ALBERT

Description: CTLA-4 is an inhibitory molecule that down-regulates T-cell activation. Although polymorphisms at CTLA-4 have been correlated with autoimmune diseases their association with clinical outcome after allogeneic hematopoietic stem cell transplantation (allo-HSCT) has yet to be explored. A total of 5 CTLA-4 single-nucleotide polymorphisms were genotyped on 536 HLA-identical sibling donors of allo-HSC transplants. Genotypes were tested for an association with patients' posttransplantation outcomes. The effect of the polymorphisms on cytotoxic T-lymphocyte antigen 4 (CTLA-4) mRNA and protein production were determined in 60 healthy control participants. We observed a reduction in the mRNA expression of the soluble CTLA-4 isoform in the presence of a G allele at CT60 and +49. Patients receiving stem cells from a donor with at least 1 G allele in position CT60 had worse overall survival (56.2% vs 69.8% at 5 years; P = .001; hazard ratio [HR], 3.80; 95% confidence interval [CI], 1.75-8.22), due to a higher risk of relapse (P = .049; HR, 1.71; 95% CI, 1.00-2.93). Acute graft-versus-host disease (aGVHD) was more frequent in patients receiving CT60 AA stem cells (P = .033; HR, 1.54; 95% CI, 1.03-2.29). This is the first study to report an association between polymorphisms at CTLA-4 and clinical outcome after allo-HSCT. The CT60 genotype influences relapse and aGVHD, probably due to its action on CTLA-4 alternative splicing.

Description: Abstract 3099 Background: Despite the fact that allogeneic SCT currently offers patients with high risk AML the best chance of cure, we've aimed to investigate the outcome of AML patients who have undergone ASCT in our center, considered as one of spanish reference hospital in SCT, and the parameters that have been able to influence in relapse rate (RR), overall survival (OS) and relapse free survival (RFS). Methods: Retrospective study in 121 AML patients who have undergone ASCT between 1988 and 2010. The analysis has been performed in 95 patients (50 male and 45 female) by excluding 26 acute promyelocitic leukemias (APL): 62 patients until 1999 and 33 since 2000. Median age was 45 [18–74] and median lecocyte count, 17250/μL [1000–318000]. 90% were “de novo” AML and 92% were in first complete remission (1°CR). Cytogenetic risk was as follows: 64% intermediate, 29% high and 7% low. Conditioning regimens were oral BuCy (62%), CyTBI (24%) and intravenous BuCy (12%). Stem cell source was bone marrow (BM) in 46 patients (48%), but the use of peripheral blood (PB) was generalized since 1997. Colony-stimulating factors were used in 52% of total patients. Median time from last treatment was 94 days [30–285]. Results: The median follow-up of this study was 125 months [0–216]. OS at 1, 3 and 5 years were 59%, 46% and 44%. RFS at 1, 3 and 5 years were 60%, 49% and 48%. There was no relapse after 5 years from ASCT. Early mortality (before day +100) was 12% (9 in 11 patients between 1988 and 1999) and late mortality was 47% (34 in 45 patients because of relapse, with no significant difference between 1988–1999 and 2000–2010). Secondary malignancies incidence was 13% (5 in 6 patients suffered from haematologic neoplasms: 4 MDS at 43, 89, 90 and 97 months and 1 Follicular Lymphoma at 50 months), median age of these patients was slightly higher (53, 5 years old) and none of them had received TBI. Multivariable analysis showed that RFS at 5 years was influenced by: disease status at ASCT (55% if 1°CR vs 0% if ≥2°CR/PR/refractory disease, p

Description: Abstract 3010FN2 Background: There is a need to improve the conditioning regimens for allogeneic HSCT, both reducing the regimen related toxicity and improving the anti-leukemic effect. The myeloablative (MA) regimen consisting in intravenous (iv) busulfan (BU) with fludarabine (F) might be a better option than the conventional BU-CY (Cyclophosphamide) combination, since BU and F act synergistically against leukemia cell lines and previous studies with iv BU-F conditioning have reported an improved safety profile over BU-CY. Objective: We aimed to evaluate the efficacy and safety of the MA BU-F regimen, delivering iv BU in a one-daily dose after F infusion, since previous pharmacologic and clinical data supported its safety compared with the standard four-daily doses in the HLA identical sibling allogeneic HSCT setting. Patients: One hundred thirty seven consecutive adult patients with myeloid malignancies from nine Spanish institutions were recruited from 2005 to 2011. They had a median age of 47 years (range 19–74) and 60% were males. Diagnosis were AML: 80 (58.4%), AML secondary to MDS: 24 (17.5%), MDS: 23 (16.8%) and MPD: 10 (7.3%). At HSCT, the disease stage was 1st CR or untreated in 85 (62%) and more advanced stage in 52 (38%) cases. Patients having Karnofsky 〈 90% and Sorror CI 〉1 accounted for 11.7% and 38.3%. The conditioning regimen consisted in F, 40 mg/m2 daily for 4 days (total dose 160 mg/m2) followed by BU, one-daily IV 3.2 mg/kg infusion (total dose 12.8 mg/kg). GVHD prophylaxis consisted in cyclosporine and methotrexate. HSC infusion and post-transplant supportive measures and follow-up were made according each institution policies. Results: Donor graft source was peripheral blood (PB) in 93 (67.9%) and bone marrow (BM) in 44 (32.1%) cases. Median CD34+ cells infused were 4.5 millions/kg (range 0.6–17.8). All but one patient engrafted, with a median of 15 days (range 8–49) to 〉0.5 ×10E9 PMN/L and 13 days (range 7–149) to 〉20 ×10E9 platelets/L. The most frequent toxicity was mucositis, which was grade 〉1, in 68.8% cases. Three patients had hepatic SOS grade 〉1, and all of them resolved. Median hospitalization time was 30 days (range 17–114). Acute GVHD grade 2–4 incidence was 25% with a median of 32 days (range 12–87) to GVHD onset. The day-100 mortality was 4.8%. The chronic GVHD incidence in 114 patients at risk is 61.4%, with 37.7% of cases in extended form. At the time of this analysis, the median follow-up is 12 months (range 1–74). Crude survival data showed 106 (77.4%) patients remaining alive and 106 (77.4%) relapse free. Overall, 31 patients have died, 19 relapse-related (13.9%) and 12 (8.8%) due to transplant related mortality. Actuarial survival at 12 months is 79.4% and disease free survival is 71.3% in the whole series. Kaplan-Meier analysis showed that advanced age, comorbidities or advanced disease status at HSCT did not predict an inferior outcome. In conclusion, in the HLA identical sibling allogeneic HSCT setting, the BU-F regimen provides an adequate control of myeloid malignancies, with low regimen related toxicity and both reduced day-100 and non-relapse related mortality. Disclosures: Lahuerta: Janssen: Honoraria; Celgene: Honoraria.

Description: Abstract 4498 Background: Giving the fact that allo- SCT currently offers patients with high risk AML the best chance of cure, we`ve aimed to investigate the outcome of AML patients who have undergone allo-SCT in our center, considered as one of spanish reference hospital in SCT, and the parameters that have been able to influence in relapse rate (RR), overall survival (OS) and relapse free survival (RFS). Methods: Retrospective study in 192 AML patients who have undergone allo-SCT between 1982 and 2010. The analysis has been performed in 171 patients (85 male and 86 female) by excluding 21 acute promyelocitic leukemias (APL): 65 patients until 1999 and 106 since 2000. Median age was 37 1874 and median lecocyte count, atchmode documentclass[fleqn,10pt,legalpaper]{article} usepackage{amssymb} usepackage{amsfonts} usepackage{amsmath} pagestyle{empty} egin{document} (13400/hbox{ L }frac{470}{250000}) end{document}. 82 were de novo AML and 87 were in morfologic complete remission (70 in first CR). 14 patients had received a previous SCT. Cytogenetic risk was as follows: 55 intermediate, 34 high and 11 low. Conditioning regimen was ablative in 162 patients: CyTBI (36), BuCy (31), BuFlu (30) and others (3). 130 patients (76) underwent a related allo-SCT (95 of them were matched) and 41 patients (24), an unrelated allo-SCT (64 of them were matched). Stem cell source was bone marrow (BM) in 146 patients (85) and only 3 patients received umbilical cord (UC). Graft versus host disease (GvHD) prophilaxis was based on Ciclosporine in 150 patients (88). Median time from last treatment was 73 days 12268. Results: The median follow-up of this study was 61 months 1317. OS at 1, 3 and 5 years were 57, 44 and 40. RFS at 1, 3 and 5 years were 62, 50 and 45. Early mortality (before day 100) was 26 (43 until 1999 and 15 since 2000, p0,0001): 18 patients because of infections, 10 because of toxicity, 9 because of disease and 7 because of EICH. Late mortality was 27 (more than the half because of relapse, with no significant difference between 19881999 and 20002010). Cumulative relapse incidence at 5 years was 35, with a median time of relapse of 4 months. Secondary malignancies incidence was 5. Multivariable analysis showed that Transplantation Related Mortality (TRM) was influenced by: year of allo-SCT (OS at 5 years of 49 if 20002010 vs 28 if 19821999, p0,0001), late engraftment (p0,002) and severe acute GvHD (OS at 5 years of 45 if no evidence/grade I-II vs 25 if grade III-IV, p0,071). The other important parameters which lost its univariable analysis significance were donor type, recipient age and conditioning regimen. No difference was found in case of HLA and ABO discordance or donor/recipient CMV status. Multivariable analysis also showed that RR and RFS at 5 years was influenced by: disease status at allo-SCT (50 if 1CR vs 0 if 2CR/PR/refractory disease, p0,002), chronic GvHD (67 if present vs 41 if absent, p0,035) and leucocyte count at diagnosis (54 if 20000/ L vs 37 if 20000/ L, p0,038). The other important parameters which lost its univariable analysis significance were cytogenetic risk, initial induction response and positive minimal residual disease (MRD) before allo-SCT. No diference was found in case of ethiologic classification or stem cell source. Conclusions: Allo-SCT is a curative procedure in AML patients (global RFS of 50 at 3 years), specially when disease is under control and patient develops chronic GvHD. In the last decade, there have been important improvements in the procedure which have led to a significant decrease in TRM, and consequently, a significant increase in OS. Disclosures: No relevant conflicts of interest to declare.

Description: Background: Adults with high risk ALL features at diagnosis, slow responders or with recurrent disease have a poor outcome with standard chemotherapy and are considered for unrelated transplants in most centers if a matched sibling donor is not available. Unrelated cord blood (UCB) has emerged as an option for unrelated transplant in adult patients. Aim of the study: to compare the outcome of adult patients with unrelated transplant for poor prognosis ALL based on the hematopoietic source used for transplant. Patients and Methods: One hundred and forty- nine adult patients (median 29 years [15–59], 90M/59F) with poor prognosis ALL received an unrelated transplant in 13 Spanish institutions from 2000 to 2007. Patients in 1st CR at transplant met at least one adverse prognostic factor (adverse cytogenetics, hyperleukocytosis or a slow-responder). ALL was of precursor B lineage in 111 (74%), T-cell lineage in 28 (19%) and undetermined lineage in 10 (7%) patients. ALL was in 1st CR in 81 (54%) patients, in 2nd CR in 37 (25%), in 3rd CR in 11 (7%) and overt disease in 20 (13 %) patients. Both groups were comparable for the main clinical and biologic ALL features except for patients with more overt disease in UCB transplant group. Conditioning therapy consisted on TBI-CY in 68 (46%), BU-CY in 9 (6%), Thiotepa- BU-CY/FLU in 60 (40%) patients (all of them UCB transplants) and other regimens in 12 (8%) patients (6 of them were non-myeloablative). The source of hematopoietic progenitors was UCB in 62 (41%), mobilized PB in 41 (28%) and BM in 46 (31%) patients. HLA compatibility requirements for selecting unrelated donors (BM and mobilized PB) were 7–8 out of 8 allelic identities, and for UCB were 4–6 out of 6 A / B antigenic and DR allelic identities. Results: Acute GVHD was most frequent after PBSCT and BMT thanin UCBT. There was no significant difference for limited and extensive chronic GVHD between UCBT and PBSCT/BMT. The median follow-up was 16 months (0.3–101.4). DFS estimated at 5 years for patients transplanted with any source was not significantly better in transplants in 1st CR versus 2nd CR or more advanced disease (median 12 [2–22] vs 5 [2–9], p=0.106). There was no statistical difference in OS or DFS at 5 years between UCB and PBSCT/BMT). TRM was significantly lower in UCB transplants (p=0.021). The relapse probability was 17% for the PBSCT/BMT versus 29% for UCBT (p=0.088). The use of TBI as conditioning therapy was not associated with a lower relapse rate. 5y-OS (95%CI) Median OS (month, 95%CI) 5y-DFS (95%CI) Median DFS (month, 95%CI) 5y-TRM (95%CI) *p=0.021 Whole series 26% (17–35) 10 (4–17) 21% (13–29) 7 (3–11) 54% (44–64) PBSCT/BMT 22% (11–33) 9 (2–17) 21% (11–31) 5 (2–9) 63% (50–76)* UCBT 33% (18–48) 15 (7–22) 22% (8–36) 9 (1–17) 39% (25–53)* 1st CR ALL 30% (16–44) 12 (0–29) 24% (11–37) 12 (2–22) 57% (43–71) PBSCT/BMT 27% (9–45) 27 (2–51) 24% (8–40) 12 (2–22) 63% (44–82) UCBT 35% (14–56) 10 (0–23) 24% (4–44) 10 (0–26) 50% (31–69) GVHD was the main cause of death for unrelated PBSCT/BMT in contrast with infection and relapse for UCBT (p=0.001). Conclusions: UCB transplant and unrelated PBSCT/ BMT are equivalent options for poor prognosis adult ALL patients without a sibling donor. However, all modalities are associated with high transplant related mortality.

Description: Chronic Lymphocytic Leukemia (CLL) is heterogeneous and there are two clinical forms with different outcome, in which initial stage and parameters of tumor burden, expression of CD38 and /or ZAP 70, chromosome abnormalities such as 11q23 and 17p13 are associated with a more aggressive course of disease. The aim of this study is to analyse clinical and biological characteristics and outcome of patients with complex karyotype, and to asses if there are differences with other genomic aberrations. Fifty patients diagnosed of CLL between 1980 and 1994 were studied (median age: 71 yrs; female/male: 18/32; median follow up 6.6 yrs.). Biological studies with flow cytometry and fluorescence in situ hybridization were done on cryopreserved blood cells. Clinical stages at diagnosis were: Rai stage 0: 56%; I: 20%; II: 16%; III: 6%; IV: 2%. Binet A: 72%; B: 20%; C: 8%. At the time of diagnosis 37% of patients needed treatment and 61% presented disease progression. At the time of this study, 10% of patients are alive with a median follow up of 15.3 yrs. Seven patients (28%) presented complex karyotype. At the time of diagnosis 43% patients needed treatment and clinical stages were: Rai stage O: 29%, I: 14%, II: 43%, III: 14%, IV: 0%. Binet A: 43%; B: 43%; C: 14%. Median overall median survival was 72 months. The most frequent genomic aberrations associated with complex karyotype were: del (17p13) and trisomy 12 (60 %). Cellular expression of CD38 and ZAP 70 was not able to separate this group. Compared with other chromosome abnormalities, there were not important differences in clinical stage at diagnosis or expression of CD38 or ZAP 70, but overall median survival could distinguish three groups: del 11q22-23 or del 17p13 (34 months, p 〈 0,05) complex karyotype (72 months), and trisomy 12 or del 13q14 (79 months, p 〉 0,05). In conclusion, complex karyotype is frequently formed by del 17p13 and trisomy 12 and identifies a subgroup of patients with better prognosis compared with isolated p53 deletion. More studies have to be realized to determine if trisomy 12 is a cell defence mechanism.

Description: Expression of CD38 and /or ZAP 70 and chromosome abnormalities such as deletions of 11q23 and 17p13 have been previously identified by several groups as predictors of disease progression and decrease overall survival in Chronic Lymphocytic Leukemia (CLL). Moreover, few are described about the relationship between ZAP70/CD38 status and their genomic aberrations associated. We analysed 86 patients (median age: 70 yrs; female/male: 28/58) diagnosed of CLL between 1980 and 2005. Cytoplasmic ZAP-70 expression and cell surface expression of CD38, was determined by flow cytometry in CD19+ CD5+ B-CLL cells. Trisomy 12 and deletions in 13q14, 11q22–23 and 17p13, were detected by the interphase cytogenetic fluorescence in situ hybridization (FISH). Biological study in patients diagnosed before 2001 was done on cryopreserved blood cells. Concordant expression of ZAP-70 and CD38 was seen in 66 of 86 patients (76%). Fifty-one patients were ZAP-70- and CD38-, and 15 patients were ZAP-70+ and CD38+. Discordant ZAP70/CD38 status was present in 20 patients. Cytogenetic study identified chromosome aberrations in 85% of patients. Deletions in band 13q14 (43%), followed by trisomy 12 (21%), deletions in 11q22–23 (11%) and in 17p13 (10%) were the most frequent abnormalities. Normal karyotype was presented in 28% of patients and complex karyotype in 11%. In the concordant ZAP70-CD38- cases and ZAP70+ CD38+, 13q14 aberration (47, 9%) and normal karyotype (42%) respectively, were more frequent than in the other groups but in our study this differences were not statistically significant. Moreover deletion in 11q22–23 (25%) was seen statistically significant, p = 0, 05, in patients with concordant expression of ZAP70+ and CD38+. Patients with discordant status in the expression of ZAP70 and CD38, were statistically associated with trisomy 12 (38%) p = 0,002. In conclusion, in our study we find that expression in ZAP70 and CD38 is associated with a different genomic aberration. Deletion in 13q14 appears more frequent in patients with concordant ZAP70- and CD38- expression. Trisomy 12 is associated with discordant status expression. Finally, the worse prognostic group ZAP70+ and CD38+ is associated with a normal karyotype or deletions in 11q22–23. Expression of ZAP 70 and CD38 and genomic aberrations ZAP70+ CD38+ ZAP70- CD38- Discordant status p-value Del 11q22-23 25% 4,2% 12,5% 0,05 Del 17p13 8,3% 4,2% ---- 0,52 Trisomy12 8,3% 10,4% 37,5% 0,002 Del 13q14 16,7% 47,9% 31,3% 0,10 Normal 41,7% 33,3% 18,8% 0,39 Complex Karyotype 20% 5,9% 20% 0,13