bluuebunny/crossref_metadata_2025_split huggingface dataset:
Clinical Impact of Immunophenotype-CD15/CD30 on Outcome of Hodgkin Lympoma
Authors: Zafar Huma , Riaz Shazia , Badar Farhana 
Published: November 2019

Abstract
Introduction
In classical Hodgkin Disease, Reed Sternberg cells reacts with antibodies against a granulocyte associated antigen, CD15/CD30. So expression of immunochemistries CD15 and CD 30 in patients,suspecting for HD, has quite significant diagnostic value.
Aim
To find out relevance of CD15/CD30 expression in relation to clinical outcome in patients of Hodgkin's disease.
Methods and Material
A total of 301 patients of Hodgkin' disease from stage I to stage IV treated in Shaukat Khanum hospital during last 8 years, were reviewed. Data was analyzed concerning age, gender, histopathology, staging, immunochemistry and outcome.
Results:
Among these 301 patients, male to female ratio was 4.3:1.The mean age was 10.01years Maximum patients presented in stage III i.e 40.2 %. Total 55 (18.2%) patients were reported positive for CD 15 and 261 (86.7 %) for CD 30. Amongst CD 15 positive patients, 1.8% received palliation, 16.3% showed Primary progressive Hodgkin's lymphoma ( PPHL ),16.3% showed relapse. Amongst CD 15 negative patients, no patients received palliation, 9.47% showed PPHL, 15.7% showed relapse.
Amongst CD 30 positive patients, 0.38% pts received palliation, 7.66% showed PPHL, 12.2% showed relapse.
Conclusion:
There is diagnostic significance of CD15/CD 30 in Hodgkin's Disease but the prognostic significance of these immunochemistries could not be proven.

Disclosures
No relevant conflicts of interest to declare.
Suppression of EZH2 Accelerates MYC-Driven Lymphomagenesis By Inhibition of Apoptosis
Authors: Appelmann Iris , Scuoppo Claudio , Thapar Vishal , Ledezma Daniela , Lujambio Amaia , Lowe Scott W. , Chicas Agustin 
Published: November 2019

Abstract
EZH2 is the catalytic component of the polycomb repressive complex 2 (PRC2), which also contains the non-catalytic subunits suppressor of zeste 12 (SUZ12) and embryonic ectoderm development (EED). This complex methylates histone H3 at lysine 27 (H3K27) which, together with H3K4 methylation, generates a bivalent “code” that primes genes for either expression or silencing. EZH2 is highly expressed in stem cells and many proliferating cells, downregulated in differentiated cells and frequently altered in cancer in ways that point to a context dependent role for this gene. For instance, overexpression of EZH2 has been described in prostate and breast cancer, where this overexpression is associated with invasive growth metastatic potential and poor clinical outcome.
More recently, both gain and loss of function mutations of EZH2 were identified in human cancer. In particular, activating mutations of EZH2 affecting a tyrosine at position 641 located within the SET domain (Y641) were observed in lymphoma. In striking contrast to wild type EZH2 which catalyzes the monomethylation of H3K27 very efficiently and shows less efficient catalytic activity in the subsequent di- and trimethylation reactions, the mutation generates a neomorphic protein with enhanced catalytic activity and efficiency for di- and tri-methylation, hinting toward a “cooperation” of both wild type and mutant EZH2 to increase total H3K27me3 levels and representing a functional equivalent of EZH2 overexpression in human lymphoma.
Gain of function mutations in EZH2 are frequent in Diffuse Large B-cell Lymphoma (DLBCL; 22%) and in Follicular Lymphoma (FL; 7%) and recent data implicate EZH2 as an oncogene in DLBCL and FL lymphomas.
In contrast to DLBCL and FL, EZH2 mutations have so far never been identified in MYC-driven B-cell Non-Hodgkin Lymphoma (B-NHL), and EZH2 expression is suppressed in Burkitt’s Lymphoma (BL), a lymphoma for which a MYC translocation is pathognomonic, suggesting that in this context EZH2 could have a role different from its role in DLBCL and FL. We probed the role of EZH2 in MYC-driven lymphomagenesis by mimicking the loss of function mutations by RNAi mediated suppression of EZH2 and the gain of function mutations by over-expression of the Y641 mutant.
Our results show that suppression of EZH2, but not overexpression of the EZH2 Y641 mutant, accelerates MYC-driven lymphomagenesis by attenuating apoptosis. Our model recapitulates the transcriptional signature of a subset of B-NHLs driven by MYC overactivation and EZH2 suppression. Taken together, our data imply EZH2 as a tumor suppressor in the context of MYC activation and thus raise a warning for the use of EZH2-targeted therapies in some B-NHLs subtypes.

Disclosures
No relevant conflicts of interest to declare.
Predictive Value of Interim 18f-FDG PET-CT Scans on Diffuse Large B-Cell Lymphoma Treated with R-CHOP: A Prospective Study of Chinese Southwest Oncology Group (CSWOG)
Authors: Huang He , Lin Jia Tian , Guo Chengcheng , Hong Huangming , Prem Raj Shrestha , Li Xueying , Zhang Mengping , Li Shanshan , Fang Xiaojie , Wang Zhao , Xia Zhongjun , Lin Tongyu 
Published: November 2019

Abstract
Purpose
18F-FDG PET-CT has been widely used for pre-treatment staging and post-treatment response assessment in diffuse large B cell lymphoma (DLBCL), but the predictive value of interim PET-CT remained controversial and most of studies were retrospective.
Patients and Methods
Newly-diagnosed DLBCL patients treated with R-CHOP regimen were included in our prospective study to evaluate the predictive value of interim PET-CT. All patients were evaluated with PET-CT scans before treatment and after every 2 cycles of R-CHOP. PET-CT positivity or negativity was related to survival using Kaplan-Meier analysis.
Results
From Feb 2008 to Jan 2013, 149 patients were included. After 2 cycles of R-CHOP, the PET-CT evaluation showed CR in 82 patients. Among the remaining 67 non-CR patients, 31 achieved CR after 4 cycles. At the end of treatment, PET-CT evaluation showed CR in 121, PR in 21, SD in 3 and PD in 4 patients. With a median follow-up of 20.6 months (range 1.5-60.5 months), patients with negative PET-2 (PET-CT scan after 2 cycles) had a superior 2-year PFS than those with positive PET-2 (86.6% vs. 67.0%, p=0.019) and a tendency of superior 2-year OS without statistical differences (91.9% vs. 85.2%, p=0.330). The 2-year PFS and OS for negative PET-4 (PET-CT scan after 4 cycles) compared with positive PET-4 group were 84.8% vs. 51.9% (p=0.001) and 93.1% vs. 73.0% (p=0.027) respectively. PET-CT scans were interpreted using the International Harmonization Project (IHP) criteria above. The second analysis applying the Five-Point Scale (5PS) criteria showed that 2-year PFS for score 1-2 (uptake＜mediastinum), score 3 (mediastinum＜uptake≤liver) and score 4-5 (uptake＞liver) group were 86.6%, 75.8% and 71.0% according to PET-2, and 84.9%, 50.0% and 52.7% according to PET-4. PFS of score 3 group was not significant different from the other two groups in PET-2 (both P＞0.05), but significantly inferior to score 1-2 group (p=0.046) and similar to score 4-5 group (p=0.767) in PET-4. In the multivariate analysis, only PET-4 (95%CI, 1.07-5.44) and IPI score (95%CI, 1.53-10.06) remained independent predictive factors for PFS.
Conclusion
PET-CT after 4 cycles of R-CHOP in patients with DLBCL is highly predictive of PFS and should be considered in clinical practice.
(Clinical Trail Registration Number: CTR-TRC-11001687)

Disclosures
No relevant conflicts of interest to declare.
Identification of Cell-Type-Specific Mutations in Angioimmunoblastic T-Cell Lymphoma
Authors: Nguyen Tran Bich , Sakata-Yanagimoto Mamiko , Asabe Yukitsugu , Yoshida Kenichi , Muto Hideharu , Nakamoto-Matsubara Rie , Izutsu Koji , Nakamura Naoya , Takeuchi Kengo , Ogawa Seishi , Chiba Shigeru 
Published: November 2019

Abstract
[Backgrounds] Angioimmunoblastic T-cell lymphoma (AITL) is a distinct subtype of peripheral T-cell lymphoma (PTCL), characterized by generalized lymphadenopathy and autoimmune-like manifestations. Regarding genetic lesions of AITL, frequent mutations in TET2, IDH2, DNMT3A and RHOA have been identified. In some PTCL cases, TET2 and DNMT3A mutations were identified in cell populations beyond the CD4+ T-lymphocytes, in which the tumor cells are contained, suggesting that TET2 and DNMT3A mutations occurred earlier than the commitment to CD4+ T lymphocytes.
[Objective] We performed this study to identify the cell-type-specific mutations and further explore mutational profiles in AITL and AITL-related cancer.
[Methods] The dataset of targeted sequencing was analyzed for 76 genes in 79 PTCL samples. Mutational origin was analyzed by cell sorting and laser microdissection.
[Results] Targeted sequencing identified 168 mutations in 33 genes. Recurrent mutations, in addition to the already known frequent mutations in RHOA/TET2/IDH2/DNMT3A, were found in ODZ1 [4/79 (5%)], Notch1, NAV2, and MTERFD3 [3/79 (4%) for each], MLL2, TET3, FAT2, and LAMA2 [2/79 (3%) for each]. TET2/DMNT3A mutations showed statistically higher allelic burden than the newly identified mutations, suggesting precedence of TET2/DNMT3A mutations.
Cell sorting and laser microdissection, followed by amplicon sequencing, revealed that TET2/DNMT3A mutations were identified in both tumor cell-enriched and –depleted populations while RHOA and IDH2 mutations were confined to tumor cell-enriched populations. Most of the newly identified mutations were similarly classified into the above-mentioned two types. It is noteworthy that we found some mutations only in T-cell lymphoma cell-depleted CD20-positive population but not in the tumor-cell-enriched PD-1-positive population.
[Conclusion and discussion] Differentiation stages that mutational events arise are likely to be multiple in AITL and AITL-related lymphoma. Moreover, in AITL, Epstein-Bar virus-infected B cells often grow in an oligoclonal manner, sometimes resulting in monoclonal proliferation with fully malignant features. Detection of B-cell specific mutations might suggest premalignant status of B cells in these cases.

Disclosures
No relevant conflicts of interest to declare.
Clinical Prognostic Models in Diffuse Large B-Cell Lymphoma Patients Are Still Essential in the Rituximab Era
Authors: Lage Luis Alberto de Padua Covas , Costa Renata Oliveira , Hallack Neto Abrahão Elias , Siqueira Sheila , Santucci Rodrigo , de Paula Henrique Moura , Pereira Juliana 
Published: November 2019

Abstract
Introduction: To evaluate a new enhanced IPI proposed by the National Comprehensive Cancer Network (NCCN-IPI) in DLBCL patients, we compared the international prognostic index (IPI), R-IPI and NCCN-IPI in DLBCL patients treated with rituximab, cyclophosphamide, hidroxydaunorubicin, vincristine and prednisone (R-CHOP).
Methods: From June 2008 to November 2011 we retrospectively evaluated 146 DLBCL patients treated with R-CHOP-21 referred for cancer treatment in a single university institution in Brazil. Patient's clinical data were assessed to calculate the IPI, R-IPI and NCCN-IPI.
Results: Patient's median age was 58.9 years (range 16 – 86); 85 (57.8%) were female. According to IPI, risk categories were low (n=41, 28.1%), low-intermediate (n=43, 29.5%), high-intermediate (n=37, 25.3%) and high (n=25, 17.1%). Using R-IPI, risk categories were very good (n=19, 13%), good (n=65, 44.5%) and poor (n=62, 42.5%). According to NCCN-IPI, risk categories were low (n=12, 8.2%), low-intermediate (n=52, 35.6%), high-intermediate (n=62, 42.5%) and high (n=20, 13.7%). At 30 months (median follow up 17.7 months - range 0.6-58.2 months) the overall survival (OS) was 75.5%. The progression-free survival (PFS) at a median follow-up of 16.3 months (range 0.6-52.4) was 68.3% for all patients. Using IPI, the OS at 30 months did not differ between low and low-intermediate risk patients (96.8% vs. 82.2%; p=0.136); however, it was higher than the OS of high-intermediate risk (n=37; 96.8% vs 74.1% p=0.11) and high-risk (n=25; 96.8% vs 41% p < 0.001) patients (Figure 1). The NCCN-IPI demonstrated significant differences in OS (p < 0,001) and PFS (p<0.001) among low-intermediate, high-intermediate, and high risk groups, with the high-risk group exhibiting worse OS (32.1% in 30 months) (Figure 2). According to IPI, the OS in high-risk patients was 41%.
Figure 1: OS and PFS according to International Prognostic Index (IPI) Figure 1:. OS and PFS according to International Prognostic Index (IPI)
Figure 2: OS and PFS according to NCCN-IPI Figure 2:. OS and PFS according to NCCN-IPI
Figure 3 Figure 3.
Conclusion: In our study the NCCN-IPI, but not the IPI or R-IPI was able to discriminate a high-risk group of DLBCL patients treated with R-CHOP with worse OS.

Disclosures
No relevant conflicts of interest to declare.
Brentuximab Vedotin Followed By ABVD in Patients with Previously Untreated Hodgkin Lymphoma. a Pilot Phase II Study
Authors: Zinzani Pier Luigi , Luminari Stefano , Merli Francesco , Pesce Emanuela Anna , Chauvie Stephane , Pellegrini Cinzia , Marcheselli Luigi , Capodanno Isabella , Ilariucci Fiorella , Salati Massimiliano , Argnani Lisa , Federico Massimo 
Published: November 2019

Abstract
active and well tolerated single agent in the treatment of heavily pretreated Hodgkin lymphoma patients. In this pilot phase II study patients with previously untreated HL underwent sequential regimen consisting in 2 cycles of BV before doxorubicin, bleomycin, vinblastine and dacarbazine (ABVD) +/- radiotherapy (RT). The primary endpoint of the study was the response to BV assessed by positron emission tomography (PET) after the 2 cycles (PET2), defined as reduction of Deauville score or, in case of no change in Deauville score, as any reduction in standard uptake value (SUV) intensity compared to basal SUV.
Between April and October 2013, 12 patients with a median age at diagnosis of 36 years (range, 19-70), 11 stage I-IIA and 1 stage IIIA, were enrolled. BV was administered as scheduled and at the full dose of 1.8 mg/kg in all patients. After the 2 cycles of BV, the overall response rate was 91%, comprising of ten (83%) complete responses and one (8%) partial metabolic response. The non responding patient had stage III and showed a new lesion at PET2. After ABVD +/- RT, the overall response rate was 100% with 11 complete responses and 1 partial response (converting from progression disease). At a median follow up of 8 months, all 11 patients are still in complete response, while the remaining one relapsed.
During BV therapy, the only grade 3 adverse events were transient and asymptomatic increase in liver transaminases (n=3, 25%) and gamma glutamyl transpeptidase (n=2, 17%). During ABVD +/- RT grade III-IV neutropenia occurred in 9 (75%) patients. All toxicities were transient and resolved with growth factor support.
Two cycles of BV as first line-treatment in limited stage HL induced an outstanding complete response rate with limited toxicity and reducing the need of chemotherapy. Waiting for a longer follow up to assess the duration of response, our data (sequential administration of immunotherapy and chemotherapy) should be considered the starting point for further studies in first line therapy for limited stage HL patients.

Disclosures
No relevant conflicts of interest to declare.
Germ-Line and Somatic Mutations in Familial Myeloproliferative Neoplasms (MPNs). a Pilot Study
Authors: Nand Sucha , Diaz Manuel O. , Marinier David E. , Walshauser Mark A. , Rosenfeld Amy B , Michaelis Laura C , Stiff Patrick J , Gai Xiaowu 
Published: November 2019

Abstract
Background: Familial MPNs are uncommon disorders that, like sporadic cases, are characterized by clonal hematopoiesis and presence of somatic mutations, e.g. JAK2, CALR, MPL and occasionally TET2. There is little information, however, about germ-line mutations in these families that may explain the low penetrance hereditary predisposition.
Methods: We studied five families with MPNs, each with at least 2 affected members. After obtaining an informed consent, clinical data was obtained from the patients’ electronic medical records. Blood and buccal samples were collected from patients and unaffected relatives. Exome sequencing was performed on the blood DNA samples using Agilent SureSelect Human All Exon V5+UTRs exome capture kit followed by massively parallel sequencing with Illumina HiSeq 2000. Sanger sequencing was then done on both the blood and buccal swab DNA samples to validate selected gene variants and to differentiate the nature of those variants (germ line or somatic).
Results: The 5 families participating in this study had the following diagnoses: 1. Mother: polycythemia vera (PV); son: essential thrombocythemia (ET), 2. Mother: primary myelofibrosis (MF); daughter: unclassifiable MPN (UMPN), 3. Father: PV; son: PV, 4. Sister: MF; sister: MF, 5. Two aunts: MF; niece; UMPN. Six patients were positive for JAK2, V617F mutation. Blood and buccal samples were collected from 5 patients and 4 relatives. In all 5 families, the pro-band was younger at the time of diagnosis than his/her affected relatives. The clinical course of the MPNs appeared to be similar to the sporadic form.
Exome sequencing revealed TET2 mutations in 2 probands. In addition, novel non-synonymous mutations in several candidate genes, KMT2D, KMT2C, NBEAL1, NBEAL2, AHNAK2, RNF213, were identified in the blood samples from the patients but not their unaffected relatives. These include two novel KMT2D mutations in two unrelated families. These 2 mutations were also found in the matching buccal swab samples, indicating that they are germ line mutations.
Discussion: KMT2D and KMT2C mutations have been previously identified as somatic mutations in lymphoid malignancies, including non-Hodgkin’s lymphomas (Morin 2011), and as germ line compound heterozygote mutations in infant MLL and ALL (Valentine 2014). About 32% of diffuse large cell lymphoma and 89% of follicular lymphoma have somatic mutations of KMT2D. NBEAL2 germ line mutations are associated with familial gray platelet syndrome, where some patients have myelofibrosis (Gunay-Aygun 2011). To our knowledge, this is the first report describing germ line mutations in familial MPNs. The possible role of these mutations in predisposition to MPN will be discussed. Studies on additional families with MPNs are planned.

Disclosures
No relevant conflicts of interest to declare.
Protooncogene MYB Activates the Transcription of ROR1 in B Cell Malignancies
Authors: Watanabe Ken , Iida Megumi , Suwa Shihoko , Kato Miyu , Miura Osamu , Fukuda Tetsuya 
Published: November 2019

Abstract
The receptor tyrosine kinase-like orphan receptor 1 (ROR1) is a highly conserved gene that is associated with cell survival. ROR1 is uniquely expressed in some B cell malignancies and epithelial cancer cells although it is barely detected in normal adult tissues. The precise mechanisms of this ectopic expression have remained unknown. We checked ROR1 expression by FACS analysis and by quantitative RT-PCR. ROR1 was highly expressed in the samples from CLL,MCL,SMZL,and BL. Expression levels of ROR1 mRNA and cell surface ROR1 protein showed clear positive correlation (R2=0.55) suggesting that ROR1 expression is regulated mainly transcriptionally. Then, we cloned the 2.5-kb fragment upstream of ROR1 transcriptional start site and performed promoter assay using ROR1 positive and negative cell lines. The promoter activity was observed not only in ROR1 positive lines but also in negative lines, which suggests that this region may not be the only regulatory element. Thus, we hypothesized that conserved regions in intron 1, which is about 230 kb long, may play important roles in regulation of transcription. BLAST search identified 19 regions conserved between human and rat intron 1 of ROR1. Two regions, designated as CRS3 and CRS7, further showed homology with chicken ROR1 gene. Among them, CRS3 region had enhancer activity selectively in ROR1 positive B cell lines. A consensus nucleotide sequence for the binding of protooncogene MYB was found within CRS3 region. Actually, the protein binding to CR3S sequence was verified by EMSA assay using nuclear extract from MYB positive cells. Immunoblot analysis showed that ROR1 positive B cell lines had high expression of MYB. In addition, the expression levels of MYB protein and ROR1 mRNA were positively correlated in 7 CLL samples analyzed (R2=0.49). We introduced shRNA of MYB into ROR1 positive EW36 cell line. MYB shRNA could decrease CRS3 luciferase reporter activity. As a result, MYB shRNA decreased ROR1 expression as well as MYB expression. Collectively, MYB induced ROR1 expression and might contribute to tumorgenesis of B cell malignancies.

Disclosures
No relevant conflicts of interest to declare.
Hypermethylation of TAp73 Suppresses ABL1-Involved DNA Damage Response in Multiple Myeloma
Authors: Hao Mu , Qin Yu , Zang Meirong , Xu Yan , An Gang , Li Changhong , Yang Ye , Gu Zhimin , Zhan Fenghuang , Qiu Lugui 
Published: November 2019

Abstract
Background: More recently, multiple myeloma (MM) cells evade apoptosis despite pervasive DNA damage was demonstrated. However, the relevance of ongoing DNA damage and the mechanisms by which apoptosis is suppressed remain to be fully elucidated. p53 deletion and mutations do not appear to be a pivotal event in the evolution from pre-malignancy toward malignancy in MM. The protooncogene ABL1 was an alternative pathway to p53 down stream of ATM/ATR, which is commonly translocated in Chronic Myelogenous Leukemia (CML). ABL1 forms a complex with the tumor suppressor gene TP73, which belongs to the p53 family. P73 is expressed as multiple isoforms due to the usage of two different promoters, P1 promoter of TAp73 inducing apoptosis and P2 promoter of Δ Np73 promoting survival. In this study, we explored the role of ABL1/p73 axis in MM cells evading ongoing DNA damage induced apoptosis.
Materials and methods: Real-time-PCR was used to detect the ABL1 and miR-203a expression in MM primary samples and MM cell lines. Immunofluency staining was performed to detect the ɣ-H2A.X level in MM cells. Flow cytometry was performed to detect the apoptosis in MM cells. Bisulfite Pyrosequencing and Methylation Specific-PCR were used to detect the p73 promoter methylation.
Results: Our results revealed that ABL1 level was up-regulated both in primary MM samples and MM cell lines (-1.25±0.28 vs. 0.06±0.24, p=0.02). MiR-203 which suppresses ABL1 expression was down-regulated (0.01±0.01 vs. 0.97±0.08, p=0.01). MM cell lines and primary cells showed high ɣ-H2A.X staining. Immunofluency staining showed that ABL1 relocalized in the nucleus of MM cells after treated with doxorubicin. The apoptosis of MM cells was significantly up-regulated (7.8±2.1)% vs. (25.4±4.5)%, p<0.05. Doxorubicin treatment combined with ABL1 inhibitor (STI571) suppressed the apoptosis significantly, (25.4±4.5)% vs. (12.2±3.4)%, p=0.03. Bisulfite Pyrosequencing and MS-PCR of 42 newly diagnosed MM patient sample revealed that the P1 promoter of p73 was hypermethylated compared with normal plasma cells (86% ±7% vs. 58%±4%, p=0.032). RT-qPCR and western blotting showed that Δ Np73 levels were significantly higher than TAp73 (148.6±19.3 vs. 6.8±2.4, p=0.021) in plasma cells of those patients and MM cell lines.
Conclusion: Hypermethylation of p73 promoter suppresses TAp73 expression. The deregulated of ABL1-p73 pathway in MM cells resulted in marked reduction of apoptosis induced by ongoing DNA damage.

Disclosures
No relevant conflicts of interest to declare.