Poster Session, Saturday 28 January 2017 Abstracts S143 1396 POSTER Effectiveness of a relaxation intervention technique (progressive muscle relaxation and guided imagery techniques) to reduce anxiety of parents of hospitalized children T. Tsitsi 1 . 1 Cyprus University of Technology, Nursing, Limassol, Cyprus Introduction: The diagnosis of Malignancy in a child is a source of intense stress in patient and their family. Anxiety, depression, anger and low self- esteem are some of the emotional reactions experienced by parents in response to the fear of loss and future relapse. To investigate the effect of Progressive Muscle Relaxation (PMR) and Guided Imagery (GI) interventions, in reducing anxiety levels among parents of children diagnosed with any type of malignancy receiving active treatment at the Paediatric Oncology unit. Material and Method: A parallel group randomized non-blinded control trial, conducted between April 2012 to October 2013, at two public hospitals in Cyprus and Athens. Fifty four parents of children hospitalized with malignancy were randomly assigned to the intervention (PMR and GI) (n = 29) or control group (n = 25). Parents were randomized to receive in individual sessions, a script process of Progressive Muscle Relaxation and Guided Imagery techniques, once a week for 25 minutes in a private room at the Paediatric Oncology unit of the hospital, for three weeks. Parents randomized to the control group did not receive any additional intervention beyond the usual psychological support. The outcome variables of the study were changes in anxiety levels (Hamilton’s Anxiety Scale and physiologic measures of anxiety (blood pressure, pulse, skin temperature) and mood (Questionnaire of Profile of Mood States Brief), at 3 weeks’ time. Results and Discussion: Significant differences of mean were found in the Hamilton Anxiety Scale between T0 and T1 in the two groups. In the intervention group the correlation coefficient of the score on the Hamilton anxiety scale in the T0 and in the T1 measurement was r = 0.843 (p < 0001) and in the control group r = 0.847 (p < 0001). In the POMs Brief scale, there was a statistically significant difference in tension at the T0 measurement and T1 (p = 0.027) and the parents in T1 measurement in the intervention group were significantly less sad (p = 0.001), and significantly less tense and anxious (p = 0.031). The findings showed the beneficial nature of relaxation to reduce anxiety and improve mood in parents of children with malignancy. This research was supported by the acceptance, the effectiveness and the impact of these techniques to reduce the anxiety of the parents of children with malignancy. The study was registered in the ClinicalTrials.gov:NCT01590524. No conflict of interest. 1397 POSTER Integrative analysis of bone marrow disease in neuroblastoma patients by DNA, RNA and protein markers A. Druy 1 , E. Shorikov 2 , G. Tsaur 3 , A. Popov 4 , S. Tuponogov 3 , A. Zaychikov 3 , L. Saveliev 5 , L. Fechina 6 . 1 Federal Research and Clinical Center for Pediatric Hematology- Oncology and Immunology, Laboratory of Cytogenetics and Molecular Genetics, Moscow, Russian Federation; 2 Morozov Children’s City Hospital, Pediatric Oncology, Moscow, Russian Federation; 3 Regional Children’s Hospital, Pediatric Oncology and Hematology Center, Ekaterinburg, Russian Federation; 4 Federal Research and Clinical Center for Pediatric Hematology- Oncology and Immunology, Cell Immunology and Immunogenesis Laboratory, Moscow, Russian Federation; 5 Ural State Medical University, Chair of Laboratory Medicine, Ekaterinburg, Russian Federation; 6 Research Institute of Medical Cell Technologies, Laboratory of Cellular Therapy of Oncohematological Disorders, Ekaterinburg, Russian Federation Background: BM involvement detection in neuroblastoma is crucial for staging; residual tumor cells (RTC) could define incomplete tumor response and be associated with unfavorable outcome. Methods: Expression of 4 tumor-associated genes (TAGs): PHOX2B, TH, ELAVL4, GD2 was analyzed in 26 intact BM samples, in discovery (331 BM samples from 57 patients) and validation (311 samples from 55 patients) cohorts. Threshold levels of expression established by ROC-analysis were applied for overall correct prediction (OCP) computation. In 239 BM samples from 64 patients methylated RASSF1a gene was analyzed. BM of 51 patients was subjected for analysis by flow cytometry detecting cells with tumor-associated immunophenotype (CD81+CD56+CD9+GD2+CD45−). All patients were treated according to NB2004 protocol. Event-free (EFS) and overall survival (OS) were calculated with median of follow-up 36.1 months. Results: Neither PHOX2B, TH expression nor mRASSF1a were detected in the intact BM, expression of ELAVL4 was revealed in 20, GD2 − in 15/26 samples. In the discovery cohort OCP values for PHOX2B, TH, ELAVL4, GD2 achieved 0.994, 0.952, 0.828, 0.767. In validation − 0.997 for PHOX2B, 0.939 for TH. Presence of PHOX2B/TH expression in BM at primary diagnostics decreased survival (table). Predominance of PHOX2B expression over TH >1.68 in the BM had adverse prognostic significance: EFS 0 vs 56%(12) p = 0.017, OS 0 vs 72%(11) p = 0.006. High-risk patients free from RTC during the induction treatment had superior survival. Persistence of RTC until the completeness of the induction chemotherapy was fatal. Positivity of BM for PHOX2B/TH before stem cells (SC) apheresis had strongly negative prognostic impact irrespective of CD34+ selection. Marker Survival rates, % (standard deviation, %) EFS OS + − p + − p Primary diagnostics PHOX2B/TH 41 (10) 81 (6) <0.001 49 (11) 87 (5) <0.001 mRASSF1a 32 (15) 77 (7) 0.003 56 (16) 80 (7) 0.085 Flow cytometry 28 (9) 84 (7) <0.001 36 (11) 88 (7) <0.001 PHOX2B/TH During induction chemo 21 (17) 50 (20) 0.022 21 (17) 67 (19) 0.044 After induction chemo 0 51 (18) 0.013 0 64 (17) 0.012 Before SC apheresis 0 47 (12) 0.040 0 61 (13) 0.028 +, positive; −, negative. 69 of 239 BM samples were positive for mRASSF1a, concordance rate with TAG expression achieved 80.3%. Prognostic impact of mRASSF1a presence in the BM of primary patients was less than PHOX2B/TH. Among 51 patients analyzed by immunophenotyping, 25 harbored BM infiltration and had inferior survival. Results of flow together with MYCN amplification and stage 4 disease demonstrated independent prognostic significance in the multivariate analysis. Concordance of flow results with TAGs expression was 78.8%. Conclusion: Expression of PHOX2B/TH and flow are the most appropriate techniques for BM involvement detection with significant prognostic impact. No conflict of interest. 1398 POSTER SNPS in microRNAs associated with methotrexate plasma levels in Spanish children with acute lymphoblastic leukemia M. Umerez 1 , A. Gutierrez-Camino 1 , I. Martin-Guerrero 1 , N. Garcia De Andoin 2 , A. Sastre 3 , A. Navajas 4 , I. Astigarraga 4,5 , A. Garcia-Orad 1,4 . 1 University of the Basque Country, Genetics, Leioa, Spain; 2 University Hospital Donostia, Pediatrics, San Sebastian, Spain; 3 University Hospital La Paz, Oncohematology, Madrid, Spain; 4 BioCruces Health Research Institute, Pediatrics, Barakaldo, Spain; 5 University Hospital Cruces, Pediatrics, Barakaldo, Spain Background: Methotrexate (MTX), key drug in childhood B-Acute Lymphoblastic Leukemia (ALL) therapy, often causes toxicity. Association between genetic variants in MTX transport genes and toxicity has been reported. It is known that these transporters are regulated by microRNAs (miRNAs). Despite miRNA SNPs interfere with miRNA levels or function, studies of miRNA polymorphisms and drug toxicity are almost absent. Regarding B-ALL, we have previously found rs56103835 in miR-323b associated with MTX plasma levels. Nowadays, a large amount of new miRNAs have been annotated. Therefore, the aim of this study was to determine if there are other variants in miRNAs associated with MTX levels. Methods: Blood samples of 167 Spanish patients with pediatric B-ALL treated with LAL/SHOP protocol were analyzed. We selected all the SNPs described in pre-miRNAs with a MAF >1% (213 SNPs in 206 miRNAs) that could regulate MTX transporters. Genotyping was performed with VeraCode GoldenGate platform. Results: Among the most significant results, we found rs59262801 in miR-5189, rs4909237 in miR-595 and rs78790512 in miR-6083 were associated with MTX plasma levels. These miRNAs were predicted, in silico, to regulate genes involved in MTX uptake: SLC46A1, SLC19A1 and SLCO1A2. Conclusion: In this study we detected 3 SNPs in miR-5189, miR-595 and miR-6083 that might affect SLC46A1, SLC19A1 and SLCO1A2 MTX transport genes regulation and could affect MTX levels in patients with pediatric B-ALL. This project was supported by RETICS (RD/12/0036/0060 and RD/12/0036/0036) and Basque Government (IT661−13). No conflict of interest.