ALBERT

Description: Abstract 4311 Objectives Oral mucositis is a common complication of high-dose chemotherapy and radiotherapy followed by hematopoietic stem-cell support (HSCT). We evaluated the efficacy and safety of calcium phosphate mouth rinse (Caphosol) for prevention and reduction of severity and duration of oral mucositis in patients treated with HSCT. Methods In 2009 23 patients were treated. Three received allogeneic stem cells (ALLO): 2- Hodgkin disease (HD), 1-myeloma multiplex (MM) and twenty autologous transplantation (AUTO): 8-non-Hodgkin lymphomas (NHL); 7- HD; 3-MM. For ALLO conditioning regimens were composed of: fludarabine 150mg/m 2 and melphalan 140mg/m 2. AUTO were treated with: BEAM: carmustine, etoposide, cytarabine and melphalan (HD and NHL) and melphalan 200mg/m 2 (MM). The source of hematopoietic stem cells was peripheral blood. Caphosol was prepared according to the manufacturer‘s instructions and administered 4 times daily, starting from the day before the beginning of chemotherapy till the end of hospitalization. Control group was composed of patients, who had been treated with HSCT previously, before “palifermin and caphosol era”. The groups were comparable according to number of patients, their age, type of disease, transplant and conditioning regimen. Oral mucositis was assessed with the use of the five-grade World Health Organization (WHO) oral-toxicity scale. Oral mucositis was evaluated daily after the beginning of transplantation procedure until discharge from the BMT Unit. The number of days with painkillers or antibiotics were estimated. Total parenteral nutrition was given according to the standards [Martin-Salces M, De Paz R, Canales MA et al (2008) Nutritional recommendations in hematopoietic stem cell transplantation. Nutrition 24: 769-775]. Treatment with antibiotics was started when neutropenic fever occurred. Safety was assessed on the basis of the incidence of adverse events. Statistical analysis was performed using Wilcoxon‘ test for analysis of differences between groups. Results Oral mucositis grade 2-4 was not observed. In patients treated with calcium phosphate mouth rinse oral mucositis grade 2-4 was not observed. Nobody had to receive opioid analgesics or total parenteral nutrition. 30% patients developed the first degree of oral mucositis 4-5 days' duration. In the control group OM was observed in all cases, 50% patients had III- IV degree. Median duration of OM was 10 and 12 days (range 5- 20) for auto- and allogeneic patients, respectively. As compared with control group, treatment with calcium phosphate mouth rinse was associated with significant reduction of the incidence of oral mucositis in II- IV degrees (0 percent versus 50 percent, p 〈 0.001), duration of oral mucositis (4/ 5 days vs. 10/12 days, p 〈 0.001), duration of pain-killers‘ treatment (0- 22 days vs. 0- 3 days, p 〈 0.001) and number of days with antibiotics‘ treatment (0- 7 days vs. 7- 20 days, p= 0.002). These differences were observed in both types of transplantation. No side effects of calcium/phosphate oral rinse were observed. Conclusion In comparison with control group, treatment with Caphosol was associated with the significant reduction of the incidence of oral mucositis in II- IV degrees, duration of oral mucositis, pain-killers‘ treatment and number of days with antibiotics‘ treatment. Calcium phosphate mouth rinse is a very promising medicine for prevention of oral mucositis for patients treated with high dose chemotherapy supported with hematopoietic stem cell transplantation. Disclosures: No relevant conflicts of interest to declare.

Description: Abstract 3269 Poster Board III-1 Allogeneic hematopoietic stem cell transplantation (alloHSCT) is a potentially curative treatment for patients (pts) with chronic myeloid leukemia (CML). AlloHSCT is associated with long-term disease-free survival in 40% to 80% pts transplanted in early chronic phase of disease. The probability of relapse for pts transplanted in first chronic phase is 10% to 20% at 5 years, and is even higher (30% – 60%) for pts who received transplant in advanced phases of CML. The significance of minimal residual disease (MRD) in this clinical setting is uncertain. We enrolled 63 consecutive pts with CML who had received an alloHSCT between 1995 and 2007 and had BCR-ABL transcript quantity measured by RQ-PCR method on at least 2 occasions during follow-up in the period starting 6 months after alloHSCT. The reverse transcription was preformed using SuperScriptIII and random hexamers. Quantification of BCR-ABL was performed by RQ-PCR assay according to ‘Europe Against Cancer' protocol. BCR-ABL expression was normalized with endogenous control ABL gene and expressed as a ratio BCR-ABL/ABL. According to the amount of BCR-ABL transcript detected in blood or bone marrow after alloHSCT pts were allocated into 3 categories, including pts with no-detectable or stable very low-level of BCR-ABL transcripts (ratio BCR-ABL/ABL below 0.005%), pts with fluctuating-low level of BCR-ABL transcripts (0.005 – 0.01%) and pts with high-level of BCR-ABL transcripts (0.01 – 0.1%). We didn't find any relationships between different BCR-ABL levels after alloHSCT and clinical parameters at the time of CML diagnosis or transplantation, including Sokal, Hasford and Gratwohl scores. Median time from alloHSCT to molecular relapse (MR) was 38 months (range, 8.5 – 88.5 months). The 3-year progression rate into cytogenetic or hematological relapse of CML since MR was 70%. This progression occurred at a median time of 1.4 months (range, 0 – 3.2 months). We found strong correlation between the levels of BCR-ABL transcripts after alloHSCT and a risk of relapse. The incidence of MR was 0%, 26%, 71% for the low-level, fluctuating-low level and high-level of BCR-ABL transcript (p

Description: Allogeneic hematopoietic stem cell transplantation (alloHSCT) in chronic phase of chronic myeloid leukemia (CML) is associated with long-term disease-free survival and potentially eradication of leukemic cells. The goal of early MRD detection is to allow timely therapeutic intervention before hematologic relapse. The concomitant detection of BCR-ABL mRNA following alloHSCT is strongly associated with relapse, though not absolutely predictive. The relapse risk decreases with increased time after alloHSCT. The detection of BCR-ABL mRNA is the most strongly associated with relapse shortly after HSCT but all patients need to be monitored indefinitely after transplantation by molecular techniques presumably at 3–6 months intervals. Real-time quantitative PCR (RQ-PCR) for BCR-ABL mRNA provides an accurate and reliable measure of response to therapy in CML. In this study we evaluated 412 available samples from 75 patients at 1 month to 10 years after allo HSCT. Quantification of BCR-ABL was performed by RQ-PCR assay according to the Europe Against Cancer (EAC) protocol. Peripheral blood/bone marrow samples were studied every 3–6 months after alloHSCT for the presence of BCR-ABL transcripts using RT-PCR/nested PCR and RQ-PCR. RT-PCR positive patients were analyzed further at monthly intervals. RNA isolation from mononuclear cells was performed by column method. Reverse transcription was performed using Super Script II and random hexamers. BCR-ABL level was normalized with control ABL gene and expressed as the ratio of BCR-ABL/ABL compared to diagnostic sample or median expression values of BCR-ABL/ABL from EAC protocol. In our group BCR-ABL/ABL ratio decreased at least 1000-fold in all patients after alloHSCT. RT-PCR became negative in 64.7% patients after first 90 days. In the group of 65 patients with RQ-PCR tests performed at least 1 year after alloHSCT, 12 (18.5%) patients were always negative (no BCR-ABL/ABL transcripts detected, at least 10−5 test sensitivity), 40 (61.5%) were persistently low-level positive (with the BCR-ABL/ABL ratio less than 0.02%) and 13 (20.0%) patients were stable, high-level positive with transcript levels exceeded 0.02% threshold. The molecular relapse was observed in three patients with 15–80 fold increased of BCR-ABL expression in the first year after SCT. Decreased immunosuppressive therapy allowed achieving molecular remission in two patients. One patient developed hematological relapse despite donor lymphocyte infusion (DLI). One patient developed cytogenetic relapse and was successfully treated with 400 mg imatinib daily dose and achieved molecular remission with a low-level BCR-ABL expression. Standard imatinib therapy was applied in seven patients prior to SCT without negative transplant outcome. We conclude that RQ-PCR is valuable method to quantitate BCR-ABL expression in CML patients after alloHSCT and allows monitoring the kinetics of BCR-ABL mRNA transcipts and is useful in prediction of the hematologic relapse.

Description: Abstract 2315 Introduction: Allogeneic hematopoietic stem cell transplantation (allo-HSCT) is the possible curative therapy for many hematological malignancies and other congenital or acquired disorders. Despite significant progress, this procedure is still associated with substantial morbidity and mortality. Various pretransplantation and transplantation-related clinical risk factors have been implicated, but so far there is no method to estimate the occurrence and severity of transplant-related complications. The most common complication after allo-HSCT which affects survival remains graft versus host disease (GvHD). Cytokines and chemokines are major mediators in the inflammation and immune responses, especially in alloreactivity, which plays major role during GvHD. Single nucleotide polymorphisms (SNPs) in genes coding cytokines and chemokines have been shown to influence GvHD and outcome after allo-HSCT. This study aimed to determine association between SNPs in the: CCL5/RANTES (CC-chemokine ligand 5/Regulated upon Activation, Normal T cell Expressed and Secreted) promoter gene at positions -28C/T (rs1800825) and -403C/T (rs2107538), and IL-2 at position -330 G/T (rs2069762), IL-10 at positions -1082 A/G (rs1800896) and -592 A/C (rs1800872), and TNFalfa at position -308 A/G (rs1800629) with the outcome of allo-HSCT performed between August 2003 and April 2009 in our center. Materials and methods: The allelic variants of these 6 SNPs were determined in 64 patient/donor pairs by real-time polymerase chain reaction (TaqMan-MGB). Patients characteristics were: median age 36 (range 18–65), underlaying diseases-hematologic malignancies in 62 and aplastic anemia – in 2 cases. Donors were HLA-identical sibling-in 46 and unrelated-in 18 transplants, the median age-37 (range 14–65). The studied end points were GvHD, oral mucositis, haemorrhagic cystitis, toxicity and venooclusive disease (VOD) of the liver and survival. Results: The allelic distribution of examined SNPs was similar to that reported in Caucasoid population. Acute-GvHD was recognized in 31/64 (48,4%) and chronic-in 34/64 (53%) of patients. Overall survival in analysed group was 56%. Our study revealed significant correlation between the occurrence and severity of aGvHD and the -403C/T SNP in patients (TT and CT vs CC genotypes-41,7% vs 31,4% respectively, P=0.025), IL-10-592 A/C in recipients (AA genotypes–80%, P=0.050) and IL-2–330 G/T in patients (GG and GT vs TT genotypes respectively-40,6% vs 28,1%, P=0.034) and their donors (GG and GT vs TT genotypes respectively-11% vs 3,6%, P=0.049). Correlation between aGvHD and differences in donors genotypes in the -403C/T SNP was indicative of the trend (P=0.095). Besides we found a significant correlation between liver toxicity after conditioning regimen and SNPs: TNFalfa -308 A/G in patients (GG vs AA and AG–50% vs 45%, P=0.043) and donors (GG vs AA and AG–52% vs 40%, P=0.032) and IL-10-1082 A/G (GG vs AA and AG–77,8% vs 43,6%, P=0.003). Moreover our results revealed trend toward correlation between the -403C/T SNP in recipients and development of VOD (TT and CT vs CC genotypes-30,8% vs 11,8%, P=0.070) and haemorrhagic cystitis (30,8% vs 17,6%, P=0.071). The only statistically significant correlation for -28C/T SNP was increased overall survival obtained by the Kaplan-Meier method for patients carrying CT genotype (100% vs 51,7% for TT-homozygotes, P=0.048). Besides we couldn't find any correlation between examined SNPs and the rest of analysed end points (P=ns). Conclusion: Our data revealed that SNPs in the CCL5/RANTES, IL-10, IL-2 and TNFalfa promoter genes are correlated with aGvHD, toxic complications and overall survival after allo-HSCT. These findings confirmed some previously published data, but many results remains variable between investigators, so should be studied prospectively and in the larger group of patients. Disclosures: No relevant conflicts of interest to declare.

Description: Abstract 1334 Background A number of molecular aberrations have been described in acute myeloid leukemia (AML). They disturb normal haematopoiesis and are response for leukaemogenic mechanism and drug resistance. Therefore, they seem to be of prime importance for treatment outcome. FMS-like tyrosine kinase 3- internal tandem duplication (FLT3-ITD) and mixed-lineage leukemia gene-partial tandem duplication (MLL-PTD) are leukemia associated aberrations which confer worse prognosis. Downstream regulatory targets of FLT3-ITD and MLL-PTD are not well defined. The over-expression of ABC transport proteins that function as a membrane pump responsible for the efflux of a range of chemotherapeutic drugs are known mechanism behind multidrug drug resistance (MDR) in AML. The p-glycoprotein (Pgp)-product of MDR1 gene and multidrug resistant protein-1 (MRP-1), breast cancer resistance protein (BCRP) encoded by the relevant genes negatively influence the results of AML therapy. Aims The aim of this study was to analyze the expression of the MDR-1, MRP-1, BCRP messenger RNA (mRNA) in relation to the response to induction chemotherapy and relapse and with pretreatment laboratory and clinical characteristics such as age (

Description: AIMS: Oral mucositis (OM) is a frequent complication of myeloablative therapy and hematopoietic stem cell transplantation (HSCT). Palifermin was found to reduce the incidence, duration and severity of OM induced by high-dose chemotherapy with HSCT (Bone Marrow Transplant.2007;40:983–8). However, additional data on the long-term safety of palifermin and its potential influence on graft versus host disease (GvHD) were missing. In this multi-center, non-randomized, matched-control study we assessed overall survival (OS), incidence and severity of acute/chronicGvHD (a/cGvHD) and incidence of secondary malignancies in patients with hematological diseases treated with HSCT who received palifermin as a prophylaxis for OM. METHODS AND RESULTS: One hundred and twenty patients with hematological diseases transplanted between December 2001 and December 2007 were enrolled to this study. Sixty patients (50%) received palifermin (60μg/kg/day) for three consecutive days before and after conditioning therapy (palifermin group). Median age of patients was 37.5 years (range, 19 to 63) in the palifermin group and 35.5 years (range, 18 to 64) in the control group. There were no statistical differences between studied groups in terms of other clinical characteristics, including gender, diagnosis, type of transplant, conditioning regimens or GvHD prophylaxis. Kaplan-Meier curves for OS were calculated and compared using the log-rank test. Fisher’s exact and the χ2 trend tests were applied for the analysis of the GvHD data. Twenty one (35%) autologous and thirty nine (65%) allogeneic HSCT (HLA-matched related and unrelated) were performed in each group. Following allogeneic HSCT, the incidence of aGvHD grade 1–4 was 28.2% and 38.4% (p=0.34) and cGvHD was 41% and 53.8% (p=0.70) in the palifermin and control groups, respectively. The incidence of aGvHD severity grade 0,I,II,III,IV was 69.2%, 5.1%, 17.9%, 2.5%, 2.5% in the palifermin and 61.5%, 12.8%, 12.8%, 10.2%, 5.2% in the control group, respectively (p〉0.4 for each). The incidence of limited and extensive cGvHD was 17.9% and 23% in the palifermin group versus 25.6% and 28.2% in the control group (p=0.32 and p=0.50, respectively). The estimated 2-years OS (72.8% and 79.8%, respectively) also didn’t differ significantly between studied groups (p=0.13). We didn’t observe any secondary malignancies in patients enrolled into the study. CONCLUSIONS: Administration of palifermin doesn’t seem to influence the incidence and severity of a/cGvHD, secondary malignancies occurrence and OS in patients with hematological diseases undergoing HSCT.

Description: BACKGROUND: Recent EBMT analysis showed that infections are responsible for 21% of deaths after allo-HCT and 11% after auto-HCT. However, the risk, types and outcome of infections vary between age groups. The aim of the study is the direct comparison of risk factors of incidence and outcome of infections in children and adults. PATIENTS AND METHODS: We analyzed risk factors for the incidence and outcome of bacterial, fungal, and viral infections in 650 children and 3200 adults who received HCT between 2012-2015. The risk factors were determined by multivariable logistic regression analysis. RESULTS: A total number of 395/650 (60.8%) children and 1122/3200 (35.0%) adults were diagnosed for microbiologically confirmed infection, including 345/499 (69.1%) and 679/1070 (63.5%), respectively after allo-HCT, and 50/151 (33.1%) and 443/2130 (20.8%) respectively, after auto-HCT. At 2 years after HCT, the incidences of microbiologically documented bacterial infection were 36.0% and 27.6%, (p