneuploidies involves the increased incidence and complexity of congenital heart malformations.

Rearrangements in the vicinity of the centromere of chromosome 1 are over-represented in many types of human cancer and are a characteristic feature of a rare genetic disease called ICF (immunodeficiency, centromeric heterochromatin instability, and facial anomalies). Evidence is presented that implicates DNA hypomethylation in the formation of these pericentromeric chromosomal anomalies. The DNA methylation inhibitors 5-azadeoxycytidine and 5-azacytidine, but not other tested genotoxins, induced the preferential formation of pericentromeric rearrangements of chromosome 1 at a very high frequency in a pro-B-cell line (FLEB14) and at a lower frequency in a mature B-cell line (AHH-1). These abnormal chromosomes appear identical to the diagnostic chromosomal aberrations in the ICF syndrome. A major component of the pericentromeric DNA in chromosome 1, satellite 2, was shown to be hypomethylated in an ICF B-cell line, although DNA from this cell line did not display detectable overall hypomethylation. It is hypothesized that demethylation in certain DNA regions, including in pericentromeric satellite DNA, helps lead to pericentromeric chromosomal rearrangements in lymphocytes from ICF patients and in normal lymphoblastoid cells incubated in vitro with DNA demethylating agents.

5-Azacytidine (azaCR) causes genomic demethylation and decondensation of juxtacentromeric heterochromatin in chromosomes 1, 9, and 16. We determined the karyotypes of a pro-B cell line (FLEB14) treated with azaCR or its deoxynucleoside analog (azaCdR). About 80% of the induced rearrangements were in chromosome 1, and almost 90% of these involved its pericentromeric region. Multibranched figures with up to seven chromosome 1 arms, as well as whole-arm deletions of this chromosome, were the predominant anomalies, often with one normal homolog of chromosome 1 present. Isochromosomes 1 and fusions in the pericentromeric regions of chromosomes 1 and 16 or chromosomes 1 and 9 were also seen. The overlap of the spectrum of chromosomal rearrangements in azaCR- or azaCdR-treated FLEB14 cells and in mitogen-stimulated lymphocytes from patients with a rare genetic disease (ICF) associated with localized DNA hypomethylation supports the hypothesis that the DNA demethyiating activity of azaCR is essential for the induction of these pericentromeric rearrangements. These studies may help elucidate the overrepresentation of chromosome 1 pericentromeric rearrangements in many types of cancer cells.

Intrachromosomal rearrangements usually result from three of fewer breaks. We report a complex intrachromosomal rearrangement resulting from five breaks in one chromosome 10 of a phenotypically normal father of two developmentally delayed children. GTG-banding analysis of the father's rearranged chromosome 10 suggested in initial pericentric inversion followed by an insertion from the short arm into the terminal band of the long arm [der(10) (pter-->p13::q21.2-->p12.2::q22.1::-->q26.3::q22.1-->q 21.2::p12.2-->p13::q26.3-->qter)]. To our knowledge, this rearrangement is the most complex ever reported in a single chromosome. Both children inherited a recombinant chromosome 10 with loss of the insertion and the segment distal to it [rec(10)der(pter-->p13: :q21.2-->p12.2::q22.1-->q26.3:)]. Mechanisms for both rearrangements are proposed.

We report on a 5-month-old boy with a de novo interstitial deletion of the proximal short arm of chromosome 8 (p21p11.2). He manifested bilateral cleft lip and palate, and apparent hypogonadism. Four previous case reports with similar deletions (p11.1p21) were associated with hypogonadotropic hypogonadism [Beighle et al., Hum Genet 38:113-121, 1977] and hereditary spherocytosis (HS) [Chilcote et al., Blood 6:156-159, 1987; Kitatani et al., Hum Genet 78:94-95, 1988; Lux et al., Nature 345:736-739, 1990]. Our patient has no demonstrable red blood cell abnormality, suggesting that the gene for HS is located in the region 8p11.1 to 8p11.2.

e22171 Background: We previously have studied differentially microRNA expression levels related to hormone refractory and sensitive prostate cancer cell lines (VCap and LNCap respectively). Now, we investigated circulating miRNAs differentially expressed between metastatic prostate adenocarcinomas (mPA) and healthy controls (HC) that may serve as novel diagnostic and/or prognosis markers. Methods: Using SYBR-green-based custom microRNA RT-qPCR arrays technology (Exiqon), we compared the expression levels of miRNAs in blood samples from 18 HC and 48 mPA. Results: Among a panel of 92 candidates we found 41 underexpressed (UE) and 8 overexpressed (OE) miRNAs. The level of significance was p

e15542 Background: Renal cell carcinoma (RCC) is known to be resistant to chemotherapy. The need for the identification of biomarkers capable of determining RCC prognosis factors and metastatic potential that are obtainable from non-invasive or minimally invasive techniques are desirable. The purpose from this analysis was to derive predictive models which could predict more accurately than any one factor alone. Methods: By tissue micro-array (TMA) technology we assessed 29 markers involved in the cascade of events leading to the formation and progression of this disease. The ability of multivariate logistic regression to improve the diagnostic accuracy for RCC by combining determinations of proteins markers and pathological variables was evaluated. Univariate analysis was used to select factors (p

448 Background: Many studies have demonstrated genetic and environmental factors lead to renal cell carcinoma (RCC) occurring during a protracted period of tumorigenesis. It seemed suitable identify and characterize potential molecular markers which might provide rapid and effective possibilities for early detection of RCC. The purpose from this analysis was to derive predictive models which could predict more accurately than any one factor alone. Methods: We assessed using quantitative real-time PCR (qPCR) with SYBR Green the profile of 32 predictive markers involved in the cascade of events leading to the formation and progression of this disease to evaluate their involvement in oncogenesis. RNA of quality was obtained in 90% of samples (N=80, 52/80 clear RCC, 6/80 papillary RCC and 14/80 chromophobe RCC) to carry out the study of gene expression. GenEx software was used for qPCR data processing and analysis. The potential correlation between mRNA expression and the pathological features of the study subjects was assessed by Pearson Chi-squared. Linear rather than logistic regression models were used (SPSS statistics 21.0 software). Additionally, a knowledgebases of biological pathways, Reactome and Snow (Babelomics), were used to superimpose our quantitative expression data. Results: The best gene predictors related to different pathological variables (histological type, tumor size, Fuhrman grade, number of involved nodes, renal pelvis invasion, lymphatic vessels invasion, and rupture of the renal capsule): Hif1-α (p < 0.001), Hif1-β (p = 0.035), DLL1 (p = 0 .012), DLL3 (p< 0.001), c-Kit (p = 0.002), PDGFR-β (p =0.002), PDGFR-α (p = 0.026), Bax-β (p = 0.005), Survivin (p = 0.028), Notch1 (p = 0.006), Notch2 (p = 0.007), Notch3 (p = 0.016), Notch4 (p = 0.005), EGFR (p = 0.029), Glut3 (p = 0.026), Glut5 (p = 0.036), FH (p = 0.018), CA9 (p = 0.030), and VHL-1(p = 0.015). Conclusions: Our data suggested that altered expression of certain members involved in the main pathways which feed RCC and their downstream targets analyzed together could improve the diagnostic accuracy for renal cancer by combining determinations of mRNA markers.

e15077 Background: Renal cell carcinomas (RCC) constitute a morphologically and genetically heterogeneous tumor.Wnt proteins are a large family of secreted glycoproteins that activate intracellular signal transduction pathways to control a wide variety of cellular processes such as determination of cell fate, proliferation, migration, and polarity.Wnt signalling may be playing a central role during RCC development. Methods: To characterize Wnt signalling activity in RCC, we gathered 48 tumor samples from patients diagnosed with metastatic RCC between years 2006 and 2011. Expression levels of Wnt family members (Wnt1, Wnt2, Wnt3a, Wnt4, Wnt5a, Wnt7a, and Wnt9a) were examined by real-time PCR and immunohistochemistry in tumor and normal tissue from removed kidneys. Results: Wnt1 and Wnt5a were up-regulated in clear cell RCC being their relative expression 2.4.103 (Wnt1) and 4.102 (Wnt5a) times higher in renal tumor tissue than in normal tissue. One-way ANOVA test showed Wnt3a and Wnt4 as the best protein markers to distinguish clear cell RCC (F=3.95, p=0.010 and F=3.05, p=0.029, respectively). Wnt1 and Wnt7a were over-expressed in those cases which had undergone nefrectomy (Mann-Whitney U test p=0.065 and p=0.056 respectively). Finally, Wnt2 protein level was associated with those patients with high risk prognosis (Kruskal-Wallis test p=0.001). Conclusions: Our results indicate that Wnt3a and Wnt4 are potentially useful immunohistochemical markers for the differential diagnosis between clear cell RCC and other subtypes of RCC and also suggest a role for Wnt2 as a high risk prognosis marker.

e15076 Background: The characterization of factors involved in the etiology of renal cell carcinoma (RCC) may lead to the identification of new targets for therapy and prognostic markers. Recent data indicate that Notch family members can be expressed by malignant RCC cells and this expression may have potential use as a strong prognostic factor. The aim of this study was to investigate the potential of Notch family members as prognostic markers for stage IV patients. Methods: We analyzed, by means of immunohistochemistry and quantitative polymerase chain reaction, protein and mRNA levels of 4 Notch family members in tumor specimens obtained from 48 patients with stage IV RCC. Clinicopathological data for these patients were evaluated. A 34% of the population had hypertension, 7.3% had hypothyroidism, 19.5% were diabetic, 7.7% prediabetic, and 22% showed hyperlipidemia. RCC specimens included clear cell RCC: 34; papillary: 3; mixed: 1; undifferentiated: 1; hypernephroma: 1.The association among biomarkers and clinicopathological factors were explored using One-Way ANOVA test and non-parametric statistics, Mann-Whitney U and Kruskal-Wallis tests. Results: Relative expression of Nocth3 and Notch4 transcript levels were elevated 25-fold in clear cell RCC tumors when compared with normal kidney tissues. Notch1 and Notch2 were overexpressed at protein level in clear cell RCC (p>0.05). Those renal tumors with right localization showed highest levels of Notch4 protein (p=0.096). Notch2 overexpression was associated with patients who had undergone nefrectomy (p=0.038). Finally, Bonferroni post-hoc statistic test showed us a statistically significant association (p = 0.020) between Nocth1 protein expression levels and patients with medium risk prognosis. Conclusions: These results show a deregulated expression of Notch receptors in RCC and suggest a role for this pathway in the progression of this tumoral pathology.

e15079 Background: Many studies have demonstrated genetic and environmental factors lead to renal cell carcinoma (RCC) occurring during a protracted period of tumorigenesis. It seemed suitable identify and characterize potential molecular markers appearing during of tumorigenesis which might provide rapid and effective possibilities for early detection of RCC. Methods: We assessed using quantitative real-time PCR (qPCR) the profile of 32 markers involved in the cascade of events leading to the formation and progression of RCC: invasiveness, angiogenesis and antiapoptotic mechanisms. RNA of quality was obtained in 90% of samples (N=80, 64/80 clear RCC, 8/80 papillary RCC and 8/80 chromophobe RCC) to carry out the study of gene expression. Nonparametric tests (Pearson's correlation coefficient test and Spearman's rho test, Kruskal — Wallis and Mann — Whitney) and Bonferroni post-hoc test were applied for statistical data analysis. Additionally, a knowledgebase of biological pathways, Reactome, was used to superimpose our quantitative expression data. Results: Statistical analysis comparing medians and means showed the following candidates as the best gene predictors related to different pathological variables (histological type, number of involved nodes, renal pelvis invasion, lymphatic vessels invasion and rupture of the renal capsule): Hif1-α (p

405 Background: MicroRNAs (miRNAs), a class of small RNAs, have been intensely investigated recently because of their important regulatory role in gene expression. Circulating miARNs are potential biomarkers for cancer not only abundant in blood, but also very stable, which are important prerequisites as clinical biomarkers. Detection of circulating tumor cells (CTC) may provide diagnostic and prognostic information in genitourinary renal tumors. The aim of this work is identify miRNAs potentially useful for CTC detection in blood samples from patients with renal tumors to assess their potential clinical significance. Methods: We examined blood levels of three miRNAs, let-7c, miR-200c and miR-31 in 48 patients with renal tumors with locally advanced or metastatic disease and 18 healthy persons using quantitative real-time PCR. The clinicopathologic characteristics of the population under consideration include among other: sex, age, tumor location, histological type, Fuhrman grade, tumor size, number of involved nodes, tumor stage, time to progression and overall survival. Results: We found that the median levels of miRNAs, let-7c, miR-200c and miR-31 were significantly higher in patients with renal tumors than those in healthy controls (p=.002, p=.000 and p=.004, respectively). Receiver-operator characteristic (ROC) curve analyses indicate that these blood miRNAs may be useful markers for discriminating patients with renal tumors from healthy controls (AUC=.750, AUC=.853 and AUC=.738, respectively). Conclusions: miRNAs in the circulation are relatively stable, very accessible, low invasive and easily testable biomarkers. Our results suggest let-7c, miR-200c and miR-31 as novel stable blood-based markers for renal cancer detection. This work was supported by Grants PI07/0477 from Fondo de Investigaciones Sanitarias (FIS), Instituto de Salud Carlos III.S. Díaz Prado is supported by an Isidro Parga Pondal research contract by Xunta de Galicia (A Coruña, Galicia, Spain). Cancer research in our laboratory is supported by the “Fundación do CHU A Coruña”.

We present a patient with severe idiopathic aplastic anemia with no previous chromosomal abnormalities who developed trisomy 21 and monosomy 7 during treatment with intravenous (i.v.) cyclosporine. The abnormal karyotype disappeared when the drug was changed to the oral form. This cytogenetic aberration, previously unreported in association with cyclosporine, may reflect either a direct drug effect or the emergence of a hidden myelodysplastic cell clone subject to preferential survival during immunosuppression.

Cytogenetic studies of the peripheral blood and marrow of a patient with chronic granulocytic leukemia in blastic transformation revealed the following abnormalities: 1) a Philadelphia chromosome with the usual 9:22 translocation; 2) a three break rearrangement (insertion) of chromosome #11; and 3) a deletion of the short arm in chromosome #16. Clinical and hematologic remission developed after three courses of intermittent chemotherapy with transient but complete bone marrow aplasia. Studies performed during remission revealed two cellular clones. One had the same karyotype already described and the second showed only a Ph1 chromosome. We postulate that the pretreatment clone evolved from Ph1-positive cells in this patient with chronic granulocytic leukemia during a short, undiagnosed stable phase. Treatment allowed the reappearance of the Ph1-positive clone. The second abnormal clone was apparently partially eradicated but normal cells did not repopulate the bone marrow, suggesting that either the Ph1-positive cells were greatly resistant to therapy or that there were no remaining normal myelogenous stem cells. In addition, we found that there have been few reports of abnormalities involving chromosomes #11 and #16, and that an inserting chromosome #11 has never been reported.

Herein we give clinical support to the theory of a common origin of multiple endocrine adenomatosis. Accurate clinical evaluation of a patient suspected to have endocrine neoplasia requires the search for a much wider range of potential endocrine tumors. Such an approach is now made possible by the radioimmunoassay. Chromosome patterns and histocompatibility antigens are reviewed in a family with the disease.

Activity and association patterns of nucleolar organizer regions (NORs) were studied in cultured blood lymphocytes from six members of a family, three of whom carried a doubly satellited marker. The marker consisted of a centromere with two sets of satellites and stalks which showed consistent NOR activity. The children with the marker had significantly higher frequencies of NOR activity and satellite association than their non-carrier siblings. The acrocentrics of the children with the marker did not appear to have decreased association tendency.

Associations between nucleolus organizer regions (NORs) and non-acrocentric chromosomes were scored in 2,800 metaphase spreads from PHA-stimulated lymphocyte cultures (48 h) from 14 individuals. The preparations were both silver stained and C-banded. In order to calculate the expected values for associations, the ratio of heterochromatin length to euchromatin length was established for each subject. Individual C-band lengths and centromeric lengths were also determined. When silver connective (SC) associations with heterochromatin were compared to SC associations with euchromatin, the number of associations with heterochromatin was significantly greater than expected (P < 0.000001) for each subject. The SC associations were not distributed randomly over the heterochromatin of the non-acrocentrics. Chromosomes 1 and 2 had significantly more than expected. Chromosomes 17, 18, 19, 20, and the Y had fewer than expected. NOR associations with euchromatic segments also showed a nonrandom pattern of distribution.

A 46,XX,inv(12)pat/47,XX,i(Xq),inv(12)pat was diagnosed prenatally in a 36-year-old woman whose husband was a known carrier of a pericentric inversion of chromosome 12. The diagnosis was confirmed in fetal tissue. Terminal bromodeoxyuridine (BrdU) labelling demonstrated that in the line with 46 chromosomes one X was late replicating, while one X and the i(Xq) were late replicating in 100% of the cells with 47 chromosomes. We present the first case of this type of sex chromosome mosaicism. Genetic counseling presented difficulties since it was not possible to predict the fetal phenotype.