bluuebunny/crossref_metadata_2025 huggingface dataset:
Incidence of Thrombotic Complications with Use of Peg-Asparginase in Treatment for Acute Lymphoblastic Leukemia (ALL) in Adults and Young Adolescent Patients
Authors: Routledge David JM , Tomlins Joanna , Bloor Adrian , Mamat Kaz , Dennis Michael , Cavet Jim , Somervaille Tim , Sommerfeld Sven Armin , Kulkarni Samar
Published: November 2019

Introduction: Peg-Asparginase is routinely used in the chemotherapy regimens used for treatment of ALL. Incidence of thrombotic complications is well established in children but there is limited data in adult and young adolescent patients. This is retrospective analysis to assess the risk of thrombotic complications with use of Peg-Asparginase.

Methods and Results: 100 patients with Acute Lymphoblastic Leukaemia treated between 2007-2015 who received Peg-Asparginase containing chemotherapy regimen were evaluated for development of thrombotic complications and pancreatitis. There were 69 male patients and 31 female patients. Median age was 29 yr. (range: 16-68 yr). Route of administration for Peg-Asparginase was intravenous in 51 cases and intramuscular in 49 cases. Peg-Aspraginase was administered during induction phase and consolidation cycles according to the relevant chemotherapy protocols. The adverse events included deep vein thrombosis (DVT) in 15 (15%), cerebral venous thrombosis in 7 (7%), Pancreatitis in 2 (2%), DVT and pancreatitis in 1 (1%) and 1 patient had clinically suspected DVT (1%). Complication rate was similar with IV or IM route of administration. 10 patients in IM group (20.4%) and 17 patients in IV group (33.3%) had at least one complication with Peg-Asparginase (P=0.31). Risk of individual complications was similar in both groups (DVT: 14.3% vs. 21.6%, p=0.34; Pancreatitis 2% vs. 3.9%, p=0.57; Cerebral venous thrombosis 4%% vs. 9.8%, p=0.25). There was no difference in the incidence of thrombotic complications with gender, age at diagnosis and use of central venous lines.

Conclusions: There is a significant risk of thrombotic events including PE (18% risk of thrombotic events excluding CNS). Risk of CNS events is 7% in this population. Risk appears to be higher with IV administration but this is not statistically significant due to small sample size. Incidence of CNS events is similar to that reported in adolescents participating in UKALL2003 study. The analysis will be extended to evaluate other toxicities especially hepatic toxicity and identify if there are any high risk patients for CNS or thrombotic events including blood counts, clotting, and cytogenetics. Use of LMWH prophylaxis seems reasonable in view of high incidence of thrombotic events even if CNS events are excluded.

Disclosures

Cavet: Celgene: Consultancy, Research Funding, Speakers Bureau; Janssen: Consultancy, Research Funding, Speakers Bureau. Somervaille:Novartis Pharmaceuticals Corporation: Consultancy, Membership on an entity's Board of Directors or advisory committees.
Two Cases of Pediatric Acute Lymphoblastic Leukemia with t(5;14): a Challenging and Unique Entity
Authors: Fortin-Boudreault Renee-Pier , Leung Elaine W. , Bassal Mylene
Published: November 2019

Background: Pre-B acute lymphoblastic leukemia (B-ALL) with the t(5;14) translocation occurs in less than 1% of B-ALL diagnoses. This translocation links the IGH on chromosome 14 with IL-3, which results in hypereosinophilia. Interestingly, circulating blasts or cytopenias usually don't accompany the eosinophilia. Patients often present with symptoms related to the eosinophilia, which can be quite morbid, and can delay the diagnosis of ALL. Few pediatric patients with this association have been described in the literature. The presentation and outcome can be quite variable, although it is thought that the prognosis in those patients is worse than standard ALL.

Objectives: To review the laboratory and clinical features of pediatric ALL with t(5;14).

Methods: Cases of pediatric patients with ALL and t(5;14) diagnosed at the Children's Hospital of Eastern Ontario between 1995 and 2003 were reviewed.

Results: We present two 11 year old children, a female and a male, who were diagnosed with B-ALL with t(5;14). The first one presented with a persisting low-grade fever and weight loss as well as asymptomatic lung infiltrates on the chest xray. The second patient presented with chest pain, fever, abdominal pain, as well as a petechial rash and splinter hemorrhages. His EKG showed ST depression, his troponins were elevated and an echocardiogram showed heart dysfunction. He went on and developed behavior changes and cerebral microinfarcts. Common to their presentation was the presence of hyperoesinophilia and absence of circulating blasts. Both had aggressive disease with persistent positive minimal residual disease (MRD) after consolidation and reinduction, which sent them to stem cell transplant.

Conclusion: ALL with t(5;14) is a rare entity that usually presents with hypereosinophilia. While eosinophilia can be asymptomatic, it can also be the cause of important morbidity. Diagnosis can be delayed because of the absence of blasts in the peripheral blood and lack of severe cytopenia. Finally, due to its rarity, there is very little information available on how this cytogenic abnormality impacts prognosis, which seems to be worse than ALL without this translocation.

Disclosures

No relevant conflicts of interest to declare.
Prevalence and Characteristics of Survivors from Adult Acute Myeloid Leukemia (AML) in Sweden 2014
Authors: Juliusson Gunnar , Lazarevic Vladimir , Antunovic Petar , Derolf Asa Rangert , Garelius Hege , Lehmann Sören , Myhr-Eriksson Kristina , Möllgård Lars , Uggla Bertil , Wahlin Anders , Wennström Lovisa , Höglund Martin
Published: November 2019

The prevalence of survivors of adult AML has previously been calculated, but the prevalent population has not been characterized. We analysed all 6289 patients diagnosed with adult AML from 1997-2013 as reported in the Swedish AML registry, with a complete survival follow-up. We found 1148 patients, 563 males and 585 females, who were alive on January 1, 2014, constituting 18% of the total, and of them 204 (18%) were diagnosed during 2013. Patients who had their initial treatment at pediatric departments, and people diagnosed with AML before 1997 are not included. The mean age of survivors was 53.4 years at diagnosis and 59.3 years in 2014, and 303 (26%) were 70 years or older in 2014. The overall prevalence of patients diagnosed at age 20 or more was 15.2 per 100,000 (males 15.0 and females 15.4 per 100,000). There were 124 with prior acute promyelocytic leukemia (APL) (11%), 54 with inv(16) (5%), 44 with t(8;21) (4%), and 82 with NPM1 mut/FLT3 wt (7%), 75 with FLT3 -ITD (7%), and 82 with complex karyotype (7%). Karyotype results were not available in 10%, and molecular data only since 2007. Four hundred two (35%) had undergone allogeneic stem cell transplantation, out of which 32 (8%) had had complex karyotype, 43 (11%) FLT3-ITD, 136 (34%) normal karyortype, and 41 (10%) good risk but were transplanted after relapse. Among the 426 non-transplanted prevalent patients who had survived at least 3 years, 94 (22%) had prior APL, 25 (6%) t(8;21), 24 (6%) inv(16), 126 (30%) normal karyotype, and 11 (3%) complex karyotype, including eight monosomal karyotypes and two with del(7) and one del(5q). Among non-complex karyotypes there were four MLL abnormalities, two del(5q) and two monosomy 7.

Although long-term survivors more often have pretreatment good-risk features, this patient group is still heterogeneous, and also contains older people and those with intermediate/high risk cyogenetics.

Figure 1. Prevalence in January 1, 2014 of Swedish people with a previous diagnosis of AML by sex and current age per 100,000 inhabitants in 2014, and mean incidence 1997-2014 per 100,000 inhabitants in Sweden 2005. Figure 1. Prevalence in January 1, 2014 of Swedish people with a previous diagnosis of AML by sex and current age per 100,000 inhabitants in 2014, and mean incidence 1997-2014 per 100,000 inhabitants in Sweden 2005.

Disclosures

No relevant conflicts of interest to declare.
Combination of Cladribine, Cytarabine and Topotecan (CLAT) for Relapsed or Refractory Acute Myeloid Leukemia
Authors: Chen Xiaomei , Weng Jianyu , Wang Yulian , Deng Chengxin , Luo Chengwei , Liao Pengjun , Li Minming , Huang Xin , Lai Peilong , Huang Lisi , Cao Zhi , Wan Changchun , Zhong Liye , Wu Suijing , Lu Zesheng , Zou Xiaoli , Du Xin
Published: November 2019

Xiaomei Chen and Jianyu Weng contributed equally to this study.

The outcomes ofrelapsed or refractory acute myeloid leukemia (RR-AML) are poor and effective salvage regimens are urgently needed. We present a study of 14 patients with RR-AML (median age 42years, range 18-65years; male n=12, female n= 2) treated with CLAT regimen, which consisted of cladribine 5mg/m² per day i.v. 2-3 hour on days 1-5, cytarabine 1.0g/m² per day i.v. 4 hours after cladribine on days 1-5, topotecan 1.25mg/m² per day i.v. 4 hours after cytarabine on days1-5 and G-CSF 300ug per day subcutaneous injection on days until neutrophile granulocyte recovery. Total of fourteen patients were included into the study from June 2013 to June 2015, Two (14.3%) patients were relapsed and twelve (85.7%) patients were refractory, 4 of 14 patients were relapsed or refractory after allogeneic-HSCT. Two patients died of invasive fungal infection before the assessment. Seven patients (58.3%) achieved complete remission (CR), and one patient (8.3%) achieved partial remission (PR), the rest patients (33.3%) did not respond (NR). The overall response rate was 66.7%. Following CLAT treatment, four patients with CR underwent allogeneic hematopoietic stem cell transplantation (HSCT) or microtransplantation. The median relapse-free survival (RFS) for RR-AML patients receiving CLAT regimen was 8.6 (range 2-16) months. Thirteen patients developed grade 4 granulocytopenia and thrombocytopenia, the median duration was 13(range 2 to 21) days and12 (range 2 to 21) days, respectively. The most common non-hematological side effects included nausea, vomiting, diarrhoea, and were grade 1/2. The CLAT regimen seems promising for the treatment of patients, and it was well tolerated. This regimen offers an alternative treatment for those patients with RR-AML who have received severe intensive treatment, especially with anthracycline-containing chemotherapy.

The project was sponsored by grants from National Natural Science Foundation of China (No. 30972790; No.81270648; No.81370665; No.81300446) Provincial Natural Science Foundation of Guangdong (No. S2012010009560) Provincial Science and Technology Planning Project of Guangdong (No.2013B021800186; No.2013B021800201), and Science and Technology Planning Project of Guangzhou (No. 201400000003-4, 201400000003-1).

Disclosures

No relevant conflicts of interest to declare.
Intensive Treatment Versus Azacitidine for Acute Myeloid Leukemia in Elderly: Retrospective Evaluation
Authors: Bolaman Zahit
Published: November 2019

Backround: Most of patients with acute myeloid leukaemia (AML) are elderly and older age is an negative prognostic factor for AML. There is no consensus about treatment of elderly patients with AML. In this study, we evaluated intensive treatment with cytosine arabinoside and idarubicine versus azacitidine treatment. Primary objectives were evaluation of disease free (DFS) and overall survival (OS), the rate of complete remission and the remission duration.

Methods: 45 patients with AML were evaluated retrospectively. We examined peripheral blood smear, bone marrow aspirate and trephine, immunophenotyping and cytogenetic analysis, biochemical evaluation of renal and liver function and coagulation profile in pre-treatment period. Intensive treatment consisted of cytosine arabinoside 200 mg/m2/day for 7 days and idarubicin 12 mg/m2/ for 3 days. Bone marrow examination was done between 14-28 days for intensive treatment and 2-6 cyclus for azacitidine treatment. Consolidation treatment (1-3) was done after intensive treatment if the patients were remission. All patients were ≤ECOG 2. Growth factors were not routinely administered before, during or after chemotherapy. Statistical analyses, differences in baseline characteristics between groups were analyzed using the Student's t-test for continuous variables and differences in OS between groups were then analyzed using the likelihood ratio or log-rank tests.

Table 1. The main properties of patients with elderly AML Treatment arm Mean age Sex M/F Leucocyte mean Hb gr/dl FAB (n) MDS history(n) LDH Complete remission Total 73±9 26/19 34.029 ±6870 8.53±1,2 M0:15, M1:6, M2:15,M4:4,M5:2 Yes:21 No:24 276±151 15 Intensive 72±7 18/10 44.176±12390 6.91±2 M0:6, M1:4, M2:12, M4:3, M5:1 yes: 14 299±101 13 Azacitidine 74±8 8/9 31.289±4013 8,95±1.6 M0:9, M1:2, M2:3, M4:1, M5:1 yes:7 256±72 2

Results: Total patient number were 45 and they were ≥ 65 years (yrs). Male/female ratio was 29/16. Two patients rejected leukemia treatment. One patient were not evaluated for treatment response. Intensive arm consisted of 26 patients and azacitidine arm 16 patients. Disease free survival and overall survival for intensive arm and azacitidine arm were 12,1±9,4, 16,7±22,4, respectively. The most importance factors were leucocyte counts, hb level.

Picture 1-2: Disease free survival and overall survival for intensive arm and azacitidine arm

Conclusions: These results showed that intensive treatment is more effective than azacitidine in elderly AML patients with ECOC≤2. There is need for large studies which include higher number of patients with AML in order to evaluate efficiency of intensive treatment.

Figure 1. Figure 1. Figure 2. Figure 2.

Disclosures

No relevant conflicts of interest to declare.
FLT3 Inhibitors for the Treatment of Acute Myeloid Leukemia: An Evaluation of Efficacy of Target Inhibition and Relationship to Disease Progression
Authors: Cloe Adam , Larson Richard A. , Cheng Jason X.
Published: November 2019

Introduction

FLT3 is a receptor tyrosine kinase that plays a role in hematopoietic stem/progenitor cell proliferation and survival and is frequently found to be mutated in patients with acute myeloid leukemia (AML). Mutations that lead to constitutive activation of FLT3 (such as internal tandem duplications of the juxtamembrane domain or point mutations involving the kinase domain) are associated with a poor prognosis. This poor prognosis is in part due to an increased relapse rate after allogeneic hematopoietic cell transplant (HCT). Several compounds that inhibit the activity of FLT3 in vitro, including ASP2215, midostaurin and quizartinib, are now being studied in clinical trials for the treatment of AML. Inhibition of the formation of phospho-FLT3 has been correlated with clinical anti-leukemia effects and remissions.

Methods

In this pilot study we use immunohistochemistry to measure the levels of activated FLT3 in bone marrow biopsies of patients prior to and during treatment inclinical trials at our institution to determine the efficacy of these FLT3 inhibitors and correlate it with patient outcomes.

Results

The different FLT3 compounds tested had a heterogenous effect on activated FLT3 levels. In some patients, ASP2215 caused a decrease in levels of activated FLT3 (indicated by brown nuclear staining), which corresponded to a decrease in leukemic blasts (Fig 1). Other patients, however, showed similar amounts of activated FLT3 both before and after treatment, which corresponded with no significant change in leukemic blasts. (Fig 2).

Other FLT3 inhibitors also showed differences in their effects. Midostaurin reduced activated FLT3 levels, which correlated with a positive clinical response. In contrast, one patient receiving quizartinibshowed little to no decrease in activated FLT3 levels, despite remaining in clinical remission. This suggests that this FLT3 inhibitor may have alternative targets, such as the tyrosine kinases AXL or LTK.

Conclusions

The heterogeneity in the responses to ASP2215, midostaurin, and quizartinib suggests that there may be other targets for these compounds that are not currently accounted for in the clinical studies. Immunohistochemicalmeasurements of activated FLT3 in bone marrow sections before and after treatment with FLT3 inhibitors was not predictive for clinical response.

Figure 1. Activated FLT3 levels in patient responsive to ASP2215; pre-treatment (A) and post-treatment (B) Figure 1. Activated FLT3 levels in patient responsive to ASP2215; pre-treatment (A) and post-treatment (B) Figure 2. Activated FLT3 levels in patient not responsive to ASP2215; pre-treatment (A) and post-treatment (B) Figure 2. Activated FLT3 levels in patient not responsive to ASP2215; pre-treatment (A) and post-treatment (B)

Disclosures

Larson: Pfizer: Consultancy; Ariad: Consultancy, Research Funding; Novartis: Consultancy, Research Funding; Bristol-Myers Squibb: Consultancy.
Distinct Clinical and Cytogenetic Characteristics and Poor Prognosis in Children with Acute Erythroid Leukemia: A Report from the JPLSG AML-05 Study
Authors: Tomizawa Daisuke , Tawa Akio , Kiyokawa Nobutaka , Kinoshita Akitoshi , Miyachi Hayato , Yamashita Yuka , Horibe Keizo , Iwamoto Shotaro , Deguchi Takao , Shiba Norio , Moritake Hiroshi , Shimada Akira , Hasegawa Daisuke , Taga Takashi , Ishii Eiichi , Adachi Souichi
Published: November 2019

BACKGROUND: Acute erythroid leukemia (AEL) is a rare subtype of acute myeloid leukemia (AML) that represents less than 5% of AML in children. However, its clinical picture is not fully understood.

PATIENTS & METHODS: We conducted a retrospective analysis on AEL using data from the Japanese Pediatric Leukemia/Lymphoma Study Group (JPLSG) AML-05 study (registered at http://www.umin.ac.jp/ctr/index.htm as UMIN000000511) in which patients were enrolled between 2006 and 2010. Morphological, immunophenotypic, cytogenetic, and molecular diagnoses were confirmed by the integrated prospective central diagnostic system.

RESULTS: Among the 443 children (age 0 to 18 years old at diagnosis) with de novo AML (acute promyelocytic leukemia and Down syndrome patients are excluded), ten AEL patients (2.2%) were identified. All the cases were classified as M6 in French-American-British classification (eight as M6a, one as M6b, and one as M6). Notably, 8 were classified as AML with myelodysplasia-related changes (5 with morphological dysplasia and 3 with myelodysplasia-related cytogenetic abnormalities) and 2 as AEL in World Health Organization classification (ver.4). There were 6 males and 4 females. Median age and leukocyte count at diagnosis was 3.5 years old (range, 1-15 years old) and 4.7 x 109/L (1.1-16.1 x 109/L), respectively. Cytogenetically, 3 had complex karyotype, 2 had monosomy 7, and 5 had normal karyotype. Molecular analysis showed one case with NRAS mutation, one with both mutated KIT and WT1, and one with MLL-PTD. No case was FLT3 -ITD positive. Eight cases achieved complete remission after initial induction but 3-year event-free survival and overall survival rates were 30.0% (SE 14.4%) and 41.1% (SE 17.5%), respectively.

CONCLUSIONS: AEL has distinct features such as high frequency of myelodysplasia- related features and poor risk cytogenetics. Because the outcome seems poor, further genetic analysis is required for development of effective novel therapy.

Disclosures

No relevant conflicts of interest to declare.
The Expression and Mechanism of Tim3 and PD-1 in Patients with Acute Myeloid Leukemia
Authors: Li Caixia , Yu Xiao , Gu Caihong , Ma Chao , Liu Hong , Tong Wang , Zou Qiu , Ye Lu , Chen Xiaochen , Depei Wu
Published: November 2019

Objective: To investigatethe expression and the role of Tim3 and PD-1 on T cells of peripheral blood in patients with acute myeloid leukemia(AML). Consistently discuss the clinical significance of Tim3 and PD-1 co-expression on these cells and the mechanism of Tim3-mediated immune escape.

Methods: Collectedclinical data of 36 patients who had been diagnosed with AML by bone marrow MICM classification, and then gathered their heparin anticoagulation peripheral blood before any treatment. The heparin anticoagulation peripheral blood from 20 healthy volunteers was gathered as normal control. We used the methods of immune fluorescence labeling and flow cytometry to detect expression of Tim3 on T cells. Then detected co-expression of Tim3 and PD-1 on T cells, consistently with IFN-γ secreting level on T cells.

Results: 1.The proportions of CD4+ T cells and CD8+ T cells on lymphocytes in AML patients were (15.28±10.99)% and (9.19±7.54)% respectively, which were significantly lower than the proportion of normal controls [(31.12±2.22)% and (21.59±4.22)% respectively] (P<0.05). 2.The levels of Tim3 expression on CD4+ T cells and CD8+ T cells in AML patients were (4.77±3.560)% and (5.90±4.91)% respectively, which were significantly higher than the levels of normal controls [(0.73±0.62)% and (0.96±0.54)% respectively] (P<0.05). 3.Tim3 and PD-1 were co-expression on CD4+ T cells and CD8+ T cells in AML patients. 4.The IFN-γ secreting level in Tim3+ CD8+ T cells was significantly decreased than Tim3- CD8+ T cells (P<0.05).

Conclusion: 1.The high expression of Tim3 on peripheral blood T cells in AML patients mediate T cell exhaustion/dysfunction, which can be an important mechanism of immune escape in leukemia. 2.PD-1 and Tim3 co-expression On CD4+ T cells and CD8+ T cells in AML patients which were dificient in there ability to produce IFN-γ. Further investigations are needed to explore the correlation of co-expression PD1 and Tim3 with process of AML, thus probably making Tim3 become another new immunotherapy target.

Disclosures

No relevant conflicts of interest to declare.
IL-6 Released from BMSCs Suppresses Cytotoxic Effect of Ara-C and IL-6 Mediated Signaling Enhances Re-Growing of HEL Cells Following Ara-C Treatment
Authors: Park Hyejoo , Kim Daeyoon , Lee Chansu , Bae Eunkyung , Ahn Kwang-Sung , Koh Youngil , Lee Hyun Jung , Kim Hyo Jung , Lee Junglim , Yoon Hwi-Joong , Yoon Sung-Soo
Published: November 2019

Various factors released from BMSCs regulate the biologic behaviors of AML cells involving in responsiveness of chemotherapeutic treatment. Cytotoxic effect of Ara-C decreased when AML cells co-cultured with BMSCs. IL-6 expression was prominently increased when HEL cells were co-cultured with BMSCs. Even though IL-6 did not affect the growth of HEL cells, Ara-c mediated apoptosis was suppressed by BMSCs. Also, IL-6 induced the phosphorylation of AKT and its downstream gene (mTOR). Ara-C mediated H2AX mRNA was suppressed when Ara-C was treated with co-cultured HEL cells with BMSCs. Also, its expression was down-regulated in HEL cells co-treated with Ara-C plus 25 nM IL-6. Prevention of IL-6 mediated signaling by gp130 shDNA slightly suppressed Ara-C induced H2AX expression when gp130 shDNA transfected HEL cells were co-cultured with BMSCs under 10-6 M Ara-C treatment. In vivo model, we found that IL-6 expression levels in serum of mice detecting AML cells following Ara-C treatment were higher than in serum of mice not detecting residual AML cells. Even though somatic mutation of gp130 gene was not detected in the genome analysis of AML, the overall survival was statistically different depending on the IL-6 levels in serum of bone marrow. Our findings suggest that IL-6 releasing from BMSCs help AML cells to survive against Ara-C treatment resulting in developing relapase from enhancing the growth of minimal residual cells.

Figure 1. IL-6 suppresses Ara-C mediated apoptosis of HEL in vitro assay. Figure 1. IL-6 suppresses Ara-C mediated apoptosis of HEL in vitro assay. Figure 2. IL-6 suppresses Ara-C mediated H2AX expression in HEL cells. Figure 2. IL-6 suppresses Ara-C mediated H2AX expression in HEL cells. Figure 3. Survival curve depending on IL-6 level of AML patients bone marrow serum. Figure 3. Survival curve depending on IL-6 level of AML patients bone marrow serum.

Disclosures

No relevant conflicts of interest to declare.
Analysis of flt3 in Adult AML
Authors: Adnan Shady , Kandeel Eman Z , Ghareeb Mohammed , Eldessouki Ihab A.
Published: November 2019

FLT3 gene encodes a tyrosine kinase receptor, which is involved in proliferation and differentiation of hematopoietic stem cells. FLT3 mutations are frequently encountered in all AML subgroups specially in AML with t(15;17)- Acute Promyelocytic Leukemia (APL)-, and infers poor prognostic effect. FLT3-ITD mutations represent about 80% of FLT3 mutations. In this study, we aimed to assess frequency and correlation of FLT3-ITD mutation with different AML cytogenetic subgroups, also with MPAL cases and to study the prognostic role of this mutation with focusing on APL.

Between 2011 and 2014 in NCI, Cairo University, 212 newly diagnosed acute leukemia patients [198 AML (10 M0, 47 M1, 56 M2, 30 M3, 49 M4, 4 M5, 1 M6 and 1 M7) and 14 MPAL] were included in this study. Thorough Morphological, Immunophenotypic and cytogenetic analysis were done at initial presentation and fixed follow up time points (Day 14, 28, 36) with subsequent monitoring and follow up of patients. FLT3-ITD mutation was detected using RT-PCR.

FLT3-ITD mutation was detected in 44/198 AML patients (22.2%) and in 2/14 MPAL patients (14.3%). Correlation with cytogenetic subgroups show significant higher incidence in M3 patients 13/30 (43.3%, P=0.0371) in comparison to other types (20% M0, 20.6% M1, 20.1% M2, 13.4% in Monocytic Leukemia -M4 and M5-). In non-APL patients, favorable cytogenetic changes are more common in FLT3 wild group (19.7%)[27/137 (21 patients with t(8;21) and 6 with inv 16] than in FLT3 mutant group (6.5%) [2 with t(8;21) and none with inv 16]. There were insignificant difference between FLT3 wild and mutant group regarding achieving Compete Remission (CR) by Day 36 (P= 0.261), However Overall Survival (OS) of the wild group was high significantly better than that of mutant group (P=0.002). By the end of the 1st year only 10% of patient with mutant FLT3 were alive versus 30% of those with wild type, with 4 months median survival of mutant group versus 8 months of the wild group.

In APL patients, again there was no significance between both groups regarding achieving CR by Day 36 (P=0.06) and still OS of APL patients with wild FLT3 was significantly better than in those with mutant FLT3 (P=0.0182). By the end of 1st year, about 90% of patients with wild group are still alive while only 46% of the mutant group are alive with 2 months median survival.

We conclude that FLT3-ITD mutations infers poor prognostic effect even in the presence of favorable cytogenetic changes like t(15;17); which indicate that patient with such mutations should be assigned to aggressive therapy.

Disclosures

No relevant conflicts of interest to declare.
Identification of the Microrna Signature of Acute Myeloid Leukemia Patient s Regulatory T Lymphocytes
Authors: Rouas Redouane , Merimi Makram , Badram Bassam , Burny Arsene , Bron Dominique , Lewalle Philippe , Fayyad Kazan Hussein
Published: November 2019

There is a clear evidence of elevated numbers of CD4 regulatory T lymphocytes (Tregs) in solid and hematological malignancies in addition to possessing an increased suppressive function. This important feature has urged the scientific community to search for ways to interfere with Treg function during or after cancer therapy. But the underlying molecular mechanisms remain largely unknown. Thus, a special focus on microRNA regulation was the goal of our study.

In our previous study, we investigated the molecular profile of circulating Tregs in the peripheral blood (PB) of healthy volunteers and identified several miRs differentially expressed between CD4+CD25+CD127low (Tregs) and CD4+CD25-CD127low T cells using the TLDA technique.

In fact, a human Treg microRNA signature of AML patients at diagnosis has not been described yet. We thus investigated human CD4+CD25+CD127 low Tregs purified from the peripheral blood of AML patients and identified a signature composed of ten differentially expressed miRs (miR-18a, miR-101, miR-133a, miR-135a, miR-302c, miR-324, miR-511, miR-542, miR-618, and miR-758) in comparison to their negative counterpart. Among those miRs, six were upregulated (miR-18a, miR-101, miR-133a, miR-135a, miR-511, miR-618) whilst four were downregulated (miR-302c, miR-324, miR-542, miR-758). Many potential miRs potential targets were identified in the 3'UTR of genes indispensable for Treg function and are presently under investigation. Moreover, we are comparing Tregs from AML patients to those from healthy individuals in term of microRNA signature and biology.

Disclosures

No relevant conflicts of interest to declare.
Quantitative Detection of DNMT3A R882H Mutation in Course of Treatment of Acute Myeloid Leukemia Patients
Authors: Blau Olga , Berenstein Rimma , Baldus Claudia , Suckert Nikola , Rieger Kathrin , Nogai Axel , Pezzutto Antonio , Doerken Bernd , Blau Igor Wolfgang
Published: November 2019

Introduction: DNMT3A mutation is one of the most common somatic mutations in acute myeloid leukemia (AML) patients with normal karyotype. The most frequent mutation is located at R882 codon in the methyltransferase domain. Because of prognostic significance and high stability during the disease evolution, DNMT3A mutations might represent highly informative biomarkers for prognosis and outcome of disease.

Methods: Using allele-specific PCR with a Blocking reagent (ASB-PCR assay) for the quantitative detection of DNMT3A R882H mutation, we analyzed 350 follow-up samples from 28 AML patients in complete remission (CR) after induction and consolidation treatment and allogeneic stem cell transplantation (alloSCT). Seventeen patients included in the follow-up analysis harbored a NPM1 mutation. Using a well-established marker for the detection of minimal residual disease (MRD) allowed to estimate the stability of DNMT3A mutation in CR and complete molecular remission (molCR). In addition, we analyzed FLT3, IDH1, and IDH2 mutations in diagnostic and follow up samples and donor chimerism after alloSCT.

Results: We found the persistence of DNMT3A R882H mutations in all patients in CR after standard therapy. On the contrary, after alloSCT, DNMT3A R882H mutation was not found in patients with CR and complete donor chimerism. In relapse of leukemia, an increasing of both NPM1 and DNMT3A mutated alleles were shown all cases.

Conclusion: Persistence of DNMT3A mutation after standard chemotherapy could indicate the origin of mutation in the early pre-leukemic stem cells. The loss of correlation between NPM1 and DNMT3A in CR could be associated with leukemic clone evolution. It is impotant to note, that the removal of mutated leukemic stem cells after allo SCT indicates therapeutic options allo SCT for high risk AML patients. Increased of both DNMT3A and NPM1 mutated alleles in relapse indicates the presence of both mutations, at least partly, in the same leukemic clone.

We conclude that quantitative detection of DNMT3A R882H mutations at different time points of AML disease could provide additional information about the role of mutations in development and progression of AML.

Disclosures

Blau: BMS: Honoraria; MSD: Honoraria; Celgene: Honoraria, Research Funding; AMGEN: Honoraria; JAZZ Pharma: Honoraria; Shire: Honoraria; Janssen: Honoraria, Research Funding; Baxalta: Honoraria.
Functional Proteomics of Regulatory and Epigenetic Signaling in Normal and Malignant Hematopoiesis
Authors: Cifani Paolo , Shakiba Mojdeh , Brown Fiona , Dhabaria Avantika , Bauer Daniel E. , Kentsis Alex
Published: November 2019

Post-translational modifications of proteins are increasingly recognized as key regulators of functional complexes assembly and cellular enzymes activity, processes that control both normal and aberrant cell growth and development. To enable comprehensive, sensitive and quantitative analysis of these processes, we adapted recently developed high-efficiency nanoscale multidimensional liquid chromatography with high-resolution Orbitrap mass spectrometry. Termed accumulated ion monitoring (AIM), this mass spectrometry method achieved nearly 7 orders of magnitude of quantitative accuracy and absolute limit of detection of 600 molecules per scan, enabling the study of rare cell populations (Fig. 1). We leveraged AIM mass spectrometry to develop a panel of 1583 synthetic reference peptides, based on global and published proteomics maps of normal human and leukemia cells. This Quantitative Cell Proteomics Atlas (QCPA) profiles 384 key effectors of cell surface signaling, proliferation, quiescence, stress response, and epigenetic control of gene expression (Fig. 2). QCPA enables unprecedented accuracy and sensitivity for the functional analysis of rare cells, with focus on protein regulation via phosphorylation, acetylation and methylation, as well as on concentration (http://alexkentsis.net/qcpa). By using functional proteomics profiling of primary human CD34+ and acute myeloid leukemia (AML) cells, we identify new pathways controlling erythrocyte differentiation and AML therapy resistance, as facilitated by a newly developed program ProteoModlR (http://github.com/kentsisresearchgroup/ProteoModlR). AIM mass spectrometry and QCPA functional proteomics are rapidly adaptable and generalizable tools for the investigation of regulatory and epigenetic signaling in normal and diseased cells.

Figure 1. Limits of detection and quantitation for a serially diluted synthetic peptide from human Myocyte-specific Enhancer Factor 2C. Figure 1. Limits of detection and quantitation for a serially diluted synthetic peptide from human Myocyte-specific Enhancer Factor 2C. Figure 2. Functional classification of proteins included in the QCPA. Figure 2. Functional classification of proteins included in the QCPA.

Disclosures

Bauer: Editas Medicine: Consultancy; Biogen: Research Funding.
The Role of Interim F-18 FDG PET/CT in Patients of T Lymphoblastic Leukemia/Lymphoma Treated with ALL-Type Regimens
Authors: Wang Liang , Xia Zhongjun , Chen Xiaoqin , Long Wen
Published: November 2019

Background:

T lymphoblastic leukemia/lymphoma (T-LBL) is highly aggressive. Although intensive chemotherapies such as acute lymphoblastic leukemia (ALL)-type regimens are commonly used, about half adult patients eventually relapse and die of T-LBL. Overwhelming evidences have confirmed the role of interim F-18 FDG PET/CT in Hodgkin lymphoma and many ongoing studies have implemented risk-adapted strategy determined by interim FDG-PET/CT. However, the role of interim FDG-PET/CT in T-LBL remains unclear.

Materials and methods:

47 adult patients of T-LBL treated with ALL-like regimens were retrospectively reviewed. They were treated with modified Berlin-Frankfurt-Münster (BFM)-90 regimen (n=27), Medical Research Council (MRC) UKALLXII/Eastern Cooperative Oncology Group (ECOG) 2993 regimen (n=5), hyperCVAD/MA regimen (n=12), or other unspecified regimens (n=3). Interim (defined as the period between induction therapy and re-induction therapy) FDG-PET/CT was done in all 47 patients and evaluated according to the International Harmonization Project (IHP) criteria.

Results:

The male to female ratio was 3:1, and 29.8% of patients were older than 30 years old. About 90% of patients had mediastinal mass at presentation and 53.2% had bone marrow infiltration. 85.1% of patients had advanced disease. After induction therapy, interim FDG-PET/CT was positive in 19 patients (40.4%), most of whom had residual disease in the mediastinal lesion. Subsequent treatments were not changed according to this interim PET/CT results. After a median follow up time of 28 months, the 2-year and 3-year progression free survival (PFS) rate were 39% and 30%, respectively, and the 2-year and 3-year overall survival (OS) rate were 54% and 45%, respectively. Using Kaplan-Meier survival analysis and log-rank test, it was found that interim FDG-PET/CT positivity correlated with significantly inferior PFS and OS (p=0.002 and 0.010, respectively). Furthermore, patients with higher age (>30) had inferior PFS and OS than younger patients (p=0.037 and 0.036, respectively). However, there were no significant relationship between PFS, OS and bone marrow infiltration, LDH level, and stages (p>0.05).

Conclusions:

Interim FDG-PET/CT may predict PFS and OS in adult patients of T-LBL treated with ALL-type regimens, which needs to be validated in prospective clinical trials. The optimal criteria for interim FDG-PET/CT evaluation and risk-adapted treatment strategy determined by interim FDG-PET/CT should be investigated in future clinical practice.

Disclosures

No relevant conflicts of interest to declare.
Detection of T-Cell Clonality in Bone Marrow in Peripheral T-Cell Lymphoma, Not Otherwise Specified
Authors: Chernova Natalia G. , Sidorova Yulia V. , Ryzhikova Natalia V. , Smirnova Svetlana Yu. , Julhakyan Hunan L. , Sudarikov Andrey B. , Vinogradova Yulia E. , Kovrigina Alla M. , Zvonkov Eugene E. , Parovichnikova Elena N. , Savchenko Valeri G.
Published: November 2019

Introduction: Peripheral T-cell lymphoma, unspecified (PTCL, NOS) is a tumor of mature T-lymphocytes characterized by aggressive course and low response rate to CHOP-like therapy. Bone marrow involvement in PTCL, NOS is detected by histological investigation in about 20-40% of patients and it is considered to be a poor prognostic factor. Importance of T-cell clonality examination in bone marrow as a prognostic and staging factor has been insufficiently studied.

Aim: In order to evaluate the significance T-cell receptor gene γ- and β-chains (TCRG and TCRB) rearrangements for identification of bone marrow involvement; to estimate its value for staging and prognosis; to evaluate the significance of T-cell clonality persistence in bone marrow after initial therapy in patients with PTCL, NOS.

Patients and methods: Bone marrow samples of 27 patients with primary PTCL, NOS (median age 62 years, range 32-77) have been obtained from 2006 till 2014 and were analyzed retrospectively. Morphological examination was performed for all bone marrow samples. Detection of T-cell clonality by revealing rearrangements of TCRB and TCRG were performed by PCR in bone marrow samples of patients before and after initial chemotherapy.

Results: Bone marrow involvement was revealed by histological investigation in 22 (81%) of patients. T-cell clonality was detected by rearrangements of TCRG or TCRB in 26 (96%) patients, including 4 of 5 patients without bone marrow involvement by histology. Rearrangements of TCRG and TCRB were detected in 24 (89%) and 26 (96%) of patients, respectively. Rearrangements of TCRB were observed without rearrangements of TCRG in 2 cases. Interestingly, we detected more than 2 clonal rearrangements present simultaneously in 33% of cases. For monitoring minimal residual disease the examination of T-cell clonality in bone marrow samples was performed in 15 of 27 patients after initial therapy; all these 15 patients achieved complete remission. Persistence of T-cell clonality was observed in 14 of 15 patients after treatment, disease progression was noted in 1-3 months in all these cases.

Conclusion: The examination of T-cell clonality in the bone marrow seems to be important and necessary for staging, monitoring minimal residual disease. Detection of T-cell clonality in bone marrow allows revealing its involvement even without morphological features in primary patients with PTCL, NOS. Persistence of clonal rearrangements in bone marrow after initial therapy shows continuing lesion and necessity for timely therapy escalation for patients with PTCL, NOS.

Disclosures

No relevant conflicts of interest to declare.
High Rates of HTLV-1 Seropositivity in Patients with EBV-Positive Diffuse Large B-Cell Lymphoma
Authors: Beltran Brady E , Morales Domingo , Quinones Pilar , Malaga Jose M , Castillo Jorge J
Published: November 2019

Background: Epstein-Barr virus (EBV)-positive diffuse large B cell lymphoma (DLBCL) of the elderly is a novel entity included in the most recent edition of WHO Classification of hematologic neoplasms. It usually affects patients older than 60 years of age and carries a poor survival. Human T-lymphotrophic virus type 1 (HTLV-1) is a retrovirus that induces immunosuppression, has been associated with the development of adult T-cell leukemia/lymphoma, and has a high prevalence in Peruvian population.

Aim: The goal of this retrospective study was to evaluate the HTLV-1 serological status in EBV-positive DLBCL Peruvian patients.

Methods: We identified untreated patients with a diagnosis of DLBCL between January 2002 and June 2015 from the medical records of our institution. IRB approval was obtained prior to data collection. HTLV-1 positivity was defined based on Western Blot analysis. EBV-positivity was defined by identifying at least 10% EBV-encoded RNA (EBER) expression by chromogenic in situ hybridization in the pathological sample. Clinical data were reviewed retrospectively and patient's biopsies were analyzed for the immunohistochemical expression of CD20, PAX5, BCL6, CD10, CD30 and MUM-1/IRF4 for re-classification purposes.

Results: A total of 210 patients with DLBCL were identified. Of these, 11 patients (5%) were HTLV-1 seropositive and 199 (95%) were HTLV-1 seronegative. Thirty-two patients (15%) were EBV-positive by expression of EBER and 178 (85%) were EBV-negative. HTLV-1 positivity was found in 4/32 (12.5%) of EBV-positive DLBCL patients, and in 7/178 (4%). The odds ratio for HTLV-1 positivity in EBV-positive DLBCL was 3.5 (95% CI 0.7-14.7; p=0.045) when compared with the EBV-negative group. The median age of the HTLV-1-positive DLBCL patients was 65 years (range 45-85 years). There were 5 (45%) men and 6 women (55%). ECOG was >1 in 5 patients (45%), LDH was elevated in 8 (73%), extranodal disease in 7 (64%), and advanced clinical stage (III and IV) in 7 (64%). High IPI score (3-5 factors present) was seen in 8 patients (73%). Interestingly, all the patients who were EBV-positive and HTLV-1-positive presented with early stage (I and II), normal LDH levels and low IPI scores (0-2 factors).

Conclusions: There is a high prevalence of HTLV-1 seropositive status in patients with EBV-positive DLBCL. Patients with double EBV and HTLV-1 positivity appear to present at earlier stages and with lower risk disease. Larger studies are needed to confirm these findings.

Disclosures

No relevant conflicts of interest to declare.
Diffuse Large B-Cell Lymphoma Cell of Origin Determination from Formalin-Fixed Paraffin-Embedded Tissues
Authors: Xu-Monette Zijun Yidan , Thompson Debrah , LaFleur Bonnie , Roche Patrick , Reinholz Monica , Wineman John , Young Ken H. , Botros Ihab
Published: November 2019

Diffuse large B-cell lymphoma (DLBCL) consists of two distinct subtypes, Germinal Center B-cell (GCB) and Activated B-Cell (ABC), each with distinct prognostic and biological profiles. With the advent of new targeted therapeutic approaches for DLBCL treatment, molecular sub-typing of the patient's tumor has been proposed as an important step in therapeutic selection and recommended by 2016 WHO classification guideline.

Gene expression-profiling (GEP) based subtyping is considered the gold-standard method for DLBCL molecular sub-typing, but this method was established using fresh frozen / non-fixed tissues on microarray platforms. Since fresh tissue is not routinely collected during patient diagnosis, it severely limits the ability to utilize this assay in routine clinical practice.

Multiple immunohistochemical (IHC) approaches have been developed to approximate the GEP methods, but these techniques generally suffer from lower than desired agreement rates with GEP classifications, and require staining of 4 to 8 sections of limited biopsy material. Interpretation of the slides can also be variable, leading to low inter-observer reproducibility.

We have developed a 12 gene GEP-based DLBCL cell-of-origin (COO) assay using the HTG EdgeSeq System specifically designed to use a minimal amount of FFPE tissue. To build this system, we profiled a total of 107 samples previously subtyped using the HG-U133 Plus 2.0 Affymetrix microarrray and algorithm (Visco C et al Leukemia 2013); this algorithm was then validated in an additional 58 samples. The methodology we have developed produces 92% concordance with the microarray-based approach.

Briefly, 107 DLBCL cases, of which 58 were previously sub-typed as ABC and 49 cases as GCB, were used as the training cohort. After classifier training and cross-validation, a separate cohort of 58 cases were used to verify the performance of the assay/classification system. Approximately 5 mm2 of 5 µm thick FFPE tissue was used to generate the data set for each of the cases.

In addition to the DLBCL COO classification, the assay also contains additional genes including potential drug targets, T-cell, B-cell, and macrophage biomarkers, and housekeeping/normalization genes. These markers could be used to further understand the nature of the tumor and potentially help identify the characteristics of atypical tumors and immune infiltrates in the microenvironment.

Disclosures

Thompson: HTG Molecular Diagnostics, Inc: Equity Ownership. LaFleur:HTG Molecular Diagnostics, Inc: Equity Ownership. Roche:HTG Molecular Diagnostics, Inc: Equity Ownership. Reinholz:HTG Molecular Diagnostics, Inc: Equity Ownership. Wineman:HTG Molecular Diagnostics, Inc: Equity Ownership. Botros:HTG Molecular Inc: Equity Ownership.
Hodgkin's Lymphoma Outcome: A Retrospective Study from Three Tertiary Centers in Saudi Arabia
Authors: Shafi Ruaa , Al-Mansour Mubarak M , Kanfar Solaf Sami , Al Hashmi Hani Hassan , Alsaeed Ahmad S. , Al-Foheidi Meteb , Ibrahim Ezzeldin
Published: November 2019

Background: Hodgkin lymphoma (HL) demonstrates considerable clinicopathologic variations in different parts of the world. Prompted by the limited availability of data particularly the long-term outcome of HL in developing countries, we carried out this retrospective data analyses.

Methods: A retrospective review of eligible adult HL patients managed at three tertiary centers in Saudi Arabia between January 1997 to December 2012.

Results: The review included 340 patients with median age of 26 years (range15-82). 53% of patients were male, 74% had an advanced stage, 19% had bulky disease, and 70% had low to intermediate risk according to Hasenclever index. Nodular sclerosis was the most common histological subtype (59%). ABVD was given to 92% and radiotherapy to 43%. Response to initial therapy was complete, partial, and disease progression was reported in 91%, 5%, and 2 % of patients, respectively. At a median follow-up of 50 months, the actuarial freedom from treatment-failure at 5-year was 74%, and with an overall survival of 91%. Multivariate analysis showed that advanced disease stage and high-risk international prognostic index independently projected an adverse outcome.

Conclusion: This study confirms that our patients population demonstrated a comparable outcome to that reported from developed countries.

Disclosures

No relevant conflicts of interest to declare.