bluuebunny/crossref_metadata_2025_split huggingface dataset:
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

Abstract
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

Abstract
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

Abstract
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

Abstract
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

Abstract
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

Abstract
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

Abstract
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.
Pregnancy and Primary Mediastinal B-Cell Lymphoma
Authors: Mangasarova Jana Konstantinovna , Kravchenko Sergey Kirillovich , Magomedova Aminat Umaraskhabovna , Baryakh Elena , Vorobiev Vladimir Ivanovich , Savchenko Valeri G. 
Published: November 2019

Abstract
Introduction. Favorable therapeutic regimen for pregnant women with primary mediastinal B-cell
Lymphoma (PMBCL) is determined by tumor mass, somatic status, period of pregnancy and potential risk of teratogenic effects of medications which penetrate through placenta.
Aim. To estimate chemotherapy efficacy of PMBCL in case of pregnancy and its toxicity for fetus.
Patients and Methods. Since 2004 to 2015 years in National Research Center for Hematology were treated 106 patients with primary mediastinal B-cell lymphoma. In 8 (7,5%) from them disease debuted during 2-3 trimester of pregnancy, one women had two fetuses simultaneously. Before delivery women underwent induction chemotherapy with VACOPB- regimen in 5/8 cases and with R+DAEPOCH in 3/8 cases.
Results. After induction treatment had been performed 6/8 women achieved a partial remission, 1/8 had progression, 1/8 - complete remission. High dose chemotherapy was performed in 7/8 women after 1 month following delivery. It was born 9 children (4 boys and 5 girls). Median body weight was 2000 g. (1300 - 3600 g.). Median growth was 47 cm. (40 - 53 cm). In two cases when rituximab had been administered to women before delivery, children were diagnosed with intrauterine pneumonia. One child, born from woman after VACOPB chemotherapy had thrombosis of superior vena cava. All women continue to be in complete remission of PMBCL, children are practically healthy without malformations. The median follow-up of patients was 40 months (15 - 78 months).
Conclusions. Pregnancy is not a contradiction for induction chemotherapy of PMBCL in 2-3 trimesters. Rituximab shouldn't be administered before delivery because of a high risk of infectious in fetus.

Disclosures
No relevant conflicts of interest to declare.
Retrospective Study of the Efficacy and Safety of Treatment with Subcutaneous Injection of Bortezomib in De Novo Patients with MM
Authors: Fu Chengcheng , Ma Ling , Yan Shuang , Liu Hui , Xu Yun , Depei Wu 
Published: November 2019

Abstract
[Objective]
The induction therapy of multiple myeloma (MM) has dramatically changed due to the availability of new drugs such as bortezomib.To explore the efficacy and safety of subcutaneous injection of bortezomib in the treatment of de novo MM patients, a total of 56 patients treated with PAD form June 2008 to January 2015 were retrospective analyzed.
[Methods]
29 patients in PAD group received intravenous injection of bortezomib 1.3mg/m2, d1,4,8,11, Adr iv 40mg/m2, d1, Dex iv 20mg, d1,2,4,5,8,9,11,12; another 28 in improved PAD group received subcutaneous injection of bortezomib1.3mg/m2, d1,4,8,11, Adr iv 40mg/m2, d1, Dex iv 20mg, d1,2,4,5,8,9,11,12. The efficacy and safety of two groups were analyzed.
[Results]
The response rate >= VGPR after 4 cycles of PAD or improved PAD were 51.7% and 51.9% respectively.With a median follow up of 24 months,the median TTP , PFS and OS of the PAD group was 47 months, 42 months, 63 months respectively, but the time of the improved PAD group wasn't shown up yet.In the improved PAD group,2y- TTP ,PFS and OS was 94.4%, 94.4%, 100%. With a follow-up to 60 months,the OS of standard-risk group[without FISH 1q21+, or t(4;14), or p53-]was 61.9%,but the survival was only 31.9% in FISH high-risk group. OS, PFS of the patients with 1q21 amplification was 18.7-56.4 months and 9.7-53.5 months, showed statistical difference (P=0.029), (P=0.037), compared to FISH negative patients,which was 51.5-71.3 months and 39.4-68.4 months. The study found that the hemoglobin < 100 g/L or 1q21+ were the poor prognosis factors for patients' OS, while subcutaneous injection of bortezomib could improve OS(P=0.042), mainly because compared to PAD group,Grade 3 or worse adverse events were significantly reduced in improved PAD group. the most common were peripheral neuropathy (P=0.049), infection(0.004). When α=0.1, leukopenia(P=0.086), diarrhea(P=0.093) also showed statistically significant difference.
[Conclusions]
The improved PAD regimen by changing bortezomib from intravenous administration to subcutaneous injection significantly reduced adverse events, improved the safety of clinical application of bortezomib without affecting curative effect, and had greatly improved the OS.1q21 amplification was the poor prognostic factors in PAD induction treatment of MM.

Disclosures
No relevant conflicts of interest to declare.