ALBERT

Description: Abstract 3396 Introduction The risk of venous thromboembolic event (VTE) in patients with cancer is particularly increased compared to normal population. It seems that this risk depends largely on the characteristics of the tumor, such as its site or its stage of evolution, and the anti-neoplasic treatment. The capacity of thrombin generation and D-dimers levels are two biological markers proposed for the stratification of the risk of VTE. We have analyzed thrombin generation and D-dimers levels in patients with breast cancer as markers of a hypercoagulable state, depending on the stage and the period of the tumor evolution. Materials and methods It is a prospective study carried out at the day hospital of the carcinology department of the University Hospital H. Bourguiba in Sfax-Tunisia, including patients with breast cancer at different stages of evolution and different period elapsed since cancer diagnosis. At the time of inclusion, a venous citrated blood sampling (3.2%) was made. The test of thrombin generation was carried out according to the technique of Calibrated Automated Thrombogram assay (CAT®, Diagnostica Stago, Asnières, France). The parameters of the thrombogram were analyzed: endogenous thrombin potential (ETP), peak of thrombin (peak) and mean rate index (MRI). D-dimers were measured using the STA®- Liatest® D-Di assay (Diagnostica-Stago). Results Sixty one patients were included. Their average age is 51.8 ± 10.9 years old. Depending on the stage of cancer disease, 3 sub-groups of patients were distinguished: early local stage (T1, n=16), advanced local stage (T2-T4; n=25) and metastatic stage (M; n=20). Considering the time passed since diagnosis, we have different periods : inferior to 6 months (n=26), 6 to 12 months (n=7), 12 to 36 months (n=15) and more than 36 months (n=13). The analysis of the different parameters of the thrombogram depending on the cancer stage revealed that patients with an advanced local stage and a period elapsed since cancer diagnosis inferior to 6 months had significantly higher values of ETP and thrombin peak (1708±247 nM.min and 379±80 nM) compared to those with an older cancer (1404±308 nM.min, p

Description: In 105 consecutive patients with de novo acute myeloid leukemia (French-American-British M3 excluded), we compared prospectively the risk of bleeding complications, the number of platelet and red blood cell transfusions administered, and the costs of transfusions using two different prophylactic platelet transfusion protocols. Two hundred sixteen cycles of induction or consolidation chemotherapy and 3,843 days of thrombocytopenia less than 25 × 109/L were evaluated. At the start of the study, each of the 17 participating centers decided whether they would use a 10 × 109/L prophylactic platelet transfusion trigger (group A/8 centers) or a 20 × 109/L trigger (group B/9 centers). Bleeding complications (World Health Organization grade 2-4) during treatment cycles were comparable in the two groups: 20 of 110 (18%) in group A and 18 of 106 (17%) in group B (P = .8). Serious bleeding events (grade 3-4) were generally not related to the patient's platelet count but were the consequence of local lesions and plasma coagulation factor deficiencies due to sepsis. Eighty-six percent of the serious bleeding episodes occurred during induction chemotherapy. No patient died of a bleeding complication. There were no significant differences in the number of red blood cell transfusions administered between the two groups, but there were significant differences in the number of platelet transfusions administered per treatment cycle: pooled random donor platelet concentrates averaged 15.4 versus 25.4 (P 

Description: Genomic profiling in AML has led to increased understanding of oncogenic mutations, refined risk stratification, and enhanced identification of alterations that can serve as targets for therapeutic intervention. Comprehensive genomic profiling (CGP) identifies a variety of alterations, including base pair substitutions, insertions, deletions, copy number alterations, and fusions. As distinct age-dependent alterations in AML are being increasingly recognized, we sought to use CGP to gain insight into age-associate variants in pediatric and adult AML. Identification of the significant age-associated biologic differences is critical to improving understanding of the age-dependent drivers of leukemogenesis. This can also advance therapeutic efforts with enhanced risk stratification, disease monitoring, and drug development. Diagnostic and relapse specimens from a total of 934 patients, comprised of 179 pediatric (age 0-18 years) and 755 adult (age 19-87 years) specimens underwent clinical comprehensive sequencing. DNA and RNA integrated next-generation sequencing was performed in a CLIA-certified, CAP-accredited, NYS-approved laboratory for FoundationOne Heme. All captured libraries were sequenced to high depth averaging 569X for DNA (405 genes) and 〉3M unique pairs for RNA 9265 genes). Somatic variants identified included short variants (single nucleotides variants (SNVs) and short insertions, indels), and structural variants (fusions, amplifications, loss of whole genes, or truncations). The total variant prevalence was 571 in 131 genes in the pediatric cohort vs. 3020 variants in 219 genes in the adult cohort. The average number of variants in the pediatric cohort was 3.2 vs. 4.0 in the adults (p

Description: Childhood AML is a heterogeneous hematologic disease with a multitude of subtypes characterized by varying morphology, structural alterations, and recurrent mutations. Such heterogeneity and staggering number of genomic and transcriptional alterations has precluded appropriate risk classification. We investigated the expression of long non-coding RNAs (lncRNA) in childhood AML and explored its potential utility for more precise risk characterization at diagnosis. We inquired whether lncRNAs aberrantly expressed in AML compared to healthy normal bone marrows may be utilized for predicting outcome without knowledge of somatic variants, creating a variant-agnostic prognostic indicator. Ribodepleted RNA-sequencing data from normal bone marrows (N=68), as well as diagnostic primary samples (N=1300) from patients with detailed clinical annotations and outcome were included for study. To this end, the study population was divided into training (N=781), test (N=261), and validation (N=258) cohorts using blocked randomization. Upregulated lncRNAs compared to normal marrows in the training set (fold-change 〉 2, FDR 〈 0.05, Fig. 1A) were input for a LASSO cox proportional hazards regression which identified a set of 37 lncRNAs whose expression (log2 scale, TMM normalized) associated with patient outcome. The trained model coefficients were applied to the test and validation cohorts to produce a weighted sum of the 37 lncRNAs expression per patient (lncScores, range: -1.44 to +1.41). The distribution of lncScores revealed approximately equal numbers of patients with positive and negative scores in the training, test, and validation cohorts (Fig. 1B). In the training set, those with positive lncScores had an overall survival (OS) of 42.7% at 5-years from diagnosis compared to 76.3% for those with negative scores (hazard ratio (HR) = 3.15, p 〈 0.001). Positive lncScores were also associated with adverse event-free survival (EFS, HR = 2.47, p 〈 0.001). LncScore based outcome analysis in the independent test and validation cohorts showed nearly identical outcome results for OS (HR = 2.86 and 2.99 resp., p 〈 0.001) and EFS (HR = 2.37 and 2.35, p 〈 0.001, Fig. 1C) as was seen in the training set, demonstrating stability of the predictive power of the lncScore across independent cohorts. Next, the lncScore's performance was evaluated in relation to cyto/molecular risk groups (CMrisk: high, standard, and low) defined by karyotype, clinically relevant mutations, and cryptic fusions as presented previously (Cooper et al. ASH 2017). A multivariable cox regression model (N = 1300) that included lncScore, CMrisk, and white blood cell count revealed that both lncScore and CMrisk groups remained independent prognostic factors for OS (p 〈 0.001) and EFS (p ≤ 0.001), suggesting the lncRNA signature may provide additional prognostic information to that defined by CMrisk status. Application of the lncScore to the CMrisk groups demonstrated that 173 of the 541 of patients classified as CMrisk high (32%) would be reallocated to the standard risk arm based on negative lncScores (OS 57.3% vs 31.8%, p 〈 0.001). Similarly, 40% of patients with CMrisk standard (132/327) would be reclassified as low risk by the lncScore system (OS 73.8% vs 52.4%, p 〈 0.001). lncScores did not revise risk status in CMrisk low patients. The ribbon plot (Fig. 1E) demonstrates reallocation of patients using the integrated risk stratification system. Integration of the CMrisk and lncScore demonstrated that in combination the two risk classification systems can provide a more precise assessment of risk status, since both CMrisk high and CMrisk standard cohorts can be further stratified by the lncScore (Fig. 1D). The integrated approach provided a comprehensive risk classifier that incorporates both cytogenetic data, as well as transcriptional evidence, which benefits more than 1 out of 5 patients (23%, 305/1300) with a more accurate determination of risk at the time of diagnosis (Fig. 1F). This study demonstrates development and validation (in two independent cohorts) of a lncRNA based, variant-independent prognostic signature (p 〈 0.001) that can effectively partition patients into high and low risk groups at diagnosis. It can also significantly enhance the predictive power of conventional cyto/molecular markers for more precise prediction of outcome prior to the start of therapy. Disclosures Farrar: Novartis: Research Funding.

Description: In 105 consecutive patients with de novo acute myeloid leukemia (French-American-British M3 excluded), we compared prospectively the risk of bleeding complications, the number of platelet and red blood cell transfusions administered, and the costs of transfusions using two different prophylactic platelet transfusion protocols. Two hundred sixteen cycles of induction or consolidation chemotherapy and 3,843 days of thrombocytopenia less than 25 × 109/L were evaluated. At the start of the study, each of the 17 participating centers decided whether they would use a 10 × 109/L prophylactic platelet transfusion trigger (group A/8 centers) or a 20 × 109/L trigger (group B/9 centers). Bleeding complications (World Health Organization grade 2-4) during treatment cycles were comparable in the two groups: 20 of 110 (18%) in group A and 18 of 106 (17%) in group B (P = .8). Serious bleeding events (grade 3-4) were generally not related to the patient's platelet count but were the consequence of local lesions and plasma coagulation factor deficiencies due to sepsis. Eighty-six percent of the serious bleeding episodes occurred during induction chemotherapy. No patient died of a bleeding complication. There were no significant differences in the number of red blood cell transfusions administered between the two groups, but there were significant differences in the number of platelet transfusions administered per treatment cycle: pooled random donor platelet concentrates averaged 15.4 versus 25.4 (P 

Description: The PIM family of serine/threonine kinases (PIM1, PIM2, PIM3) are important regulators of signal transduction that phosphorylate proteins essential for cell proliferation, survival, and apoptosis. The PIM kinases are constitutively active and broadly expressed in multiple tissues and up-regulated in various malignancies. We report the discovery of a novel fusion transcript encoding the kinase domain of PIM3 fused to SCO2, a cytochrome c oxidase assembly protein. In transcript sequencing (RNA Seq) of 68 pediatric AML cases, PIM3-SCO2 fusion transcript was computationally identified and experimentally verified in index cases and studied in an independent cohort of pediatric patients with AML. RNA-Seq performed on the Illumina HiSeq in 68 diagnostic specimens from children with AML treated on COG clinical trials. Sequence reads were mapped to human genome using Novoalign. Four computational methods including Defuse, TophatFusion, FusionMap, and Snowshoes-FTD, were utilized to identify fusion transcripts and after filtering to eliminate false positives, fusions were selected based on observation in 2 or more fusion methods and presence in chimerDB. PIM3-SCO2 was identified as an in-frame fusion transcript in 3 cases with Inv(16) and subsequently verified by RT-PCR and Sanger sequencing. Frequency validation was performed by semi-quantitative expression analysis of PIM3-SCO2 expression levels in 235 AML diagnostic specimens as well as 6 normal bone marrow (NBM) controls. PIM3-SCO2 fusion protein was assessed by Western blot on whole cell lysates from cases with the fusion transcript. After verification of the fusion, available whole genome sequencing data in matching cases was interrogated and failed to demonstrate genomic counterpart to this fusion transcript, suggesting that this fusion may be the result of transcriptional read-through; also called transcription-induced chimera (TIC). Such fusion transcripts are generated when genes in proximity on a genome strand are spliced together to generate a chimeric product. Frequency validation studies in 235 diagnostic specimens from COG AAML0531 demonstrated that PIM3-SCO2 fusion transcript was highly prevalent in AML and expressed in 187 of the 235 cases of AML (80%) with variable prevalence across different cytogenetic cohorts, with prevalence of 87% in CBF, 56% in MLL, 79% in normal karyotype, and 70% in those with “other” karyotypes (p

Description: Abstract 5133 Introduction Cancer is generally combined with an increased risk of venous thromboembolism. This risk is linked to a hypercoagulable state whose precise mechanism remains unclear. It depends on the histological type, the stage of cancer and the anti-neoplasic treatment. This study aims to identify a hypercoagulability in patients with breast cancer and to define the most relevant procoagulant markers. Materials and methods A prospective study was carried out at the day hospital in the cancer department of the University Hospital H. Bourguiba in Sfax-Tunisia, from January to June 2012, including only patients with breast cancer. The thrombin generation was carried out on a venous citrated sampling (3. 2%) according to the technique of Calibrated Automated Thrombogram assay (CAT®, Diagnostica Stago, Asnières, France). The parameters of the thrombogram were analyzed: endogenous thrombin potential (ETP), peak of thrombin (peak) and mean rate index (MRI). The rates of platelet-derived microparticles expressing phosphatidyl-serine (MPP-PS+) were determined by flow cytometry using monoclonal antibodies anti-CD41 and anti-annexin V marked respectively by phycoerythrin (PE) and fluorescein isothiocyanate (FITC). D-dimers were measured by the STA-Liatest D-DI (Diagnostica-Stago) technique. The procoagulant activity of phospholipids (PPL) was measured by STA®-Procoag-PPL (Diagnostica-Stago). The plasma levels of tissue factor (TFa) were measured in a one-stage kinetic chromogenic method. This home-assay measures the ability of TF-FVIIa to activate factor X. Results Sixty one patients were included. Their mean age was 51. 8 ± 10. 9 years old. The study of the thrombogram parameters showed a significant increase of the peak of thrombin (343. 8±79. 2 nM) and of the velocity or MRI (153±56 nM/min) in patients compared to control subjects (288±48 nM, p=0. 001 and 109±33 nM/min, p500 ng/ml). MPP-PS+ in cancer patients were significantly higher (9684±7865/μl) than in controls (695±361/μl, p

Description: Key Points We show feasibility of whole-exome sequencing on purified primary HRS cells and report recurrent genetic alterations characterizing cHL. B2M is the most frequently mutated gene in cHL, strongly associated with nodular sclerosis subtype, younger age, and better overall survival.

Description: Background: HL, independently of socio economic conditions, is often intrinsically more aggressive in developing countries, including in Tunisia (Tunis Med.1999 77:614). We evaluated clinical characteristics and outcome of HL pts included in a prospective multicenter Tunisian adult HL trial. Methods: Between 2002 and 2006, 251 consecutive eligible pts from 6 Tunisian departments with newly diagnosed HL were enrolled in a prospective trial (MDH 2002). Pts were staged using EORTC prognostic factors in early stages and the international prognostic scoring (IPS) in advanced stages. ABVD was used at 3 cycles for favorable (fav) early stages (G1), 6 cycles for unfavorable (unfav) early stages (G2) and stage IIIA (G3) and 8 cycles for fav advanced stages (IPS

Description: Allogeneic hematopoietic stem cell transplantation (allo-HSCT) is widely utilized as a curative treatment for malignant and non-malignant hematological conditions. Fluoroquinolone prophylaxis (FQ-P) is demonstrated to reduce the rate of blood stream infections (BSI) caused by gram-negative bacteria (GNB) during allo-HSCT and increases overall survival (OS), making this approach the standard of care. The available data show that during the transplantation period, the intestinal microbiome diversity profoundly decreases, which is associated with a significant increase in transplant-related mortality (TRM), acute graft-versus-host disease (aGVHD) related mortality and decrease in OS. FQ-P is reported to be a dominant factor in the perturbation of the gut microbiota, leading some centers to omit or modify transplant antibiotic prophylaxis regimens. The aim of the present study has been to evaluate the effects of FQ-P omission on the prevalence of gram-negative bacteria blood stream infections (GNB-BSI), GNB susceptibility to antibiotic treatment, mortality of patients with sepsis and overall TRM. This retrospective single-center study included all consecutive patients, admitted to the Rambam Department of Hematology for allo-HSCT between 01.01.2017 and 31.12.2019. The fact that at our center, FQ-P in allo-HSCT recipients was discontinued on 01.12.2018 allowed comparison of the outcomes in patients treated with and without such prophylaxis. GNB-BSI events registered within 30 days of admission were analyzed. The proportion of first-time GNB-BSI, the antibiotic susceptibility profile, day 30 and day 90 mortality among patients with GNB-BSI were compared. The assessment also included day 30 and day 90 overall TRM, mortality related to sepsis and aGVHD. During the evaluated period, 189 patients underwent allo-HSCT and were included in the analysis. FQ-P was administered to 125 patients and omitted in 64 individuals. GNB-BSI events occurred in 23 (18.4%) patients receiving FQ-P and in 17 (26.6%) patients who did not receive it (p=0.19). GNB susceptibility to FQ, piperacillin/tazobactam and meropenem increased from 38.1% to 58.8%, from 60% to 70.6% and from 85.7% to 94.1%, respectively, after FQ-P had been stopped (p=non-significant, NS). 30-day and 90-day mortality among patients with GNB-BSI did not increase in the post-FQ-P period (Table 1). Day 30 and day 90 overall TRM rates were 10.6% and 18.9%, respectively, with FQ-P versus 13.5% and 21.9%, respectively, without FQ-P (p=NS). Before FQ-P was stopped, sepsis was the cause of death in 56% of events and aGVHD in 16% and after FQ-P was stopped, the corresponding values were 46% and 23%, respectively (p=NS). FQ-P omission has not significantly increased the rate of GNB-BSI or affected the profile of GNB susceptibility to antibiotic treatment in patients undergoing allo-HSCT. Moreover, it has not significantly changed day 30 and day 90 mortality either among patients with GNB-BSI or in the entire study population. FQ-P omission in allo-HSCT recipients appears to be safe and its implementation could contribute to the preservation of intestinal microbiome diversity, potentially leading to improved post-transplant outcome. The findings of this study need to be further evaluated in large randomized trials. Disclosures No relevant conflicts of interest to declare.