ALBERT

Description: Background: Gene expression profiles have been associated with prognosis in myelodysplastic syndromes. WT1 is a tumor-suppressor gene coding for a transcription factor located on chromosome 11p13, which was originally identified for its involvement in the pathogenesis of the Wilms’ tumor. In normal bone marrow, WT1 expression is low or undetectable, whereas it is aberrantly expressed in hematological malignancies. Evidence indicates that WT1 is important in the lineage-specific differentiation of hematopoietic cells and leukemogenesis. In myelodysplastic syndromes (MDS), WT1 expression has prognostic significance: it is directly correlated with the type of MDS, with IPSS score and with disease progression. Recent data demonstrate that WT1 is a potent activator of the EPO gene under normoxia and it is suggested that WT1 may regulate paracrine EPO synthesis in a tissue-specific manner. Bmi-1 is a transcriptional repressor gene which may be expressed restrictedly in stem cells and progenitors and is required to regulate the adult self-renewing hematopoietic and leukemic stem cells. It appears that it also plays an important role in providing cells the potential for proliferation. A number of reports on Bmi-1 provide perspectives on the close association of its expression with the progression of hematopoietic malignancies. Furthermore, flow cytometry has shown that Bmi-1 positivity in CD34+ cells is positively correlated with IPSS score. Introduction: We have designed a study to evaluate changes in gene expression profiles of bone marrow mononuclear cells of primary low and intermediate-1 IPSS risk MDS patients receiving erythropoetic growth factors (darbepoetin or high-dose rHuEpo alpha). Associations with response, changes in Hb, in percentage of CD34+ and apoptotic cells are evaluated. We present preliminary results in 6 patients. Methods: Bone marrow samples were obtained before and after 12 weeks of treatment. Mononucleated cells were cryopreserved and later thawed for total RNA extraction and cDNA synthesis. Gene expression profiling and the expressions of WT1 and Bmi-1 by RT-PCR were evaluated. Results: Baseline median Hb was 9,5 g/dL (interquartile range 8,7 10,1). Baseline mean Bmi-1 was 14011 (± SD 3927). All patients had a major erythroid response to treatment. Preliminary results demonstrate a significant decrease in WT1 from baseline median 1635 (interquartile range 1251–2150) to 1192 (interquartile range 1005–1365, P=0.046). Fig. 1. Changes in WT1 expression during therapy. Fig. 1. Changes in WT1 expression during therapy. Discussion: Though few patients have yet been studied, it is suggested that WT1 decreases in patients responding to erythropoetic growth factors. Further evaluation with stratification for erythropoetic response in the extended study may furnish novel associations between gene expression, erythropoetic growth factors and prognosis in MDS patients.

Description: The prognostic significance of the response to initial prednisone treatment in adult ALL has been recently emphasized. Prednisone response is usually defined on the basis of the peripheral leukemic blast count. The threshold value for the defintion as good or poor prednisone response is 1000 blasts/mmc on day 8 of prednisone pre-treatment. The drawback of this definition is the difficulty of classifying patients with less than 1000 blasts at diagnosis. In the LAL2000 GIMEMA study we therefore evaluated whether the blast reduction rate, which is not affected by the initial blast level, could be a factor with comparable prognostic value. The protocol design provided a 7-day (−6 to 0) pre-treatment phase with an escalating dose of prednisone up to 60 mg/sqm. On day 1 before starting the induction the response was assessed both according to the absolute blast count (〈 versus ≥ 1000/mmc) (criterion 1) and according to the blast reduction rate ≥ 75% (criterion 2) in the peripheral blood. The induction included high dose Daunorubicin; for patients in complete remission (CR) this was followed by consolidation with high dose ARA-C, chemo and radio prophylaxis of the central nervous system, and periodical reinduction over a three years maintenance period. Patients with adverse cytogenetic features [i.e. t(9;22), t(4;11), t(1;19)] who achieved a CR were treated according to the HAM protocol that included high dose ARA-C and Mitoxantrone followed by Imatinib for Ph+ ALL and by allogeneic or autologous hemopoietic stem cells transplantation for the others. Between September 2000 and December 2003 a total of 368 patients were evaluable for response to induction. The median age was 35 years (15–60) and median WBC count 15′109/L (0.3–872); 72 (20%) were T ALL and 121 (33%) had cytogenetic high risk features (104 (86%) Ph+, 4 (3%) t(4;11) and 13 (11%) t(1;19)). Eighty-seven percent of the patients were evaluable for response to steroid pre-treatment: ’responders’ were 75% according to criterion 1 (blast

Description: Epigenetic silencing of tumor suppressor (TS) genes is a hallmark in human leukemias, particularly through DNA methylation. Cyclin-dependent kinase inhibitors (CKI) are, among other genes, frequently found methylated in their promoter region. This epigenetic modification has been described also in acute lymphoblastic leukemia (ALL). However, the relationship between aberrant DNA methylation and protein expression of TS genes has not yet been extensively evaluated in adult ALL series. The aim of this study was to analyze in primary cells from newly diagnosed adult ALL patients, uniformly treated according to the LAL2000 GIMEMA protocol, the promoter methylation status of p73, p21, p15 and p16, evaluating in addition the p21, p15 and p16 protein expression. The DNA methylation status of promoter regions was investigated, according to cell availability, using a widely accepted method based on bisulfite modification of DNA, followed by methylation-specific PCR assay (MSP). Protein expression was evaluated by Western blot. Normal peripheral blood lymphocytes, as already described, resulted unmethylated for p73, p21, p15 and p16, and did not express the p21, p15 and p16 proteins. In ALL patients, in contrast, only the p21 promoter region was found constantly unmethylated. The p15, p16 and p73 promoter genes were found methylated in 15/37 (40.5%), 8/43 (18.6%) and 9/36 (25%) patients, respectively. Only 2/23 cases (8.6%) resulted simultaneously methylated for p15, p16 and p73. The p21 and p15 protein expression was found in 28/85 (32.9%) and 44/85 cases (51.8%), respectively. The p16 protein, in contrast, was never expressed. The p16 methylation was associated with the T-ALL (P=0.005) phenotype and with higher white blood cell (WBC) counts (P=0.027). Resistance to spontaneous induction of apoptosis was significantly associated with p21 protein expression (P=0.019) and its co-expression with p15 (P=0.049). Achievement of CR was not influenced by gene methylation status, nor by single protein expression. Interestingly, the co-expression of p15 and p21 was associated with failure to induction treatment: only 6/63 (9.5%) patients co-expressing p15 and p21 obtained a CR (P=0.027). Multivariate analysis confirmed the unfavorable role of this protein co-expression (P=0.059) on CR achievement. In contrast, once patients achieved remission, p21 protein expression was associated with a prolonged DFS, as confirmed by multivariate analysis for DFS (P=0.039). In conclusion, p15 and p21 protein expression plays an unfavorable prognostic role in adult ALL patients independently of the p73, p21, p15 and p16 gene promoter methylation status.

Description: Dasatinib was approved for use in the treatment of patients (pts) with chronic myeloid leukaemia (CML) and resistance to Imatinib. We applied a rescue treatment based on Dasatinib therapy to achieve a pharmacological immunomodulation in a setting of CML-relapsed allogenic stem cell transplantated (A-HSCT) pts. Patients were required to have both resistance to Imatinib and unresponsiveness to cellular therapy such as Donor Lymphocyte Induction (DLI). We hypothesized that Dasatinib could potentially improove the disease by immunomodulatory action. Primary aim of this therapeutic design was to address, in single institution trial, therapeutic force of a innovative pharmacologic strategy to induce the cytogenetic response followed by DLI. Therefore, we investigated Dasatinib ability to achieve immuno-effects by targeting key mediators of Th1, Th2, and Treg response. Biological effects were examined on conventional diagnostic parameters such as haematological chimerism, cariotype and Bcr-Abl gene transcript. Herein, we present interim results of a pilot group of 3pts. Patients received dasatinib 70 mg twice daily(140 mg total daily dose). Dose modifications were allowed for the management of toxicity. Treatment was performed until complete cytogenetic, molecular response and haematological full donor chimerism. Materials and Methods: To investigate the immunological changes, we used a TaqMan® Low Density Array, based on comparative CTdd CT method on Applied Biosystems 7900HT, to perform relative quantification of cDNA derived from peripheral venous blood specimens harvested after DLI, before and after starting dasatinib therapy. Assumed that normal control values of all transcripts were = 1, we evaluated over or down regulation of gene expression profile (GEP) of a panel of 48 genes involved in immune response. Results: clinical changes after third month of dasatinib therapy. Case 1: responsive patient, maintained a mixed haematologic chimerism, but showed a complete cytogenetic and molecular remission. Following, patient restarted with DLI therapy. Case 2: responsive patient, showed nearly full-donor haematologic chimerism with complete cytogenetic and molecular remission. Case 3: patient no evalutable because brief treatment (only 1 month). Dasatinib caused early haematological toxicity. Patient maintained a low level of donor T cells with presence of Philadelphia chromosome associated to elevated p210 molecular signal. Gene expression profiles post-dasatinib therapy: According to in vitro experiments (Blood October 25, 2007), in all cases we observed a down regulation of IL-2 and IL-12B (Th1), IL-6 and IL- 18, IL-10 (Th2) cytokines and mediators of apoptosis such as EGR2, EGR1. By contrast, multiple pro-inflammatory factors were up-expressed: IFN-g, IL-17, IL-7. Only in case number 1, TNF-a and IFN-g molecular pathways were not influenced by the drug. In fact their elevated expression was preserved as compared to pre-dasatinib levels. Noterworthy among cases number 2 and 3 (with mixed chimerism), Dasatinib improved a marked inhibition of Th1 effectors in addition to down-regulation of several important molecular transcripts: SERPIN B3–B4, BCL2A1, SELP, PIAS 1, IRF8, IRF1, CCL7, CCL5, CXCL9, CCR4, ICAM. Regards to T regulatory cells, Foxp3 was strongly up-regulated in case number 1 and down-regulated in case2and 3. Conclusion: We think that Dasatinib represent a possibility of cure for for CML pts relapsed after A-HSCT and unresponsive to alternative treatments. For imatinib-resistant CML patients, such as in this study, there are few currently available effective options. The present results strongly emphasize the importance of immune response control to achieve the desired clinical effects.

Description: INTRODUCTION: Quality-of-life (QOL) is a patient-related outcome of increasing interest in onco-haematological setting. However, there are very few reports on QOL in elderly patients with Acute Myeloid Leukemia (AML). Prognosis of AML in elderly patients is still poor with a median survival of 9–12 months and less than 20% survival at 5 years. This is due to age-related factors and also to a higher incidence of poor-risk cytogenetics, multi-drug resistance and treatment-related mortality. The measurement of QOL at diagnosis may provide useful information regarding patient preferences and prognosis while followup measurements may indicate acceptance, adaptation and adverse effects of disease and therapy. METHODS: From 2/2003 and 2/2007, we included 113 elderly AML patients in a multicenter Italian 12-month observational study to evaluate the association of baseline QOL scores and their changes with disease factors, therapy and survival. Patients aged 〉 60 years (M/F 58/55, mean age 71.7 ± 5.9 yrs) from 4 Institutions with “de novo” AML according to WHO classification were evaluated from diagnosis prospectively up to 12 months. Two different questionnaires were employed: the EORTC QLQ-C30 and the QOL-E v.2. Therapeutic choice was not restricted by protocol and was given freely by the individual center. RESULTS: Forty-eight patients (43%) received intensive chemotherapy and 65 (57%) lowdose therapy or supportive care. Both age 〉 70 years (p=0.007) and concomitant diseases (p=0.031) had a significative impact on medical decision for palliative approaches. At diagnosis, general QOL was affected (median QOL-E standardized score 54, IQ range 47– 68, median EORTC global score 50, IQ range 42–67), loss of appetite was perceived by 75% of patients and QOL-E fatigue scores were low, indicating poorer QOL (median 45, IQ range 32–53). There was a significant correlation between fatigue and age, Hb levels and the duration of fever. In a multivariate regression model, both Hb and age indipendently predicted fatigue (linear R2 0.114, p=0.001, and linear R2 0.066, p=0,010). Most patients were given a good ECOG Performance Status (〈 2) that, interestingly, did not correlate with the perception of QOL, in particular physical and functional scores. QOL scores worsened after 1 month from diagnosis but patients surviving thereafter perceived an improvement in the following months. When correcting for the effects of age, concomitant diseases and changes in Hb values, fatigue improved in patients undergoing aggressive chemotherapy while it worsened in those on standard or supportive-care (p=0.004). Median overall survival was 49 weeks (95% CI 35–63). Patients receiving aggressive therapy had a longer survival with the median not reached at 60 weeks and 72% of patients surviving versus patients receiving palliative care (median 39 weeks, 95% CI 16–63, p 70 yrs (HR 2.4, 95%CI 1.2 – 5.0, p=0.02) and concomitant diseases (HR 2.0, 95% CI 1.1 – 3.8, p=0.044) had an independent negative prognostic impact on survival, as previously reported in many other studies. However, in order to evaluate the predictive value of QOL at diagnosis for survival, in a multivariate model corrected for age, concomitant diseases and treatment option, QOL measures that independently predicted survival were fatigue (p=0.003), global QOL (p = 0.004), physical QOL (p = 0.006) and functional QOL (p = 0.002). CONCLUSION, QOL seems to have an important role also in the elderly AML setting: though QOL is a highly subjective measurable value, we outline the role of QOL measures at diagnosis as a prognostic factor for overall survival and, thus, as a potential factor for treatment decision.

Description: Abstract 4848 Objectives: the prognosis of patients with cytogenetically normal acute myeloid leukemia (CN-AML) is highly variable and can be influenced by several clinical and biological variables. Nevertheless, some biological data may be conflicting and difficult to combine with the clinical ones. Methods: in order to propose a simple scoring system, we retrospectively analysed the clinical data of 337 patients newly diagnosed with CN-AMLs, aged less than 65 years, consecutively treated in eleven hematological Italian Centres from 1990 to 2005. Two hundred nineteen patients (65%) received a fludarabine-based induction regimen. All the other patients received a conventional induction regimen, including cytarabine, one anthracycline with or without etoposide. Univariate and multivariate analysis on event free survival and overall survival (EFS and OS) were performed. Patients addressed to allogeneic stem cell transplantation were censored at the time of transplant. Factors found to be significant in univariate analysis were tested in multivariate analysis. A numerical score was derived from the regression coefficients of each independent prognostic variable. The Prognostic Index Score (PIS) for each patient was then calculated by totalling up the score of each independent variable. Patients could thus be stratified into low-risk (score = 0–1), intermediate-risk (score = 2) and high-risk group (score grater than 3). The score obtained in this group of patients (training set) was then tested on 193 patients with newly diagnosed with CN-AMLs, aged less than 65 years, enrolled in the GIMEMA LAM99p clinical trial (validation set). Results: the clinical variables that were independent prognostic factors on EFS in the training set of patients were: age 〉 50 yrs (regression coefficient: 0.39, HR 1.5, score = 1), secondary AML (regression coefficient: 0.90, HR 2.5, score = 2) and WBC 〉 20 × 10^9/L (regression coefficient: 0.83, HR 2.3, score = 2). For what concerns the OS, the same variables showed the followings statistical data: age 〉 50 yrs (regression coefficient: 0.48, HR 1.6, score = 1), secondary AML (regression coefficient: 0.99, HR 2.7, score = 2) and WBC 〉 20 × 10^ 9/L (regression coefficient: 0.87, HR 2.4, score = 2). In the training set of patients, the median EFS was 22, 12 and 8 months in the low, intermediate and high-risk group (p

Description: Abstract 4974 Introduction: Primary immune thrombocytopenia (ITP) is an acquired immune-mediated disorder characterized by isolated thrombocytopenia, defined as a peripheral blood PLT count 60 years of age, those with systemic symptoms or abnormal signs or in some cases where splenectomy is considered (Provan et al, Blood, 2009). Methods: We performed a retrospective chart analysis between January 2008 and June 2010 of all patients referred to our clinic for isolated thrombocytopenia but with a PLT count 100– 149 Gi/L. According to recent guidelines, these patients are not to be considered thrombocytopenic and do not require further investigation. The aim of the study was to evaluate the validity of omitting BM analysis in these cases. Results: Twenty-three cases (13 males/10 females) of mean age 58 ± SD 19 years were evaluated at our clinic for a PLT count below normal lab range values. At the time of evaluation none had bleeding symptoms. PLT counts ranged from 101 to 149 Gi/L, mean 123 Gi/L. Four patients had an enlarged spleen. After initial screening, 2 patients had a complex autoimmune disorder and 1 case had HCV hepatitis. The remaining 20 patients had a bone marrow (BM) aspirate performed: a diagnosis of myelodysplastic syndrome (MDS, WHO classification refractory thrombocytopenia) was obtained in 13 cases (65%) and BM biopsy was performed in 12, completed by cytogenetics in 9 cases (7 normal, 1 del20q, 1 –Y). Patients diagnosed with MDS were significantly older (66 ± SD 13 vs 47 ± SD 21 years, p = 0.017), but 4 cases (31%) were 〈 60 years of age (44, 49, 51 and 55 years of age, respectively). Conclusions: The most recent guidelines, which lower the PLT threshold to 100 Gi/L from 150 Gi/L for the investigation of causes of thrombocytopenia, reduce the diagnostic rate of MDS. In our retrospective review, 65% of patients would not have had an early diagnosis of MDS. Furthermore, BM aspirate should be considered irrespective of age, since one third of the patients in our case review had MDS with PLT 〉 100 Gi/L as a single cytopenia and age under 60. Disclosures: No relevant conflicts of interest to declare.

Description: Abstract 2927 Background: Immunological changes have a primary role in the initiation and progression of myelodysplastic syndromes (MDS). Cytokine levels, such as IL-7 and IFN-gamma, are associated with lower-risk disease. Treatment with lenalidomide has proven efficacy in red blood cell (RBC) transfusion-dependent lower-risk MDS patients with del(5q). Lenalidomide exerts anti-angiogenic, anti-proliferative, and pro-erythropoietic effects; in particular, it has been shown that lenalidomide inhibits the proliferation and function of T regulatory cells (Tregs). Finally, MDS patients undergoing lenalidomide treatment experience erythroid responses and suppression of the del(5q) clone. Aims: In a multicenter Italian phase II trial to evaluate safety and efficacy of lenalidomide in primary MDS patients with del(5q) and Low- or Int-1 risk IPSS, we investigated changes in the transcription of cytokines and their receptors during treatment. Methods: The starting dose of lenalidomide was 10 mg p.o once daily on a continuous daily schedule for a maximum of 12 months. Bone marrow (BM) aspirates were obtained on study entry and every 12 weeks. Assays were performed using TaqMan® Low Density Array Fluidic card (TaqMan® Human Array, Applied Biosystems, Foster City, CA, USA) based on Applied Biosystems PRISM® 7900HT comparative ddCT method, according to the manufacturer's instructions. Target gene expression levels were measured in triplicate and normalized against the expression of the 18S housekeeping gene from a BM pool of normal, healthy subjects at all timepoints. Median relative gene expression values in MDS patients were compared to healthy subjects, set as a value of 1. Results: We report data obtained at baseline and after 12 weeks. Informed consent was obtained in all patients. Twenty-seven patients (5 M, 22 F) were evaluated at baseline and after 12 weeks. Mean age was 72 ± 9 years. Mean Hb level was 8.5 ± 0.9 g/dL and 16 patients were RBC transfusion -dependent (requiring at least 4 RBC transfusions in the preceding 2 months). Seven patients had additional cytogenetic abnormalities. Twenty-one patients (80%) experienced erythroid responses by week 12. Significant variations in gene expression of cytokines and receptors were observed during treatment. Genes significantly regulated during lenalidomide treatment (P 〈 0.05) are shown in the Table. In particular, FAS, IL-7 and FOXP3 gene were generally under-expressed at baseline and significantly increased after 12 weeks. Accordingly, IL7R was over-expressed in all patients at baseline and its expression was significantly reduced during treatment. Furthermore, IFN-gamma expression increased during therapy. Summary: The protein encoded by FAS gene is a member of the TNF-receptor superfamily and its interaction with its ligand leads to apoptosis. Interleukin (IL)-7 is an essential cytokine that promotes the proliferation and survival of B- and T-lymphocyte progenitors. The IL7R gene on chromosome 5 (5p13) codifies for the IL7 receptor, which blocks apoptosis during differentiation and activation of T lymphocytes. It functions, in part, through the induction of the expression of the antiapoptotic protein Bcl-2. The protein encoded by the FOXP3 gene is a member of transcriptional regulators. Defects in this gene are the cause of X-linked autoimmunity-immunodeficiency syndrome. The results of the present study indicate that lenalidomide may act through immunological changes. Further detailed analyses in these patients may provide new insights into the pathogenesis of MDS with del(5q) and the long-term effects of lenalidomide treatment on immunological changes in BM cells. Disclosures: Oliva: Celgene: Consultancy.

Description: Abstract 3078 Chronic myeloproliferative neoplasms (CMNs) include Polycythemia Vera (PV), Essential Thrombocythemia (ET), and Primary Myelofibrosis (PMF). So far limited studies of familial clusters of CMNs have been reported.Familial chronic myeloproliferative neoplasms are defined when in the same pedigree at least two relatives have CMNs. Familial CMNs should be distinguished from inherited disorders with Mendelian transmission, high penetrance and polyclonal haematopoiesis named ‘hereditary erythrocytosis' and ‘hereditary thrombocytosis'. Recently a 5- to 7-fold higher risk of MPN among first-degree relatives of patients with MPNs was reported. These findings support the limited studies suggesting a familial clustering in MPNs. The analysis of mutations of JAK2 and MPL may improve our ability to identify these conditions. In a consecutive series of patients observed in our Institution from January 2000 to June 2010, we found that among 460 patients with sporadic CMNs and 94 Ph1 positive chronic myeloid leukemia (CML), the prevalence of familial cases was 4%.With 22 pedigrees, 44 patients (8%) were identified with two relatives affected. Familial CMNs were 11 PV,14 ET,7 PMF, 5 CML respectively, while sporadic cases were 96 PV,204 ET,115 PMF and with other 45 CMNS not furtherly classified. As far as the distribution of the different CMNs within the familial cluster, We observed that only in 4 of 22 families (18%) all the affected relatives were diagnosed with the same disease (homogeneous pattern: PV one family and ET three families), whereas 14 families exhibited a mixed distribution among PV, ET and PMF. 8 families exhibited CMNs associated with other hematological disease such as chrocic lymphocytic leukemia (CLL), acute myeloid leukemia (AML), chronic myeloid leukemia (CML), myelodisplastic syndrome (MDS). Among this, 6 families presented a first or second degree of relationship of first and second generation. In 10 cases the relatives were brothers, affected by familial CMNs with a prevalence of PV and TE clinical phenotype at diagnosis.According to JAK2 (V617F) mutational status, analyzed in 30 out of 44 patients, 19 patients showed a positivity pattern, while 18 families showed a heterogeneous pattern; they included both JAK2 (V617F) -positive and JAK2 (V617F)-negative patients. Among the 19 patients with JAK2 (V617F) positivity, the distribution of positivity according to the diagnosis was 100% of PV, 45% of ET and 55%of PMF; homozygosity was present only in PV cases. In our series, only two members of the same family were affected by familial CMNs. Finally it should be noted that in our series of familial cases clinical presentation, therapeutic approach and type and severity of complications were comparable to that of sporadic cases. In conclusion, the present study indicates the relevant possibility of familial CMNs, thus suggesting the opportunity of a detailed family history as part of the initial work-up of patients with CMDs; in addition it also suggests the usefulness of an accurate biological study. Disclosures: No relevant conflicts of interest to declare.

Description: Abstract 4040 Multiple myeloma (MM) is a clonal proliferation of malignant plasma cells (PCs) characterized by a marked genomic instability. Primary plasma-cell leukemia (pPCL) is an aggressive, rare variant of plasma cell dyscrasia characterized by extra-medullary proliferation of PCs, high genomic instability and very poor prognosis. To date, global genomics studies in pPCL are still limited. Highly purified PCs were obtained from 17 previously untreated pPCL patients, recruited in an open-label, exploratory, single-arm, two-stage study from the GIMEMA myeloma network aimed at the evaluation of safety and antitumor activity of the immunomodulatory agent lenalidomide in combination with low dose dexamethasone in previously untreated pPCL. All the samples were characterized for the main chromosomal aberrations by Fluorescence in-situ hybridization (FISH). Specifically, 13q and 17p deletions have been identified in 12/17 (70.6%) and 8/17 (47.1%) cases, respectively; the presence of t(11;14) was found in 4/17 patients (23.5%), t(4;14) in only 1/17 cases (5.9%) and MAF translocations in 8/17 (47.1%). To further investigate the genomic complexity of pPCL, we analyzed a subset of 13 samples by means of an integrative approach using different high-throughput microarray technologies: Human Mapping 250K Nsp SNP-array (Affymetrix) for the detection of copy number alterations (CNA), Gene 1.0 ST array (Affymetrix) for transcriptional profiling and miRNA Microarray v2 (Agilent) for the global miRNA expression. SNP-array data were concordant with FISH results for the detection of the main chromosomal aberrations, i.e. 13q (10/13=77%) and 17p (7/13=58.3%) deletions. The most recurrent CNA specifically identified by SNP-array was represented by 1q gain (8/13=61.5%); in addition. losses involving chromosomes 1p (5/13=38.5%), 8p (4/13=30.8%), 14q (5/13=38.5%), 16q (5/13=38.5%), gains affected 7q (4/13=30.8%) and 19p (4/13=30.8%) and one amplification at 17q21 in 6/13 pPCL (46.2%) were detected. One case displayed a near tetraploid karyotype and, interestingly, another one showed a hyperdiploid pattern. Mapping information was integrated with the gene expression and miRNA profiles of the tumor samples. A non-parametric analysis (Kendall's tau correlation at a p-value