Leukemia-free Survival In Patients Research Articles

Clinical Significance of Genetic and Molecular Changes in Primary Myeloid Sarcoma

To investigate the clinical significance of genetic and molecular changes in primary myeloid sarcoma (MS). Fourteen patients with primary MS were selected in Jiading District Central Hospital Affiliated Shanghai University of Medicine & Health Sciences, The First People's Hospital of Lianyungang from September 2010 to December 2021. AML1-ETO fusion, PML-RARα fusion and CBFβ breakage were detected by fluorescence in situ hybridization (FISH), and the mutations of NPM1, CEBPA, FLT3, RUNX1, ASXL1, KIT and TP53 genes were detected by new generation sequencing (NGS). Among 14 patients, the MS occurred in bone, breast, epididymis, lung, chest wall, cervix, small intestine, ovary, lymph nodes and central nervous system. The tumor cells expressed MPO (13 cases), CD34 (7 cases), CD43 (8 cases), CD68 (7 cases), CD99 (8 cases) and CD117 (6 cases). Cytogenetic abnormalities were observed in 4 cases, including 3 cases of AML1-ETO fusion and 1 case of CBFβ breakage, while no PML-RARα fusion was detected. There were no significant differences in overall survival (OS) and leukemia-free survival (LFS) between patients with and without AML1-ETO fusion/CBFβ breakage (both P >0.05). Among the 14 patients, the number of NPM1, CEBPA, FLT3-ITD, RUNX1, ASXL1, KIT and TP53 gene mutations was 5, 3, 5, 3, 2, 2, 1, respectively, of which 7 cases had at least one mutation in FLT3-ITD, RUNX1, ASXL1 and TP53 gene. The OS and LFS of patients with FLT3-ITD, RUNX1, ASXL1 or TP53 mutation were shorter than those without mutations (both P <0.01). The genetic and molecular abnormalities of primary MS can be detected by FISH and NGS techniques. FLT3-ITD, RUNX1, ASXL1 or TP53 mutation indicates a worse prognosis, but further clinical studies are needed to confirm it.

Validation of ELN 2022 Risk Classification in Patients Diagnosed with AML Undergoing Allogeneic Hematopoietic Cell Transplantation

Background: In 2022, the European LeukemiaNet (ELN) updated their guidelines (Döhner H et al., Blood 2022) for risk allocation of patients with acute myeloid leukemia (AML) based on essential genetic features at diagnosis as defined in the newest classifications (Arber D et al., Blood 2022, Khoury JD et al., Leukemia 2022). Methods: Retrospective analysis of the 147 consecutive patients diagnosed with AML and available genetic information who underwent alloHCT in a complete morphological response (CR) between 2011 and 2021 in a single institution (HCB). Data was collected retrospectively and updated in July 2023. G-banding karyotype, FISH, qualitative and quantitative PCR and targeted Next Generation Sequencing were assessed at diagnosis to classify patients according to ELN 2022 risk classification. Our primary goal was to validate the ELN 2022 risk classification in patients undergoing alloHCT analyzing overall survival (OS). Secondary objectives were analyzing leukemia-free survival (LFS), 24-month cumulative incidence of relapse (CIR) and in the different molecular subgroups within the ELN2022 risk groups. Results: The main patient, disease-related and transplant characteristics are displayed in Figure 1. Overall, the median age was 52 years (range 18-70), 53.7% of the patients males, with 88% of the patients undergoing alloHCT in CR1. Twenty-two patients could not be classified through NGS assessment between 2011 and 2016 and included in the intermediate risk category due to normal karyotype and wildtype recurrent abnormalities measured by PCR. Patients in the adverse-risk group were older (p &amp;lt;0.001) and underwent alloHCT less often with undetectable measurable residual disease (p = 0.006). Moreover, patients in the favorable or intermediate risk group underwent alloHCT more often in CR2 or CR3 (p = 0.02). In addition, alloHCT characteristics were similar for all the patients included. With a 67-month median follow-up for LFS (95CI: 55-76), the median OS and LFS in patients classified into the favorable and intermediate risk groups were not reached (5-year OS and LFS rate in favorable risk group: 81% (95CI: 73-89) and 72% (95CI: 63-81). 5-year OS and LFS rate in intermediate risk group: 62% (95CI: 56-68) and 64% (95CI: 58-70). Patients included in the adverse risk group had an OS and LFS of 23.7 months (15.3-NR, p &amp;lt;0.001 between intermediate and adverse risk. Figure 1B) and 15.2 months (5.79-NR, p=0.015) respectively, with a significant lower 5-year OS rate (40%, 95CI: 34-46), LFS rate (38%, 95CI: 32-44) and a higher CIR (18% in the favorable risk group (95%CI: 11-26) vs. 20% in the intermediate risk group (95%CI: 18-23) vs. 40% in the adverse risk group (95%CI: 38-44), p = 0.024). Based on these results, a subanalysis was conducted in the patients classified into the adverse group. Those patients diagnosed with complex karyotype, inv(3), t(3;3), del(17p) and/or TP53 mutated were reassessed into a new risk category group called: Adverse-Plus (AdvP).No different baseline characteristics between the AdvP patients and the other subgroups were found. AdvP patients presented a shorter OS (NR in the rest of subgroups vs. 8.73 months in AdvP, p &amp;lt; 0.001 for AdvP vs. adverse risk. Figure 1C), LFS (NR in the rest of subgroups vs. 5.2 months, p &amp;lt; 0.001), and a higher CIR (62.5% in the AdvP group, p &amp;lt;0.001 between adverse risk and AdvP). Interestingly, using this four-group classification including AdvP, the other risk groups presented similar OS, LFS, and CIR without statistical differences in between them Summary/Conclussion: ELN2022 risk classification discriminates properly accurate in patients undergoing alloHCT. Reclassifying AML adverse risk patients diagnosed with complex karyotype, inv(3), t(3;3), del(17p) and/or TP53 mutated in a different subgroup should be considered due to a much worse prognosis after alloHCT, although it remains mandatory in first complete response.

Clinical Characteristics and Survival Analysis of Patients with Chronic Myelomonocytic Leukemia

To investigate the clinical characteristics, prognostic factors and efficacy of hypomethylating agent (HMA) in patients with chronic myelomonocytic leukemia (CMML). The clinical data of 37 newly diagnosed patients with CMML was analyzed retrospectively, and their clinical characteristics and the efficacy of HMA were summarized. Kaplan-Meier and Log-rank test were used for univariate survival analysis, and Cox proportional hazards regression model was used for multivariate analysis. The median age at diagnosis was 67 years old. Their common manifestations included fatigue, bleeding, abnormal blood routine and fever. Most patients had splenomegaly. According to FAB classification, there were 6 cases of myelodysplastic CMML and 31 cases of myeloproliferative CMML, while according to WHO classification, 8 patients belonged to CMML-0, 9 patients to CMML-1 and 20 patients to CMML-2. At the time of diagnosis, the median white blood cell count was 32.84×109/L, median hemoglobin (Hb) was 101 g/L, median platelet count was 65×109/L, median absolute monocyte count was 9.53×109//L, median absolute neutrophil count (ANC) was 11.29×109//L and median lactate dehydrogenase (LDH) was 374 U/L. Cytogenetic abnormalities were found in 4 cases among the 31 patients who underwent karyotype analysis or fluorescence in situ hybridization detection. There were 12 patients who had analyzable results and gene mutations were identified in 11 cases, including ASXL1, NRAS, TET2, SRSF2 and RUNX1. Among the 6 patients who were treated with HMA and could be evaluated for efficacy, 2 patients achieved complete remission, 1 patient achieved partial remission and 2 patients achieved clinical benefit. Compared with the non-HMA treatment group, overall survival (OS) time was not significantly prolonged in the HMA treatment group. Univariate analysis showed that Hb<100 g/L, ANC≥12×109/L, LDH≥250 U/L and peripheral blood (PB) blasts ≥5% were significantly associated with poor OS, while WHO classification CMML-2, Hb<100 g/L, ANC≥12×109/L, LDH≥250 U/L and PB blasts≥5% were significantly associated with poor leukemia-free survival (LFS) (P<0.05). Multivariate analysis showed that ANC≥12×109/L and PB blasts≥5% were significantly associated with poor OS and LFS (P<0.05). CMML has high heterogeneity in clinical characteristics, genetic changes, prognosis and treatment response. HMA can not significantly improve the survival of CMML patients. ANC≥12×109/L and PB blasts≥5% are independent prognostic factors of OS and LFS in patients with CMML.

Next-generation sequencing revealed factors associated with cumulative incidence of relapse and leukemia-free survival in patients with newly diagnosed acute myeloid leukemia

BackgroundSeveral prognostic biomarkers have been validated for acute myeloid leukemia (AML), a heterogeneous hematopoietic malignancy. However, the factors associated with the cumulative incidence of relapse (CIR) and leukemia-free survival (LFS) in real-world patients with AML have not been well defined. MethodsThis study examined clinical and mutational data of 246 patients with newly diagnosed AML who received the traditional “3 ​+ ​7” regimen in PLA General Hospital from January 2008 to August 2020. Factors associated with CIR and LFS in patients newly diagnosed with AML were analyzed using next-generation sequencing. ResultsAdditional sex combs-like 1 (ASXL1) and Serine/arginine-rich splicing factor 2 (SRSF2) mutations were found to be associated with an increased risk of CIR and a reduced LFS in univariate analysis, while only SRSF2 mutations were associated with these factors in the multivariate analysis. Hyperleukocytosis maintained an independent effect on LFS in the multivariate analysis. Hematopoietic stem cell transplantation conferred a significant prognostic benefit on both CIR and LFS in our cohort. Furthermore, we validated the risk classification of patients with AML receiving traditional induction regimens across a broad age range. Based on next-generation sequencing results, we concluded that SRSF2 mutations were predictive of an increased risk of relapse, inferior LFS rates, and non-relapse mortality in patients with newly diagnosed AML. ConclusionThese findings indicate that patients with SRSF2 mutations might not benefit from the conventional “3 ​+ ​7” regimen. Our results may help in developing molecular stratification strategies and could guide treatment decisions for patients with newly diagnosed AML.

Efficacy and safety of CD19 CAR-T cell therapy for patients with B cell acute lymphoblastic leukemia involving extramedullary relapse.

To evaluate the efficacy and safety of CD19 chimeric antigen receptor (CAR) T cell therapy for patients with B cell acute lymphoblastic leukemia (B-ALL) involving extramedullary relapse. Fifteen patients with B-ALL involving extramedullary relapse who received CD19 CAR-T cell therapy in the First Affiliated Hospital, Zhejiang University School of Medicine from January 2016 to October 2021 were enrolled in this study. The overall survival and leukemia-free survival of patients were analyzed using Kaplan-Meier curves, and the response of extramedullary lesions in different locations following the CD19 CAR-T cell therapy was observed. Cytokine release syndrome (CRS), hematological toxicity, and immune effector cell-associated neurotoxicity syndrome (ICANS) during CD19 CAR-T cell therapy were analyzed. The median follow-up time was 7 (3-71) months, and 11 cases (73.3%) achieved complete response, median duration of complete response was 6 (2-27) months; 3 cases (20.0%) achieved partial response; 1 case (6.7%) got progressive disease. The overall response rate was 93.3% (14/15), and the overall survival rate was 80.0% (12/15) at the end of follow-up. The cumulative incidence of relapse was 40.0% (6/15) and relapse mortality rate was 20.0% (3/15). Until last follow-up date, 9 cases (60.0%) were still in disease-free survival. Among the 15 patients, 13 cases (86.7%) developed cytokine release syndrome (CRS) after cell infusion, including 7 cases with grade 1-2 CRS, 6 cases with grade 3 CRS; 1 case suffered from reversible ICANS; 15 cases (100.0%) developed B cell dysplasia; 12 cases (80.0%) developed severe hematologic adverse reactions; 2 cases (13.3%) had abnormal liver function; 1 case (6.7%) had abnormal renal function; 4 cases (26.7%) developed infection. The adverse reactions mentioned above were well controlled. CD19 CAR-T cell therapy shows explicit efficacy and controllable adverse reactions for B-ALL patients with extramedullary relapse.

HSPG2 overexpression independently predicts poor survival in patients with acute myeloid leukemia

Heparan sulfate proteoglycan 2 (HSPG2), also known as perlecan, is a large multi-domain extracellular matrix proteoglycan, which contributes to the invasion, metastasis and angiogenesis of solid tumor. However, very little is known about the effect of HSPG2 on acute myeloid leukemia (AML). This study aims to investigate the prognostic value of the HSPG2 gene in terms of overall survival and leukemia-free survival in patients with AML. Bone marrow mononuclear cells (BMMCs) from 4 AML patients and 3 healthy controls were processed for RNA-Sequencing (RNA-seq). The mRNA expression level of HSPG2 in BMMCs and CD34+ hematopoietic stem/progenitor cells (HSPC) obtained from enrolled participants and human leukemic cell lines was detected by RT-qPCR. Then the correlations between the expression of HSPG2 and a variety of important clinical parameters, such as median white blood cell (WBC) count and bone marrow (BM) blasts, were further analyzed. The expression level of HSPG2 was significantly upregulated in AML patients at the time of diagnosis, downregulated after complete remission and then elevated again at relapse. Moreover, HSPG2 expression was associated with median WBC count (P < 0.001), median hemoglobin (P = 0.02), median platelet count (P = 0.001), and BM blasts (P < 0.001) in AML patients. Patients with high HSPG2 expression had both worse overall survival (OS) (P = 0.001) and poorer leukemia-free survival (LFS) (P = 0.047). In the multivariate analysis model, HSPG2 was identified as an independent prognostic biomarker of AML. Taken together, these results indicate that HSPG2 overexpression was associated with poor prognosis in AML patients, and may be a prognostic biomarker and therapeutic target of AML.

Ikaros Family Zinc Finger 1 Mutation Is an Adverse Prognostic Factor for Patients with Adult B-Cell Acute Lymphoblastic Leukemia

To investigate the prognosis of patients with adult B-cell acute lymphoblastic leukemia (B-ALL) accompanied with Ikaros family zinc finger 1 (IKZF1) mutation, and to explore the role of allogeneic hematopoietic stem cell transplantation (allo-HSCT) in improving the clinical outcome of patients. The clinical data of 164 adult B-ALL patients who received IKZF-1 mutation detection by capillary electrophoresis of bone marrow were collected, and the relationship between the IKZF-1 gene mutation and the prognosis of adult B-ALL patients was analyzed. Among 164 adult B-ALL patients, the patients with IKZF-1 mutation (IKZF-1+) were 80 cases, while the patients without IKZF-1 mutation (IKZF-1-) were 84 cases. Among 80 IKZF-1 positive patients, according to the treatment method after complete remission these patients were divided into HSCT group (48 cases) and chemotherapy group (32 cases). Analysis showed that the 3-year overall survival (OS) and leukemia-free survival (LFS) rates in the IKZF1+ group were much lower. The OS and LFS rate in transplantation group were 50.3%±8.3%, 41.6%±8.5%; the OS and LFS rates in the chemotherapy group were 33.7%±12.8%, 31.5%±9.5%, which were lower than those in the IKZF1- group (the transplantation group: 79.5%±7.6%, 64.0%±8.4%; the chemotherapy group: 54.4%±9.9%, 40.6%±9.6% respectirely (P＜0.05). Among 80 B-ALL patients with IKZF-1- mutation, the 3-year OS and LFS rates were significantly higher in the transplantation group (55.3%±7.5%, 48.3%±7.6%) than those in the chemotherapy group (32.9%±11.8%, 28.4%±10.3%) with IKZF1 mutation (P＜0.05). IKZF1 mutation is an adverse prognostic factor for adult B-ALL patients, However, allo-HSCT significantly improves the OS and LFS of patients and also their clinical outcomes.

P53 protein overexpression in de novo acute myeloid leukemia patients with normal diploid karyotype correlates with FLT3 internal tandem duplication and worse relapse-free survival.

Although ~50% of acute myeloid leukemia (AML) patients have a normal diploid karyotype by conventional cytogenetics at diagnosis, this patient subset has a variable disease course and outcome. Aberrant overexpression of the p53 protein is usually associated with TP53 alterations and a complex karyotype, but the prevalence and impact of p53 overexpression in AML with diploid cytogenetics is unknown. We examined 100 newly diagnosed AML patients to evaluate the impact of p53 expression status quantified in bone marrow core biopsy samples using immunohistochemistry and computer-assisted image analysis. A total of 24 patients had p53 overexpression defined as 3+ staining intensity in ≥5% of cells; this finding correlated with lower platelet counts (P = .002), absence of CD34 expression in blasts (P = .009), higher bone marrow blast counts (P = .04), and a higher frequency of FLT3 internal tandem duplication (P = .007). Overexpression of p53 independently predicted for shorter leukemia-free survival in patients who underwent allogeneic stem cell transplantation by univariate (P = .021) and multivariate analyses (P = .004). There was no correlation between MDM2 and p53 protein expression in this cohort. We conclude that p53 expression evaluated by immunohistochemistry in bone marrow biopsy specimens at the time of AML diagnosis may indicate distinct clinical characteristics in patients with normal diploid cytogenetics and is a potentially valuable tool that can enhance risk-stratification.

Ikaros family zinc-finger 1 mutation is an independent factor for the poor prognosis of adult B-cell acute lymphoblastic leukemia, and allogeneic hematopoietic stem cell transplantation canimprove clinical outcomes.

To investigate the prognosis of patients with adult B-cell acute lymphoblastic leukemia (B-ALL) with Ikaros family zinc-finger 1 (IKZF1) mutation and determine the role of allogeneic hematopoietic stem cell transplantation (allo-HSCT) in improving the clinical outcome, we detected the IKZF1 mutation and BCR-ABL fusion gene at diagnosis in the bone marrow of 164 adult patients with B-ALL, and analyzed the clinical data of these patients retrospectively. Our analysis showed that grade III-IV acute graft-versus-host disease and IKZF1 mutation in the transplantation group and age and IKZF1 mutation in the non-transplantation group were independent factors for poor prognosis by univariate and multivariate analyses.The 3-year overall survival (OS) and leukemia-free survival (LFS) rates were much lower in the IKZF1+/BCR-ABL+ subgroup than in the IKZF1+/BCR-ABL- and IKZF1-/BCR-ABL- subgroups in both the transplantation and non-transplantation groups. The 3-year OS and LFS rates were significantly higher in the transplantation group than in the non-transplantation group with IKZF1 mutation.The study demonstrated that IKZF1 mutation was an independent factor indicating the poor prognosis of adult B-ALL and much worse prognosis in the BCR-ABL+ subgroup in both non-transplantation and transplantation groups. However, allo-HSCT significantly improved the OS and LFS of patients and also their clinical outcomes.

The predictors of leukemia free survival in patients with myelodysplastic syndrome

The predictors of leukemia free survival in patients with myelodysplastic syndrome

Prognostic Significance of Residual Acute Myeloid Leukemia in Bone Marrow Samples Taken Prior to Allogeneic Hematopoietic Cell Transplantation.

We sought to identify features in routine evaluation of pre-hematopoietic cell transplantation (HCT) bone marrow samples from patients with acute myeloid leukemia (AML) that influenced patient outcome. Of 140 patients, evidence of residual leukemia (RL) was identified in 38 (27%) of pre-HCT samples, as defined by 5% or more aspirate blasts, increased blood blasts, clustered or necrotic blasts on biopsy specimens, and/or leukemia-associated karyotypic abnormalities. Morphologic or karyotypic evidence of RL was significantly associated with shorter leukemia-free survival (LFS) compared with cases without identifiable RL (median, 7.1 vs 28.3 months; P < .0001). Upon multivariable analysis, RL, prior relapse, age, high-risk karyotype, and alternate donor source were each independently associated with shorter LFS. RL in pre-HCT samples was more strongly associated with shorter LFS in patients with intermediate or favorable-risk AML karyotype ( P = .001) than secondary or adverse karyotype-risk AML ( P = .04). Rigorous morphologic and karyotypic evaluation of pretransplant marrows is practical and important for posttransplant prognosis.

Treatment Outcomes and Prognostic Factors of Patients with Relapsed Acute Myeloid Leukemia after Allogeneic Hematopoietic Stem Cell Transplantation

Background: Allogeneic hematopoietic stem cell transplantation (HSCT) is the curative option for treatment of acute myeloid leukemia (AML). Even after HSCT, the relapse rate of AML ranged from 20% to 55%. The prognosis of relapsed AML after HSCT is very poor. But, treatment guideline for relapsed AML after HSCT has not been defined. Thus, we aimed to investigate treatment outcomes and prognostic factors of patients with relapsed AML after allogeneic HSCT.Patient and Method: Among 624 patients with AML who received HSCT in Asan Medical Center, Korea between 1995 and 2014, 219 patients who had a relapse of AML after HSCT were included in this retrospective study.Results: Seventy-two patients (32.9%) reattained complete remission (CR). Refractory AML at the time of HSCT (odds ratio [OR], 0.199, P=0.001), high BMI (> 28) (OR, 0.88, P=0.032), graft source from peripheral blood (OR, 0.299, P=0.001) and shorter relapse free interval (< 6 months) (OR, 0.199, P=0.001) were significant negative predictors for CR achievement (Table1). Of included patients, survival after relapse was median 5.2 months (95% CI, 3.96-6.44, months). Leukemia free survival of patients reattaining CR after relapse was median 10.6 months (95% CI, 8.1-13.2 months). Twenty-seven patients received second HSCT following salvage chemotherapy. Patients who had underwent upfront HSCT without chemotherapy for AML and relapse before 6months after HSCT were significantly associated with worse survival after relapse (Hazard ratio [HR], 3.106, P=0.006 and HR, 2.018, P=<0.001, respectively). Donor lymphocyte infusion following salvage chemotherapy and second HSCT had a significant impact on better survival (HR, 0.131, P=0.002 and HR, 0.423, P=0.003, respectively). Male gender (Hazard ratio [HR] 2.465, P=0.012) and extramedullary relapse (HR, 4.215, P=0.001) were negative factors affecting shorter leukemia free survival after relapse.Conclusion: Donor lymphocyte infusion following salvage chemotherapy and second HSCT can be beneficial option for relapsed AML patients after HSCT. But, novel treatment modalities need to be investigated for improving survivals of relapsed AML patients after HSCT. [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

Increased Number of Driver Mutations Is a Predictor of Response to Hypomethylating Agents in Patients with Myelodysplastic Syndromes

▪INTRODUCTION: Hypomethylating agents (HMA) such as azacitidine and decitabine remain the standard of care for the treatment of myelodysplastic syndromes (MDS). Although responses are seen in 40-60% patients, median response duration is 12-14 months. Eventual loss of response is associated with poor outcomes. Multiple studies have tried do identify biomarkers of response. Presence of TET2 mutations has been associated to increased response in several studies. Clear identification of predictors of response is still required.METHODS: We evaluated a total of 180 previously untreated patients with MDS or CMML that received HMA therapy at The University of Texas MD Anderson Cancer Center. Next generation sequencing (NGS), analyzing a panel of 28 genes (ABL1, ASXL1, BRAF, DNMT3A, EGFR, EZH2, FLT3, GATA1, GATA2, HRAS, IDH1, IDH2, IKZF2, JAK2, KIT, KRAS, MDM2, MLL, MPL, MYD88, NOTCH1, NPM1, NRAS, PTPN11, RUNX1, TET2, TP53 and WT1) was performed prior to therapy with HMA. Clinical and demographic data was obtained from clinical records. Generalized linear models were used to study association of response rates (ORR=overall and CR=complete) and risk factors. Response was defined following 2006 IWG criteria. The Kaplan-Meier produce limit methods were used to estimate the median overall survival (OS) and leukemia-free survival (LFS).RESULTS: Patient characteristics are shown in Table 1. Median age at diagnosis was 67 years (range 20-88). A total of 108 patients (60%) had lower-risk MDS based on IPSS and 72 (40%) had higher-risk MDS. A total of 143 patients (79%) had MDS and 37 (21%) had CMML. Eighty-seven (49%) patients had normal karyotype with 36 (20%) having complex karyotype. Therapy consisted in azacitidine monotherapy in 60 (33%) patients, decitabine monotherapy in 55 (31%) and guadecitabine or combinations in 65 (36%). The ORR was 58% (105/180) with 66 (37%) patients achieving CR. A total of 123 (68%) patients had at least one detectable mutation. Median number of mutations was 1 (range 0-5). The most frequently detected mutations included TET2 (23%), TP53 (16%) and RUNX1 (12%) which were all present in >10% cases. Frequency of detected mutations is shown in Figure 1A. Presence of ASXL1 mutation was associated with decreased likelihood of achieving CR (OR 0.30, 95% CI 0.10-0.93, p=0.037). No differences in ORR were observed based on presence of any individual mutation or any other clinical characteristic. Presence of TET2 mutation was not significantly associated with an increased likelihood of response (OR 1.32, 95% CI 0.64-2.70, p=0.453) or CR (OR 1.30, 95% CI 0.64-2.65, p=0.469). We subsequently analyzed the impact of number of detected mutations on response to therapy. Presence of three or more detectable mutations was associated with lower ORR (OR 0.29, 95% CI 0.10-0.88, p=0.028) and a trend to lower likelihood of achieving a CR (OR 0.22, 95% CI 0.05-1.01, p=0.052). Mutations in ASXL1 (p=0.003), BRAF (p=0.021), DNMT3A (p=0.018), EZH2 (p=0.01), GATA2 (p=0.021), MLL (p<0.001), NRAS (p<0.001), PTPN11 (p=0.002), RUNX1 (p=0.015), TET2 (p=0.001) and WT1 (p=0.04) tended to appear in patients with three or more mutations. Presence of TET2 mutations did not predict for ORR (OR 1.83, 95% CI 0.78-4.25, p=0.163) or CR (OR 1.75, 95% CI 0.80-3.81, p=0.159) within cases with less than three mutations. The median follow up of the cohort was 14.5 months (range 2.4-101.3 months). Patients who did not achieve a response had significantly shorter OS (median OS NR vs 21.3 months, HR 1.68, 95% CI 1.03-2.73, p=0.037) (Figure 1B) and LFS (median LFS NR vs 34.3 months, HR 2.13, 95% CI 1.00-4.50, p=0.049) (Figure 1C). Achievement of a CR predicted for improved OS in patients with higher-risk MDS (median OS NR vs 14.6 months, p=0.046) but not in lower-risk patients (NR vs 27.3 months, p=0.239). No significant differences in LFS were observed based on achievement of CR in both higher (p=0.238) and lower risk patients (p=0.453).CONCLUSION: The number of driver mutations may be a new biomarker to predict response to therapy with HMA in patients with MDS and CMML. Presence of ASXL1 mutations may be associated with a decreased risk of achieving a CR. As previously reported, response to therapy with HMA impacts the OS and LFS of patients with MDS. Incorporating sequencing data at diagnosis may help predict response to therapy and patient outcomes. [Display omitted] [Display omitted] DisclosuresJabbour:ARIAD: Consultancy, Research Funding; Pfizer: Consultancy, Research Funding; Novartis: Research Funding; BMS: Consultancy. Cortes:ARIAD: Consultancy, Research Funding; BMS: Consultancy, Research Funding; Novartis: Consultancy, Research Funding; Pfizer: Consultancy, Research Funding; Teva: Research Funding. DiNardo:Celgene: Research Funding; Daiichi Sankyo: Other: advisory board, Research Funding; Novartis: Other: advisory board, Research Funding; Abbvie: Research Funding; Agios: Other: advisory board, Research Funding. Daver:Sunesis: Consultancy, Research Funding; Ariad: Research Funding; Kiromic: Research Funding; Pfizer: Consultancy, Research Funding; Otsuka: Consultancy, Honoraria; BMS: Research Funding; Karyopharm: Honoraria, Research Funding. Konopleva:Cellectis: Research Funding; Calithera: Research Funding.

A Machine Learning Based Model to Predict Two-Year Leukemia Free Survival in Cord Blood Transplantation for Acute Leukemia - a Data Mining Study, on Behalf of Eurocord, Cord Blood Committee and the Acute Leukemia Working Party of the EBMT

Background: Umbilical cord blood transplantation (UCBT) is a potentially curative therapy acute leukemia (AL) patients. Transplantation benefit must be balanced against risks, such as transplant related mortality and relapse. The complex nature of hematopoietic stem cell transplantation data (HCT), rich in interactions and possibly nonlinear associations, has motivated us to apply machine learning (ML) for predictive modeling. ML is a field of artificial intelligence and is part of the data mining approach for data analysis.Our group has recently reported on a ML based prediction model for short term HCT outcomes (Shouval R et al; JCO 2015). Using a ML algorithm, the perspective of the current study was prediction of leukemia free survival (LFS) at 2 years after an UCBT, while exploring variables' importance and interactions.Patients & Methods: A cohort of 3,149 UCBT were analyzed. Inclusion criteria encompassed patients at all ages, undergoing an UCBT (single/double unit) in EBMT centers from the year 2004 to 2014, for AL, in all disease status. All conditioning and graft versus host disease prophylaxis regiments were included. A total of 24 variables were considered, including the number of total nucleated cell dose (TNC), donor and recipients HLA typing, as well as recipient, disease and transplant characteristics.The Random Survival Forest (RSF) ML algorithm was applied for model construction and data exploration. RSF is known to be adaptive to data, is able to automatically recover nonlinear effects and complex interactions among variables, and yields nonparametric prediction over test data. The analysis pipeline consisted of prediction model development, assessment of variable importance by their minimal depth from the tree trunk, and exploration of the top ranking variable with dependence plots. The latter promotes understanding of non-trivial associations between variables and outcomes.Results : The 2 years LFS was 49%, with a median follow up of 30 months. A RSF model of 1000 trees was developed, with each tree constructed on a bootstrap sample from the original cohort. A prediction error of 36.0% was calculated. The 10 most predictive variables (in ascending order) were disease status, age, TNC harvested and infused, recipient CMV serostatus, interval from diagnosis to UCBT, transplant year, previous autologous transplant, and use of anti-thymocyte globulin (ATG).Selected findings from exploration of variables-outcome relationship with dependence plots included a varying effect of TNCs in specific subpopulations. Increasing the number of infused TNCs had a positive effect on predicted LFS in patients receiving HLA mismatched (2 or more HLA mismatch) (figure) or single unit CB grafts, and patients in earlier disease status or older age. ATG administration was associated with worse LFS, whether unadjusted or adjusted to all other variables. However, there was an additional negative effect in advanced disease status patients, recipients of HLA mismatched or single CB units grafts, and older patients. Patients in 1st complete remission (CR) had higher predicted LFS as compared to those in 2nd CR. However, in patients receiving a HLA mismatched or a double CB graft, the difference in LFS between CR1 and CR2 was attenuated. Younger age had a favorable impact in early disease status, but lost its positive effect in advanced disease.Conclusions: A prediction model for LFS 2 years post UBCT was developed using the RSF ML algorithm. Variables were ranked according to their predictive contribution. Disease status, age, and TNC count were found to be the most important factors. Dependence plots revealed interactions and nonlinear associations between variables and the outcome, such as the effect of cell dose on HLA disparity. Apart from the study's clinical findings, it carries a methodological significance. A novel ML approach for prediction, variable selection and data exploration, accounting for long term time to event outcomes, has proved useful in the field of HCT. [Display omitted] DisclosuresMohty:Janssen: Honoraria; Celgene: Honoraria. Sanz:JANSSEN CILAG: Honoraria, Research Funding, Speakers Bureau. Bader:Neovii: Other: Institutional grants; Medac: Other: Institutional grants; Riemser: Other: Institutional grants; Amgen: Consultancy; Novartis: Consultancy; Jazz Pharmaceuticals: Consultancy.

In adults with t(8;21)AML, posttransplant RUNX1/RUNX1T1-based MRD monitoring, rather than c-KIT mutations, allows further risk stratification

AbstractWe asked whether minimal residual disease (MRD) determined by RUNX1/RUNX1T1 transcript levels could identify allogeneic hematopoietic stem cell transplantation (allo-HSCT) t(8;21) (q22;q22) acute myeloid leukemia patients who are at high risk for relapse, together with the impact of c-KIT mutations. Ninety-two consecutive adult t(8;21) patients who received allo-HSCT in complete remission were enrolled. MRD status at 1, 2, and 3 months after HSCT identified relapse patients (P = .05, P < .001, P = .0001, respectively). The 2-year cumulative incidence of relapse (CIR) and leukemia-free survival (LFS) was 32% vs 9% (P = .01) and 55% vs 70% (P = .12) for patients with and without c-KIT mutations, respectively. In multivariate analysis, MRD at the first 3 months after HSCT, rather than c-KIT mutations, was an independent factor for CIR (P = .001) and LFS (P = .001). In addition, 17 patients received donor lymphocyte infusion (DLI) as interventional therapy for MRD, and the 2-year CIR and LFS for patients with or without DLI was 24% vs 87% (P = .001) and 64% vs 0% (P < .001), respectively. In conclusion, MRD monitoring early after transplant allows further rapid identification of t(8;21) patients at high risk of relapse and was more predictive of relapse risk than c-KIT mutations.

Prognostic Value of Isocitrate Dehydrogenase Mutations in Myelodysplastic Syndromes: A Retrospective Cohort Study and Meta-Analysis

BackgroundRecent genomic sequencing efforts have identified a number of recurrent mutations in myelodysplastic syndromes (MDS) that may contribute to disease progression and overall survival, including mutations in isocitrate dehydrogenases 1 and 2 (IDH1 and IDH2).MethodsPretreatment bone marrow (BM) samples were acquired from mononuclear cells in 146 adult patients with de novo MDS from January 2006 to June 2013. Polymerase chain reaction (PCR) and direct sequencing were performed on exon 4 of IDH1/2 genes and mutation status was correlated with overall survival (OS) and leukemia-free survival (LFS). We then performed a meta-analysis combining previously published and current studies to explore the effect of IDH mutations on OS and LFS in MDS.ResultsIn our study, somatic mutations of either IDH gene were discovered in 11 MDS patients (7.53%) and were significantly correlated with poorer OS (P = 0.007). IDH mutations were specifically associated with a poorer OS in the intermediate-1 risk group by the International Prognostic Scoring System (IPSS) (P = 0.039). In addition, we discovered decitabine achieved a better therapeutic effect compared to other treatments in IDH mutation-positive patients (P = 0.023). We identified six previous studies of IDH mutations in MDS. A meta-analysis of these studies included 111 MDS patients IDH mutations and 1671 MDS patients with wild-type IDH1/2. The hazard ratios (HRs) of OS and LFS for patients with IDH mutations were 1.62 (95% CI, 1.27–2.09) and 2.21 (95% CI, 1.48–3.30), respectively.ConclusionThe results from our study and the meta-analysis provide firm evidence that IDH mutations are significantly associated with poorer clinical outcomes in MDS. Identification of IDH mutations may be pivotal for better risk stratification in MDS patients and improving IPSS score. Additionally, hypomethylating agents may be an effective treatment option for MDS patients with IDH mutations.

Clinical Significance Of SF3B1, U2AF1 and SRSF2 Spliceosomal Gene Mutations For The Treatment Of Hypomethylating Agents In Myelodysplastic Syndrome

BackgroundMany reports state that hematopoietic malignancies mostly result from somatic mutations in HSCs in the bone marrow. Somatic mutations of spliceosomal gene such as SF3B1, U2AF1 and SRSF2 have been widely described in myelodysplastic syndrome (MDS). Some studies presented that MDS patient with splicing factor mutations influence the clinical outcomes. However, the clinical significances for the treatment of hypomethylating agents (HMA) in splicing factor mutation were not reported. Therefore, this study investigated the influences of the SF3B1, U2AF1 and SRSF2 splice gene mutation in MDS patients who received the HMAs. Materials and MethodsMDS harboring ring sideroblast and association with somatic spliceosomal gene mutation was well demonstrated but, comparatively rare and showed good prognosis. So, we excluded MDS harboring ring sideroblast in this study. The study cohort of 133 MDS patients without harboring ring sideroblast was examined for somatic mutations in SF3B1, U2AF1 and SRSF2 splicing gene using direct sequencing method and 59 out of 133 patients received the treatment of HMAs (43 of Azacitidine and 16 of decitabine) for the treatment of MDS. Using the international prognostic scoring system(IPSS), the treatment indications for the HMA were as follows, 1) inermediate-1 with anemia and no response for the treatment of erythropoietin, 2) intermediate-1 with anemia accompanying other cytopenia ( neutrophil <1,000/uL or PLT <100,000/uL), 3) intermediate-2 or high risk. The response analysis was followed the modified IWG MDS response criteria. ResultsIn 59 patients, mutations in K700E of SF3B1; S34T, S34P or Q157P of U2AF1; P95H or P95R of SRSF2 were found in 6 (10.2%), 7 (11.8%), and 4 (6.8%) patients, respectively. The 17 patients were observed any mutation (SF3B1, U2AF1 or SRSF2) in 59 patients. We compared the clinical features, treatment responses and survivals according to the somatic mutations of spliceosomal gene vs wild type (WT) in each mutation. The disease composition of 59 patients was like as follows; 1 of MDS with del(5q), 6 of RCUD, 24 of RCMD, 9 of RAEB-1, 19 of RAEB-2. In the clinical features, lower risk (according to IPSS, WPSS and revised-IPSS) patients was included in the group with SF3B1 mutation (P<0.05). The hematologic improvement or more response for the HMA was observed in 33% vs 47% in SF3B1 mutation vs WT, 29% vs 48% in U2AF1 and 75% vs 44% in SRSF2, respectively. There was no difference in the response rates for the HMA therapy according to the mutation or wild type (P>0.05). Overall survival did not show the statistical differences in each mutation (P>0.05). The leukemia free survival in patients with SRSF2 mutation was inferior to the WT (p=0.001). However, anyone showed the leukemic transformation in the patients with SF3B1 mutation without statistical significance (p=0.247) (Fig. 1). [Display omitted] ConclusionOur results show that mutation of SF3B1, U2AF1 and SRSF2 spliceosomal gene in MDS patients without harboring ring sideroblast did not influence the treatment response and overall survival for the HMAs. However, alteration of SRSF2 splice gene may be regarded as a risk factor of leukemic transformation. So, the patients with SRSF2 mutation treated with HMA have to consider the aggressive therapy such as allogeneic stem cell transplantation before leukemic transformation. To confirm this result, it will be needed more study for large number of patients. Disclosures:No relevant conflicts of interest to declare.

Elevated C-reactive protein is associated with shortened leukemia-free survival in patients with myelofibrosis

Elevated C-reactive protein is associated with shortened leukemia-free survival in patients with myelofibrosis

Immunopharmacologic response of patients with B-lineage acute lymphoblastic leukemia to continuous infusion of T cell–engaging CD19/CD3-bispecific BiTE antibody blinatumomab

T cell-engaging CD19/CD3-bispecific BiTE Ab blinatumomab has shown an 80% complete molecular response rate and prolonged leukemia-free survival in patients with minimal residual B-lineage acute lymphoblastic leukemia (MRD(+) B-ALL). Here, we report that lymphocytes in all patients of a phase 2 study responded to continuous infusion of blinatumomab in a strikingly similar fashion. After start of infusion, B-cell counts dropped to < 1 B cell/μL within an average of 2 days and remained essentially undetectable for the entire treatment period. By contrast, T-cell counts in all patients declined to a nadir within < 1 day and recovered to baseline within a few days. T cells then expanded and on average more than doubled over baseline within 2-3 weeks under continued infusion of blinatumomab. A significant percentage of reappearing CD8(+) and CD4(+) T cells newly expressed activation marker CD69. Shortly after start of infusion, a transient release of cytokines dominated by IL-10, IL-6, and IFN-γ was observed, which no longer occurred on start of a second treatment cycle. The response of lymphocytes in leukemic patients to continuous infusion of blinatumomab helps to better understand the mode of action of this and other globally T cell-engaging Abs. The trial is registered with www.clinicaltrials.gov identifier NCT00560794.

Impact of Iron Overload in Higher Risk Myelodysplastic Syndromes

Abstract 2777 Background:Several studies demonstrated detrimental effect of iron overload and elevated serum ferritin mainly in lower risk myelodysplastic syndromes (MDS). Sanz and his colleagues reported inferior overall survival (OS) and leukemia free survival in patients with elevated serum ferritin ≥ 1000 ng/ml independent from RBC transfusion dependency (TD). Only 22% of the patients in that study were intermediate 2 (int-2) or high risk by international prognostic scoring system (IPSS). Recent studies also suggested inferior outcome with allogeneic stem cell transplant in patients with elevated serum ferritin. We examined the prognostic value of elevated serum ferritin in int-2 and high risk IPSS MDS patients. Methods:We identified int-2 or high risk IPSS MDS patients through Moffitt Cancer Center (MCC) MDS database. Patients were included if serum ferritin levels were available. Patients were divided into 2 groups based on serum ferritin <1000 or ≥ 1000 ng/ml. Individual charts were reviewed. Chi square test and t-test were used to compare baseline characteristics. Kaplan Meier estimates were used to calculate OS from time of referral to MCC, log rank test was used for comparison between the 2 groups and Cox regression analysis was used for multivariable analysis. All analysis was conducted using SPSS version 19.0 software. Results:We identified 201 MDS patients with int-2 or high risk IPSS evaluated at MCC. Serum ferritin data were available in 139 patients. Serum ferritin ≥ 1000 ng/ml was elevated in 38.3% (n=54). Baseline characteristics compared between the 2 groups included age, WHO subtype, MD Anderson risk model, karyotype, RBC TD, and treatment with azacitidine. No statistical significant differences were observed between the 2 groups except for a trend of more RBC TD in patients with elevated serum ferritin. (Table-1) The median duration of follow up was 34.9 months (95%CI 20.8–49.1). The median OS for patients with serum ferritin < 1000 ng/ml was 15.2 months (95%CI 12.6–17.9) compared to 8.1 months (95%CI 5.7–10.4) in patients with serum ferritin ≥ 1000 ng/ml (P=0.004).(Figure-1) In univariate analysis Age ≥ 60 years, MD Anderson risk group, RBC TD, serum ferritin and serum albumin were statistically significant prognostic factors for OS. In multivariable analysis, Age ≥ 60 years [HR 2.0 (95% CI 1.12–3.7), p=0.02], MD Anderson risk group [HR 1.8 (95%CI 1.3–2.54), p= <0.005], and serum ferritin ≥ 1000 ng/ml [HR 1.6 (95%CI 1.1–2.5), p=0.02] were independent prognostic factors for OS.The rate of AML transformation was 25.9% compared to 44.4% in patients with serum ferritin < 1000 and ≥ 1000 ng/ml respectively, p=0.039. This effect was not significant after adjusting for RBC TD.Table 1:Baseline characteristicsSerum ferritin <1000n=85Serum ferritin ≥ 1000N=54P valueAge<60 years16 (18.8%)9 (16.7%)0.75≥ 60 years69 (81.2%)45 (83.3%)WHO classificationRA11 (12.9%)7 (13.0%)0.36RARS8 (9.4%)2 (3.7%)RCMD10 (11.8%)11 (20.4%)RAEB56 (65.9%)34 (63%)KaryotypeGood24 (28.2%)12 (22.2%)0.86Intermediate15 (17.6%)11 (20.4%)Poor46 (54.1%)31 (57.5%)MD Anderson RiskLow2 (2.4%)2 (3.7%)0.90Int-112 (14.1%)6 (11.1%)Int-229 (34.1%)17 (31.5%)High42 (49.4%)29 (53.7%)RBC TDNo39 (46.4%)16 (30.2%)0.09Yes45 (53.6%)36 (67.9%)missing01 (1.9%)Azacitidine TreatmentNo34 (40%)25(46.3%)0.3Yes51 (60%)29 (53.7%) [Display omitted] Conclusions:Elevated serum ferritin level ≥ 1000 ng/ml in int-2 and high risk IPSS MDS was independently associated with worse OS and higher rate of AML progression. Those findings needs to be confirmed in larger cohort of patients and prospectively. Disclosures:No relevant conflicts of interest to declare.