bluuebunny/crossref_metadata_2025 huggingface dataset:
Isolation, Expansion and Characterization of BM-Mesenchimal Cells (MSCs) in Patients Affected By Chronic Myeloproliferative neoplasm (MPN)
Authors: Chiusolo Patrizia , Annunziata Francesca , Rossi Elena , Betti Silvia , Bellesi Silvia , De Stefano Valerio , Leone Giuseppe
Published: October 2019

Introduction MSCs constitute a pivotal cell type capable of shaping both the architecture of the microenvironment and modulating communication between the various cell types through effects on the extracellular matrix (ECM) and by secretion of various growth factors and cytokines. MSCs and hematopoietic stem cells are thought to share the same mesenchymal origin. Some data confirm that MSCs express a functional erythropoietin receptor and JAK2-transduction pathway, but their role in the development and evolution of MPN is still not well known so our aim was the isolation, expansion and characterization of MSCs in patients affected by MPN. Some data indicates that BM-MSCs of patients affected by MPN do not carry the JAK2-V617F mutation. We studied 20 patients affected by MPN with the following characteristics: M/F 12/8, median age 53years, 8 affected by PV, 8 by ET and 4 by PMF. 15 patients were positive for JAK2 V617F mutation, 1 pts for cMPL and 2 were CARL mutations carriers.

Methods: MSC were isolated by bone marrow fraction by gradient separation on Lympholyte cell separation media and expanded in culture with a specific medium (MesenCult) in plastic-adherent cultures up to the second passage. DNA was extracted from MSC using QIAmp DNA Mini kit and the study of recurrent alterations (JAK2, MPL and CARL gene mutations was performed).

Results: MCS were expanded in 14 on 21 patients. Flow citometry analysis confirmed the standard MSC phenotype (CD45 negative, CD73 positive, CD90 positive and CD105 positive). The molecular analysis of JAKV617F, cMPL and CARL mutations resulted negative in all analyzed samples both in patients carriers of mutations and in wild type ones.

Conclusions: We conclude that common mutations markers of MPN neoplasm are absent in the mesenchymal compartment of bone marrows of patients affected by MPN and are restricted to the neoplastic clone.

This research project was supported by a grant from Associazione Italiana per la Ricerca sul Cancro (A.I.R.C.)

Disclosures

No relevant conflicts of interest to declare.
Idiopathic Hepatitis Associated Aplastic Anemia
Authors: Rivera-Trujillo Alicia , Navarro-Escamilla Adolfo , Hernandez-Rodriguez Sonia , Campos-Cabrera Virginia , Campos-Cabrera Gregorio
Published: October 2019

Aplastic anemia (AA) in Mexico has an incidence of 3.5-4.0 x 106 (Hematology 2002;7:229-232). The presentation of idiopathic hepatitis associated aplastic anemia (IHAAA) in México is unknown due to data was not found on PubMed, but represents from 2-5% of all cases of AA in Europe and Far East (Virology 2011;8:87 and Br J Haematol 2010;149:890-895).

A previously healthy and normal family history 15 years old boy presented with epistaxis, pallor, jaundice, choluria and slightly hepato and splenomegaly, which were confirmed with full body CT scan, and no other alterations were detected. Remarkable results from AMA designated disease/organ panels were Hb 11.7 g/dL, adjusted reticulocytes 0.26%, neutrophils 0.931 K/µL, platelets 5.0 K/µL, ALT 1920 U/L, AST 2300 U/L, conjugated bilirubin 17 mg/dL, unconjugated bilirubin 15 md/dL, ferritin 1135 ng/mL. Supportive care was given and study for pancytopenia and hepatitis was started. Negative results from complete autoimmune panel including anti-LKM, anti-smooth muscle and anti-mitochondrial antibodies; panels for all hepatitis viruses, HIV, STORCH, EB, CMV, PVB19 and leptospira were negative. The patient was prepared for biopsies. Liver biopsy showed steatohepatitis, grade 2 fibrosis and lymphocyte infiltrates. Bone marrow biopsy showed 5% cellularity with absence of myeloid and megakaryocytes precursors, only erythroblasts and lymphoplasmocytoid cells; flow cytometry reported no abnormal cells and G-band karyotypes with and without mitomycin were normal. While the results from the biopsies the hepatic function improves but pancytopenia worsened to Hb 6.4 g/dL and neutrophils 0.124 K/µL. IHAAA was diagnosed and the patient continues with supportive care while results from siblings for allogeneic transplant arrive.

IHAAA is well described condition where secondary causes are ruled out. Autoreactive T lymphocytes and cytokines are implied in the pathophysiology as in conventional AA, affecting transiently the liver; and treatment is the same for both entities.

Figure 1 Figure 1.

Figure 2 Figure 2.

Figure 3 Figure 3.

Disclosures

No relevant conflicts of interest to declare.
Aberrant Overexpression and Regulatory Mechanism of Mir-21 in Diffuse Large B Cell Lymphoma
Authors: Fu Rong , Li Chunhong , Shao Zonghong
Published: October 2019

MicroRNAs (miRNAs) are critically responsible for lymphomagenesis. We found that miR-21 expression was significantly higher in diffuse large B cell lymphoma (DLBCL) line and paraffin-embedded tissue of primary lymphoma (n = 26) than that in normal lymph nodes (n = 10). To determine the functions of miR-21 in lymphomagenesis, we examined phosphatase and tensin homolog (PTEN) expression using immunohistochemical SP method in specimens of primary DLBCL. MiR-21 expression is negatively correlated with PTEN expression, but positively correlated with LDH. MiR-21 expression was significantly higher in stage III / IV patients than in stage I/II patients. Moreover, we examined the effects of antisense oligonucleotide (ASO) targeting miR-21 (ASO-21) in the DLCBL line overexpressing this miRNA. Electroporation was used to transfect ASO-21 into the cells, which showed slight endogenous miR-21 expression. Downregulating miR-21 expression resulted in increased incidence of apoptosis and PTEN upregulation. These results provide relevant new insight into the pathogenesis of DLBCL and suggest that targeting miR-21 may represent a useful approach for DLBCL treatment, and that high levels of miR-21 may be associated with increased poor prognosis for DLBCL patients.

Disclosures

No relevant conflicts of interest to declare.
Treatment of Acute Myeloid Leukemia with the NOPHO-AML 2004 Study in Chinese Children: A Single Center Study
Authors: Feng Xiaoqin , Li Chunfu
Published: October 2019

Objectives: The objective of the present study was to investigate the therapeutic efficacy and feasibility of NOPHO-AML 2004 study in the treatment of acute myeloid leukemia (AML; excluding acute promyelocytic leukemia) in Chinese children.

Methods: Thirty-one children with novo AML treated with the NOPHO-AML 2004 study were recruited from Jan. 2010 to Dec. 2013, and the clinical data were retrospectively analyzed. Among 31 AML children, their age were from 2-14 years old (median age 8 years old). There were 12,15 and 4 children classified in low risk group, intermediate risk group and high risk group by cytogenetic risk classification respectively. Eight children received concomitant hematopoietic stem cell transplantation. Kaplan Meier method with Log-Rank testing was employed for survival analysis.

Results: Follow-up was for a median 24 months (range: 5–50 months). The complete remission rate was 83.8%. The predicted 3-year leukemia free survival (LFS) rate was 53.8%. The LFS rate of low, intermediate and high risk group were 55.6%, 52.5% and 50.0% respectively. There was no significance in risk groups. The LFS rate of chemotherapy and chemotherapy concomitant HSCT were 42.7% and 87.5%, P<0.05. There were 2 cases of treatment related mortality including one case of sepsis and one case of ARDS.

Conclusions: NOPHO-AML 2004 study is clinically efficacious for the treatment of AML in Chinese children. HSCT treatment had better outcome than only chemotherapy in childhood with non low risk AML in CR1 phase.

Disclosures

No relevant conflicts of interest to declare.
Phase I-II Study of Sequential Therapy with Decitabine Followed By Clofarabine, Idarubicin, and Cytarabine (DAC-CIA regimen) in Relapsed/Refractory Acute Myeloid Leukemia (AML)
Authors: Jain Nitin , Ravandi Farhad , Garcia-Manero Guillermo , Borthakur Gautam , Kadia Tapan , Jabbour Elias , Pemmaraju Naveen , Daver Naval , DiNardo Courtney D. , Konopleva Marina , Pike Allison , Bivins Carol , Pierce Sherry , O'Brien Susan , Cortes Jorge E. , Kantarjian Hagop M.
Published: October 2019

Background: Patients (pts) with relapsed/refractory AML have dismal outcomes with currently available chemotherapy regimens. Preclinical and clinical evidence suggests that the use of hypomethylating agents such as decitabine prior to standard chemotherapy may be synergistic (Qin et al. Clin Can Res 2007; Scandura et al. Blood 2011; Clozel et al. Cancer Discov 2013).

Methods: Pts aged ≥18 to 65 years with relapsed or refractory AML (up to salvage 2) were eligible. Prior therapy with hypomethylating agents was allowed. The chemotherapy regimen was as follows: TableDoseDays INDUCTIONDecitabine20 mg/m2 IVDays 1-5Clofarabine15 mg/m2 IVDays 6-9 (Dose level 1) Days 6-10 (Dose level 2)Idarubicin10 mg/m2 IVDays 6-8Cytarabine1 gm/m2 IVDays 6-10CONSOLIDATIONDecitabine20 mg/m2 IVDays 1-5Clofarabine15 mg/m2 IVDays 6-8Idarubicin8 mg/m2 IVDays 6-7Cytarabine1 gm/m2 IVDays 6-8

Results: Eighteenpts (female, n=4) have been treated so far. Median age was 43 years (range 20-61). Majority of the pts (10/18, 56%) were in salvage 2. Four had a prior allogeneic stem cell transplant (SCT). Cytogenetics were diploid (n=4), complex (n=11), 11q abnormality (n=2), trisomy 8 (n=1). Five pts had a TP53 mutation and 3 pts had an NRAS mutation. Median number of cycles administered was 1 (range 1-2). Grade 3-4 non-hematological toxicity included grade 3 mucositis (DLT) in 2 pts during the induction cycle. Due to mucositis and prolonged cytopenias (prolonged cytopenias didn’t meet the DLT definition), dose level 2 was not pursued, and all patients were treated at dose level 1. Grade 3 transaminitis was seen in 4 pts. Fifteen pts had one or more infections. Six (33%) pts achieved CR, of whom 5 were able to proceed to SCT (one pt with CR opted not to pursue SCT). Median time to CR was 35 days. An additional 3 pts had a >50% decrease in bone marrow blast count (marrow response) and all three proceeded with SCT. All 4 patients with diploid cytogenetics achieved CR. Only 1/5 (20%) patients with mutated TP53 achieved a CR. Two patients have relapsed (one 3 months after SCT and the other after consolidation cycle 1). Six patients are alive (4 after stem cell transplant, 2 are receiving further salvage regimens after DAC-CIA failure). Median survival of the entire group is 7.2 months (see figure 1).

Conclusions: The sequential treatment of decitabine followed by chemotherapy is safe and effective with a CR rate of 33% and with 8/18 patients able to proceed to an allogeneic SCT. The phase II part of the study is enrolling patients at this time.

FIGURE 1 FIGURE 1.

Disclosures

Off Label Use: Decitabine and clofarabine are not approved for AML..
Diffusely Increased Bone Marrow 18F-FDG Uptake Is an Independent Predictor of Focal Bone Lesions in Patients with Newly Diagnosed Classical Hodgkin Lymphoma
Authors: Gormsen Lars C. , Wassberg Cecilia , Molin Daniel , Kamper Peter , Enblad Gunilla , d'Amore Francesco
Published: October 2019

Background: 18F-FDG PET/CT is recommended in the initial staging of patients with classical Hodgkin lymphoma (cHL). Whereas focal 18F-FDG uptake in the skeleton is considered to represent bone involvement, it is still unclear whether diffusely increased 18F-FDG bone marrow uptake (BMU) indicates lymphoma infiltration or merely reflects a state of general inflammation. This retrospective study was therefore performed to study the relationship between pre-therapeutic (PET0) 18F-FDG BMU and the presence of focal bone lesions.

Methods: A total of 139 patients (median age 44, range 8-83) referred to PET/CT between 2008 and 2014 for HL staging were included. All PET0 and post-therapeutic (PET2) images were reviewed and evidence of focal bone lesions was recorded (unifocal: ≤2 lesions, multifocal: >2 lesions). In addition, 18F-FDG uptake (SUVmax) was semiquantitatively measured in the vertebral bone marrow (SUVvertebra) and in the right lobe of the liver (SUVliver). BMU was calculated as SUVvertebra/SUVliver. The relationship between focal bone lesions on PET0 and BMU as well as age was subsequently analysed by logistic regression.

Results: In total 30/139 (22 %) patients had focal bone lesions at initial staging (10 unifocal, 20 multifocal). BMU at initial presentation was generally increased in all patients when compared with the post-therapeutic PET/CT (PET0: 1.22 +/- 0.03 vs. PET2: 0.95 +/- 0.03, p<0.001). In a logistic regression model diffusely increased BMU at PET0 (p=0.01) as well as age (p=0.01) were both independent predictors of focal bone lesions.

Conclusion: Diffusely increased 18F-FDG uptake is an independent predictor of skeletal bone lesions in patients with HL. Evidence of increased BMU might be evaluated as an additional outcome predictor in the pre-therapeutic risk assessment of patients with HL. With the purpose of validating these findings, we will analyze an independent cohort of Swedish cHL patients and joined data from both cohorts will be presented at the meeting.

Disclosures

No relevant conflicts of interest to declare.
BRG1 Expression in Diffuse Large B Cell Lymphoma (DLBCL) Is Associated with Advanced Disease Stage
Authors: Rehman Aseeb U , Nazeer Tipu , Wilner George D. , Laziuk Katsiaryna , Huho Albert N , Sheehan Christine S
Published: October 2019

Background: BRG1 is a component of the highly conserved multisubunit, 2.0-MDa mammalian transcriptional coactivator complex SWI/SNF. This chromatin-remodeling complex is critical for the regulation of gene transcription, thereby controlling diverse cellular processes including differentiation, proliferation, tumorigenesis and DNA repair. BRG1 gene is frequently down regulated or conversely, in some cases, up regulated in cancer cell lines and tumors, implicating that BRG1 may regulate cancer proliferation and progression. BRG1 is currently considered a potential selective therapeutic target. BRG1 expression level has been related to cancer cell migration and invasion and studies have suggested that BRG1 may serve as a prognostic marker in certain cancers. Few studies exist on expression of BRG1 in lymphoproliferative disorders and diffuse large B cell lymphoma (DLBCL) in particular. Our study was aimed at determining the immunohistochemical expression of the BRG1 protein in DLBCL and its potential association with clinicopathologic variables.

Design: Formalin-fixed, paraffin embedded sections from 49 Diffuse Large B-cell Lymphomas (DLBCL), were immunostained by a manual method (DAKO LSAB+ System-HRP) using rabbit monoclonal BRG1 (Abcam, Cambridge, MA) antibody. Nuclear and cytoplasmic immunoreactivity was semiquantitatively assessed in all cases. Scoring was based on staining intensity (weak, moderate, intense) and percentage of positive tumor cells (focal <= 10%, regional 11-50%, diffuse >50%). Results were correlated with clinicopathologic variables.

Results: BRG1 immunoreactivity was predominately nuclear. Intense diffuse nuclear BRG1 overexpression was observed in 22/49 (45%) cases and correlated with tumor stage [73% stage 4 vs 63% stage 3 vs 40% stage 2 vs 13% stage 1, (p=0.005)]. There were no other correlations that reached statistical significance.

Conclusion: BRG1 nuclear expression is increased in DLBCL and is associated with advanced tumor stage. This association with an important adverse prognostic indicator warrants further study of BRG1 expression and its potential as a selective therapeutic target in DLBCL.

Disclosures

No relevant conflicts of interest to declare.
Multiple Factors Analysis of B Cell Non-Hodgkin's Lymphoma Treatment-Related Interstitial Pneumonia
Authors: Zhao Jiali , Peng Yuna , Tan Huo
Published: October 2019

Objective To explore the related factors of interstitial pneumonia(IP) in B cell non-hodgkin's lymphoma (B - cNHL) patients who were treaded by chemotherapeutic drugs±RTX, which could provide certain references for clinical diagnosis and treatment .

Methods The clinical factors of 92 patients with B-cNHL enrolled from 2003 to 2013 in my hospitol were retrospectively studied, then cleared the high risk factors influencing the occurrence of IP.

Results The statistical analysis found that combined application of chemotherapeutic drugs and RTX was the only independent risk factor of IP,which have statistical Significance (OR=9.6; p<0.05) .While the other clinical factors, such as age, sex, past history of chronic lung disease and smoking, tumor’s clinical stage, International Prognostic Index (IPI) score, Eastern Cooperative Oncology Group-performance status(ECOG PS) score, complete blood cell count, LDH, bone marrowd and pulmonary parenchyma had nothing to do with the happening of the IP.Finally, compared chemotherapeutic drugs+RTX group and chemotherapeutic drugs group by survival analysis, the incidence of IP of the former was higher than the latter, and had statistical significance (P = 0.011).

Conclusion The incidence of IP of chemotherapeutic drugs+RTX group was obviously higher than chemotherapeutic drugs group. Addition of RTX to chemotherapy was identified as an independent risk factor for B-cNHL treatment-related IP. Once patients suffering from IP should Clinicians carefully identify IP caused by opportunistic pathogens and DIIP caused by RTX.

Disclosures

No relevant conflicts of interest to declare.
Coexpression of MYC and BCL-2 Predicts Inferior Survival in Pgi-DLBCL Treated with CHOP-like Regimen/Rituximab
Authors: Zhang Le , Xia Bing , Guo Shanqi , Li Xiaowu , Qu Fulian , Zhao Haifeng , Zhang Lianyu , Sun Baocun , Zhang Liang , Zhang Yizhuo
Published: October 2019

MYC protein expression has been identified to be associated with inferior overall survival (OS) and progression-free survival (PFS) when coexpressed with BCL-2 protein in patients with diffuse large B cell lymphoma (DLBCL). But the concurrent expression of MYC and BCL-2 proteins in primary gastrointestinal (PGI)-DLBCL has not been clearly understood. Here, we investigated whether this coexpression has prognostic significance in PGI-DLBCL patients and explored its associations with patients’ clinical parameters. We enrolled 60 PGI-DLBCL patients and 30 age- and sex-matched healthy controls. Expression levels of MYC and BCL-2 were detected from both protein and mRNA levels by immunohistochemistry and real-time RT-PCR. Positive expression levels of MYC and BCL-2 proteins were detected in 35% and 45% of patients, respectively. MYC+/BCL-2+ protein was present in 30% of patients. MYC and BCL-2 protein were correlated with high MYC and BCL-2 mRNA expression, respectively (both p<0.05). We found that patients with advanced-stage disease (at IIE-IV) having higher MYC and BCL-2 coexpression levels (p<0.05). In addition, MYC+/BCL-2+ patients had more difficulty achieving complete remission than others (p<0.05). Presence of MYC protein expression only affected OS and PFS when BCL-2 protein was coexpressed. The adverse prognostic impact of MYC+/BCL-2+ protein on PFS remained significant (p<0.05) even after adjusting for age, Lugano stage, IPI, and BCL-2 protein expression in a multivariable model. MYC+/BCL-2+ patients have poorer chemotherapy response and poorer prognosis than patients who only express one of the two proteins, suggesting that assessment of MYC and BCL-2 expression by immunohistochemistry has clinical significance in predicting prognosis of PGI-DLBCL patients.

Disclosures

No relevant conflicts of interest to declare.
Transdifferentiation of Malignant B-Cells into Macrophages in a Murine Model of Burkitt´s Lymphoma
Authors: Bruns Heiko , Mueller Sebastian , Bach Christian , Pasemann Shirin , Maurberger Anna , Mackensen Andreas , Gerbitz Armin
Published: October 2019

B-cell lymphomas, such as Burkitt's lymphoma, are malignant diseases of the hematopoietic system. They arise from transformed B-cells originating from diverse primary and secondary lymphatic tissues. Like in other cancers, the stroma of lymphomas is typically infiltrated by so-called tumor-associated macrophages (TAMs), which execute manifold tumor-specific functions. Interestingly it was shown, that mature B-cells could be efficiently re-programmed into macrophages by the overexpression of myeloid-specific transcription factors. Moreover, other studies observed in vitro that this lymphoid/myeloid plasticity might be also caused by oncogenes in cultured B-cells of murine lymphoma models. Therefore we consider, if lymphoma B-cells themselves might be a source of TAMs, besides the well-known infiltration of monocytic cells into the tumor environment. Based on the system of CD45.1/2 allotypes, a murine model of lymphoma was thus developed, which allows tracking of the conversion of lymphoma B-cells into TAMs.

It could be shown, that some lymphoma B-cells of the established model in fact spontaneously switched into a macrophage-like phenotype. Accordingly, they start to express typical macrophage markers, but seem to perform a transition into a myeloid-like expression profile on the level of transcription as well. Furthermore, analysis of the recombined immunoglobulin heavy chain confirmed the clonal identity of lymphoma b-cells and TAMs. Although these cells do not exhibit an overt immunological phenotype, they show elementary functional properties of macrophages, such as phagocytosis as well as a post-mitotic character. Moreover, transdifferentiated B-cells cells were resistant to chemotherapy and persist after treatment, while lymphoma B-cells were eradicated.

Which consequences the lymphoid/myeloid plasticity of lymphoma B-cells has for tumor development, progression, as well as for the outcome of therapy, and if these observations could be transferred to human tumors, will be object of future investigations.

Disclosures

No relevant conflicts of interest to declare.
Adverse Prognostic Impact of Neutrophil-to-Lymphocyte Ratio and Platelet-to-Lymphocyte Ratio in Patients with Diffuse Large B Cell Lymphoma
Authors: Kong Sun-Young , Lee Hyewon , Sohn Ji Yeon , Eom Hyeon-Seok
Published: October 2019

Neutrophil-lymphocyte ratio (NLR) and platelet-to-lymphocyte ratio (PLR) have been lightened as a prognostic factor in many types of solid tumor. However, there was a limited report for diffuse large B cell lymphoma (DLBCL). Thus, we investigated the impact of NLR and PLR on prognosis of patients with DLBCL. This study involved 234 DLBCL patients treated by rituximab plus cyclophosphamide, doxorubicin, vincristine, and prednisolone (R-CHOP) at National Cancer Center, Korea. The median patient age was 57 years, and 135 patients (58%) were men. Clinical characteristics as international prognostic index (IPI) including age, performance, LDH, stage and extra-nodal involvement, were evaluated. NLR and PLR at diagnosis were calculated by CBC data from automated hematologic analyzer XE2100 (Sysmex, Kobe, Japan) or differential count which was done by manually using stained peripheral blood slide when CBC data showed flags in auto-analyzer. Hazard ratios (HRs) were determined in terms of risk for overall survival using Cox proportional hazard regression analysis. The mean percentage of neutrophils and lymphocytes was 60 (range, 5-95) and 12 (2-79), and the number of platelets was 257 × 10^9/L (28-839). The mean and standard deviation of NLR and PLR was 3.62 (±4.77) and 14.7 (±20.16), respectively. IPI and stage were predictors for prognosis (p <0.01), and both NLR and PLR showed significant association with poor prognosis of HRs of 1.14 [95% confidence interval (CI), 1.10 to 1.20] and 1.02 [95%CI, 1.02-1.03]. Although both NLR and PLR represented association with IPI, there were still prognostic impacts as HR of 1.12 and 1.02 with IPI in multivariate analysis. These findings suggest that NRL and PLR may be significant prognostic indicators for patients with DLBCL treated by R-CHOP.

Disclosures

No relevant conflicts of interest to declare.
Development of Non-Hodgkin’s Lymphoma As Second Primary Cancer in Hepatitis C Virus-Infected Patients with a Different Primary Malignancy – a Case Series
Authors: Mahale Parag , Romaguera Jorge , Turturro Francesco , Fowler Nathan , Torres Harrys A.
Published: October 2019

Background: Hepatitis C virus (HCV) is carcinogenic and infected patients are at increased risk of developing hepatocellular carcinoma (HCC) and non-Hodgkin’s lymphoma (NHL). HCC can develop as a as a second primary cancer (SPC) in HCV-infected cancer patients with a nonliver first primary cancer. However, reports on HCV-infected patients who develop NHL as SPC are lacking.

Methods: Records of patients with chronic HCV infection who were seen at MD Anderson Cancer Center, Houston, TX, between 01/2008 - 5/2014 were reviewed. Patients who developed NHL in the setting of a different primary malignancy were described in this case series. HCV therapy did not include newly approved antivirals.

Results: Eighty four patients with HCV-associated NHL were seen during the study period. Among, them, six (7%) developed NHL as SPC; most were men (n=4; 67%), Caucasians (n=5; 83%), and had genotype 1 infection (two patients had genotype known; both had genotype 1b infection). All patients had solid first primary cancers, mostly prostate cancer (n=3; 50%), followed by choriocarcinoma (n=1; 17%), chondrosarcoma (n=1; 17%), and squamous cell carcinoma of larynx (n=1; 17%). Most patients (n=5; 83%) had their first primary cancers in complete remission, and only 1 had cirrhosis at lymphoma diagnosis. Surgery (n=3; 50%) and chemotherapy (n=2; 34%) were the main treatment modalities for first primary malignancies, with leuprolide acetate and carboplatin as the only chemotherapeutic agents administered. The types of NHL developed as SPC were diffuse large B cell lymphoma (n=3; 50%), marginal zone B cell lymphoma (n=2; 33%), and mantle cell lymphoma (n=1; 17%). None of the patients had a family history of lymphoma. NHL was not suspected and was found incidentally in all patients at a median of 34 months (range, 2 – 410 months) after the diagnosis of the first primary malignancy. Hepatitis B core antibodies were detected in 4 (67%) patients. A majority of patients had persistent HCV viremia (n=4; 67%) when NHL was diagnosed. Two of the 3 patients who were treated for HCV infection had attained sustained virological response (regarded as virological cure).

Conclusions: NHL may develop as SPC in HCV-infected patients with a different primary malignancy, especially in patients who have primary solid tumors or unresolved infection. Therefore, surveillance not only for the development of HCC but also NHL as SPCs should be continued in HCV-infected patients with different primary malignancy. Eradication of this carcinogenic virus with better tolerated and more efficacious antivirals may potentially reduce the risk of NHL development as SPC.

Disclosures

Torres: Gilead Sciences: Consultancy; Merck & Co., Inc. : Consultancy, Research Funding; Vertex Pharmaceuticals: Consultancy, Research Funding; Genentech,: Consultancy.
Bleomycin Pulmonary Toxicity in Adult Saudi Patients with Hodgkin’s Lymphoma
Authors: Al-Mansour Mubarak M , Al-jizani Wafa A , Al-fayea Turki , Ruaa Shafi , Kazkaz Ghieth A , Bayer Ali M , AL-Foheidi Meteb , Ibrahim Ezzeldin M
Published: October 2019

Background: Bleomycin pulmonary toxicity (BPT) has been well described in Hodgkin's lymphoma (HL) patients treated with bleomycin-containing chemotherapy regimens. As the effects of genetic susceptibility or race on the development of BPT are largely unknown, we intended to examine BPT incidence, risk factors, and its effects on survival in a series of newly diagnosed Saudi Arabian adult HL patients receiving first-line bleomycin-containing chemotherapy.

Methods: We carried out a retrospective review of the relevant records of 164 consecutive eligible adult HL patients treated at two centers.

Results: BPT was observed in 24 of 164 patients (15%) at a median duration of 4.3 months (range, 1 to 58 months). Older age and history of concomitant lung disease were significantly associated with approximately three- (odds ratio [OR] = 3.38; 95% confidence interval [CI], 1.25-9.13; P = 0.02) and seven-fold (OR = 7.19; 95% CI, 2.64-19.54; P <0.0001) increase in BPT risk, respectively. While more patients in non-BPT group achieved CR as compared with BPT patients (93% vs. 79%; P = 0.048), the actuarial 5-year progression-free (82% vs. 63%; P = 0.29) and overall survival (OS) (94% vs. 76%; P = 0.059) for BPT and non-BPT groups, respectively were not significantly different. Nevertheless, in a multivariate analysis, the earlier incidence time of BPT considered as a time-dependent covariate was the only factor that independently influenced OS (hazard ratio = 0.99; 95% CI, 0.99-1.00; P = 0.002).

Conclusion: In Saudi Arabian adult HL patients, the risk of BPT and its effect on survival outcome were comparable to that reported from developed countries. Earlier incidence time of BPT aversely influenced OS. Future studies should consider testing the efficacy and safety of non-bleomycin-containing regimens in HL.

Disclosures

No relevant conflicts of interest to declare.
Generic Imatinib in a Patients with Chronic Myeloid Leukemia in Chronic Phase- a Single Centre Experience
Authors: Urosevic Ivana M , Dokic Marina , Savic Ivanka , Savic Aleksandar , Milosevic Ivana , Rajic Nebojsa , Andjelkovic Nebojsa V. , Sekulic Borivoj , Vlaisavljevic Nada V
Published: October 2019

Imatinib (Gleevec) has shown long-term efficancy and safety in published randomized clinical trials. Anzovip (Imatinib) is a new generic drug produced by Zdravlje Actavis,which was approved by the Medicines and Medical Devices Agency of Serbia in January 2012. In July 2012, National Health Insurance Fund of Serbia introduced this drug in the positive list. All the patients using Glivec were switched to Anzovip and all newly diagnosed patients started with Anzovip therapy. In this study, hematologic and cytogenetic responses were evaluated in CML patients who received generic Imatinib as the first line therapy and in patients who switched from branded Imatinib to an Imatinib copy drug.

Materials and methods. During August and September 2012, all the patients were switched from Glivec to Anzovip and all new CML patients were treated with Imatinib copy. 55 patients treated with branded Imatinib were switched to generic Imatinib. Out of 17 newly diagnosed patients with CML-CP, who started with generic Imatinib, 7 patients could be followed up for 18 months. All patients underwent hematologic, cytogenetic monitoring based of the recommendations of the ELN.

Results. These are the results of using Anzovip after 18 months. Of the 55 patients who were switched from branded Imatinib to Imatinib copy, 11 patients (20 %) have lost complete cytogenetic response they already had, but without signs of biological illness transformation. They were switched to 2nd line therapy nilotinib.Of the 7 newly diagnosed patients who started with genereic Imatinib, all 7 patients (100%) have achieved complete hematologic response (CHR) in the period of 6 months. 3 (42,8%) patients have achieved CCgR and 2 patients (28,6%) have achieved MMR in the period of 12 months. 57% of patients have switched to second generation of TKI because of failure or intolerance according to ELN and national recommendation. The drug has been well tolerated, wild mild adverse event, more frequently, fluid retention and gastrointestinal symptoms. Only one patient have had skin changes grade ¾ and he has been switched to second generation of TKI.

Conclusions. It is unknown whether patients, who responded to branded Imatinib and than switched to its copy versions, will tolerate the copy drug and maintain the previous response. Because of that, it is important to careful follow up of a selected patients several months after the switch to generic imatinib. Despite of the small number of patients who started with Imatinib copy, our results in term of hematologic and cytogenetic response were close to the international series.

Disclosures

Urosevic: Novartis: Consultancy, Honoraria. Savic:Roche: Honoraria; NovoNordisc: Honoraria. Rajic:NovoNordisc: Honoraria.
Mantle Cell Lymphoma, Rare Lymphoma Entity: A Result of Lymphoma Registry in Thailand
Authors: Kanitsap Nonglak , Wannissorn Naree , Lekhakula Arnuparp , Julamanee Jakrawadee , Khuhapinant Archrob , Sirijerachai Chittima , Chansung Kanchana , Norasetthada Lalita , Nawarawong Weerasak , Chuncharunee Suporn , Niparuck Pimjai , Wudhikarn Kitsada , Bunworasate Udomsak , Numbenjapon Tontanai , Prayongratana Kannadit , Wongkhantee Somchai , Wong Peerapon , Suwanban Tawatchai , Makruasi Nisa , Intragumtornchai Tanin
Published: October 2019

Introduction: Mantle cell lymphoma (MCL) is a relatively rare and aggressive mature B-cell non-Hodgkin lymphoma (NHL) entity, encounting for 3-10% of newly diagnosed patients. The purpose of the study is to analyse the incidence, clinical features and outcomes of Thai patients with MCL and compare data with those of Western countries.

Patients and methods: From the nationwide multicenter registry of 4,056 newly diagnosed NHL patients in Thailand between 2007-2014, patients with MCL were identified according to the criteria of diagnosis by WHO classification of lymphoid disorders, 2008. The incidence, baseline clinical characteristics and outcomes were then analysed.

Results: Ninety-nine patients (2.4%) were found to have MCL. The median age was 59.8 years (range 33-86). Seventy-nine percent of patients were men. The primary sites of disease were lymph nodes (60%), bone marrow (19%), GI tract (9%), Waldeyer's ring (4%), sinonasal area (2%) and miscellaneous (6%). (Figure 1). Eighty-three percent of the patients had stage III/IV. Eighty-one patients (82%) received chemotherapy (60.5% with CHOP or CHOP-like chemotherapy, 30.9% with HyperCVAD, and 26% with rituximab-based). Other received palliative chemotherapy or did not receive any. At a median follow-up time of 42 months, the median survival was 30 months (range, 1-94 months). By using Cox analysis, only Ann Arbor stage had impact on overall survival.

Conclusion: MCL is less frequent in Thailand compare to the Western countries. The long-term survival is also much inferior reflecting the limited access to rituximab as well as the high intensity therapy in treating MCL in Thailand.

Table 1. Comparison of MCL patients according to treatment modalities Clinical characteristics (Total 81 cases) CHOP-like (n=49 ) High intensity CMT (n = 25 ) CVP or palliative CMT (n = 7 ) p-value No R 33 19 4 - Age > 60 19 (67%) 6 (24%) 5 (71%) - Stage III-IV 42 (85.7%) 21 (84%) 7 (100%) - Response Overall response Complete response 26 (53.1%) 19 (38.8%) 17 (68%) 11 (44%) 6 (85.7%) 3 (42.8%) - Survival Mean OS (months) (Min-Max) 3yr-OS 5yr-OS 26.8 (0.1-83) 52.2% 52.2% 38.8 (0.8-93.9) 51% 38.1% 34.3 (10.7-70.5) 51% 30.2% 0.44 0.44

Figure 1. Biopsy sites Figure 1. Biopsy sites Figure 2. The Kaplan-Meier plots of OS according to treatment groups Figure 2. The Kaplan-Meier plots of OS according to treatment groups Figure 3. The Kaplan-Meier plots of OS according to tretment with rituximab Figure 3. The Kaplan-Meier plots of OS according to tretment with rituximab

Disclosures

Khuhapinant: Roche: Honoraria.
Investigation of Two Combination HbF Induction Regimens, RN-1 and Hydroxyurea Versus RN-1 and Decitabine, in a Humanized Sickle Cell Mouse Model
Authors: Jagadeeswaran Ramasamy , Agarwal Yash , Ibanez Vinzon , Ruiz Maria Armila , Vaitkus Kestis , DeSimone Joseph , Lavelle Donald , Rivers Angela
Published: October 2019

Sickle cell disease (SCD) results from the substitution of glutamic acid for valine at the sixth amino acid of β-globin protein, leading to the formation of the abnormal hemoglobin S (HbS). Following deoxygenation in red blood cells (RBCs), HbS forms polymers causing the RBCs to become deformed and adherent leading to vaso-occlusive events resulting in splenic infarction, kidney failure, stroke, painful crises, chronic anemia, inflammation and decreased survival. Hydroxyurea, the only drug currently FDA approved for SCD, is not effective in some patients and, therefore, new combination regimens are needed. RN-1, a new drug targeting the histone demethylase LSD-1, is a more effective inducer of γ-globin expression in sickle cell mice and HbF in baboons (P. anubis). Decitabine(DAC), a DNMT1 inhibitor, is well known to induce high levels of HbF in non-human primates and SCD patients and is currently in clinical trials. We investigated the effect of two combination regimens: RN-1 in combination with hydroxyurea vs. RN-1 in combination with DAC. The knock-in humanized sickle cell mice (B6;129-Hbatm1(HBA)Tow Hbbtm2(HBG1,HBB*)Tow/J) were treated with 1) DMSO (vehicle control), 2) RN-1 (5mg/kg), 3) DAC(0.25mg/kg) and 4) RN-1(5mg/kg) + DAC(0.5mg/kg) for 10 days. Analysis of F-retics and γ-globin mRNA performed on d10 showed F-retics were increased two fold (p=0.01) in mice treated with RN-1(5mg/kg) or DAC(0.25mg/kg) compared to DMSO-treated. γ-globin mRNA levels (γ/γ+β) were increased three fold in RN-1(5mg/kg) or DAC(0.25mg/kg) compared to DMSO. The combination of RN-1(5mg/kg) and DAC(0.25mg/kg) did increase F-retics and mRNA levels compared to mice treated with RN-1 alone(p=0.02). Also, the RN-1 and DAC combination led to neutropenia and thrombocytopenia. We next examined the effect of RN-1(5mg/kg) and HU (100mg/kg) in combination. Knock-in humanized sickle cell mice (B6;129-Hbatm1(HBA)Tow Hbbtm2(HBG1,HBB)Tow/J) were treated with 1) DMSO (vehicle) 2) RN-1(5mg/kg) and 3) RN-1(5mg/kg) + HU (100mg/kg). While RN-1 in combination with HU did not significantly increase F-cells or F-retics, there was a significant decrease in spleen weight [(DMSO: 1.17±0..06g; RN1:0.71± 0.1g; RN1+HU: 0.46±0.05g)(p=0.004:p=0.02)] and percentage of sickle cells in a peripheral blood smear.[(DMSO: 8.4 ±1.3%; RN-1:4.1± 0.2%; RN1+HU: 1.57±0.6%)(p=0.006:p-0.014)]. We conclude that further studies to determine the effect of RN-1 in combination with HU or DAC in the sickle cell mouse model and cultures from sickle cell CD34+ cells are needed to evaluate its potential use in the treatment of sickle cell disease. These results suggest that RN-1 in combination with hydroxyurea may be a useful therapeutic regimen.

Disclosures

No relevant conflicts of interest to declare.
Effect of PSI697, a Small Molecule Inhibitor of P-Selectin, in the Townes Model of Sickle Cell Disease
Authors: Jasuja Reema , Hett Sunita Patel , Kaila Neelu , Pittman Debra
Published: October 2019

Chronic vaso-occlusion is a major cause of morbidity and mortality in patients with sickle cell disease (SCD). Persistent vaso-occlusion can damage lungs, liver, kidneys or brain and ultimately lead to end-organ dysfunction. Vaso-occlusive pain crisis is a complex multistep process, initiated by adhesion of fragile sickle erythrocytes and rigid neutrophils to hypoxic and inflamed endothelium. Large multicellular aggregates of blood cells, including platelets and sickled erythrocytes, form on these adherent activated neutrophils in the microcirculation, ultimately causing vascular occlusion and tissue ischemia. Inflammatory mediators, such as P-selectin, play a key role in mediating these heterocellular interactions and attract additional leucocytes to the site of occlusion. P- and E-selectin mediate rolling and tethering of blood cells on the endothelium. Here, we explore cellular and animal models of sickle cell disease to assess PSI-697, an orally active small molecule antagonist of P-selectin. PSI-697 inhibits P-selectin binding to P-selectin glycoprotein ligand-1 (PSGL-1) with an IC50 of 50-125 µM (Bedard et al, JPET, 2008). Townes SCD mice were used to assess the efficacy for PSI-697 in preventing vaso-occlusion in SCD. Mice were randomized to treatment with vehicle or PSI-697. Animals were treated in a prophylactic setting prior to the surgical preparation. Surgical preparation induces a well described acute inflammatory response in the microcirculation associated with neutrophil adhesion to the endothelium and formation of multicellular aggregates. Alexa-488 labeled Ly-6G neutrophil antibody and Dylight-649 labeled CD42c platelet antibody were injected to quantify adherent neutrophils and neutrophil-platelet aggregates. Cremaster microvasculature was observed by intravital microscopy. Mice treated with 100 mg/kg of P-selectin inhibitor PSI-697 showed a 55% reduction in adherent neutrophils and a 78% decrease in the number of neutrophil-platelet aggregates compared to vehicle treated animals. Neutrophils showed ~7 fold increase in rolling velocity in mice treated with P-selectin inhibitor. Our results demonstrate that prophylactic administration of PSI-697, a small molecule P-selectin antagonist, improved parameters associated with vaso-occlusion in Townes SCD mice.

Disclosures

No relevant conflicts of interest to declare.
Pain Experiences in Children with Sickle Cell Disease in the Era of Hydroxyurea
Authors: Kavanagh Patricia , Sprinz Philippa G. , Libernays Evelyn , Pavlus Nicole , Wang C. Jason
Published: October 2019

Background: Pain is the most common complication of sickle cell disease (SCD), and pain diaries are an effective way to capture pain experienced by both children and adults for many conditions, including SCD. The most recent data on pain experienced by children with SCD outside the clinical trial setting, particularly those with Hemoglobin (Hb) SS disease, were collected prior to the publication of the Baby-HUG trial in 2011, which found hydroxyurea to be safe for use in early childhood. One study found that school-aged children with SCD had at least one pain episode per month; another longitudinal study reported that SCD-related pain was experienced by children on 8% of pain diary days. In the era of hydroxyurea use in children, the occurrence of SCD-related pain episodes in children has not been well studied, especially among those with Hb SS disease.

Methods: Parent/child dyads were recruited for a 6-month pilot study of the use of home pain plans and pain diaries from a pediatric hematology clinic based in a safety net institution. The study ran from February to November 2014. Dyads were eligible if the child with SCD was aged 5-15 years and had at least one episode of pain in the year prior to enrollment, by parent report. Parents and children were asked to fill out the diary each day. They received weekly phone calls for the first month, then monthly phone calls until the close of the study, to remind them to complete the daily pain diary and mail it back monthly; $20 gift cards were given for each diary returned. We excluded data from dyads that recorded less than 4 weeks of pain diary data.

Results: Thirty-six parent/child dyads were approached and 30 (83%) consented to enroll in the study. Twenty-six (87%) dyads recorded at least four weeks of data. The average age of the child with SCD was 10.3 ± 3.0 years and 20 (77%) were insured through Medicaid. Twenty-two (85%) children had Hb SS disease, of whom 21 were prescribed hydroxyurea. Data were recorded per dyad on average 135 ± 46 diary days. One child with Hb SS disease reported no pain during the study period. The remaining 25 children had any pain and SCD-related pain an average of 8% and 4% diary days, respectively. Nearly all children reporting non-SCD-related pain had headache or abdominal pain. Ten (38%) children missed at least one day of school during the study period.

Conclusions: In thissample of children with SCD, composed predominantly of those with Hb SS disease prescribed hydroxyurea, pain was an infrequent occurrence and was attributed to SCD only half the time. This study suggests that SCD-related pain experienced by children may be lower than previously reported. Further study is warranted, especially among those prescribed hydroxyurea.

Disclosures

No relevant conflicts of interest to declare.
Pharmacogenocis of PAR4: PAR4 Polymorphism Determines Platelet Response in the Presence of Dual Anti-Platelet Therapy
Authors: Tourdot Benjamin , Bray Paul F. , Edelstein Leonard C. , Holinstat Michael
Published: October 2019

Aberrant platelet activation contributes to the mortality associated with coronary heart disease (CHD). Our lab group has recently shown that platelets from black donors were hyper-responsive to activation of the thrombin receptor Protease-Activated Receptor-4 (PAR4) compared to platelets from white donors. The racial difference observed through PAR4 persists in platelets treated ex vivo with dual antiplatelet inhibitors (COX-1 and P2Y12 receptor inhibitors), a standard of care for many CHD patients. The racial difference in PAR4 signaling is largely attributable to one PAR4 variant more frequently expressed in blacks than whites, PAR4 T120A. Independent of race, T120 is associated with greater PAR4-mediated platelet reactivity compared to the A120 variant. The role PAR4 variation plays in regulating thrombin-mediated platelet activation in CHD has not been elucidated to date. We hypothesize that people homozygous for the T120 will have an increase in PAR4-mediated platelet reactivity compared to patients homozygous for A120. To investigate this hypothesis, we studied PAR4 signaling in platelets obtained from healthy donors. Platelet activity was assessed in each subject at the beginning of the study. Subsequently subjects were placed on a 7-day treatment of either COX-1 or P2Y12 inhibitors and on day 7 their blood was again drawn and tested for PAR4-AP-mediated platelet activation. Subjects who are homozygous for T120, have an increase in platelet reactivity as measured by aggregation, αIIbβ3 activation and granule secretion compared to individuals who are homozygous for A120. Further distal signaling nodes such as calcium and Rap1 are hyperactive in individuals who are homozygous for T120. This study reinforces the personalized medicine approach to therapeutic intervention and challenges the one size fits all approach which often leaves at risk populations without adequate protection from thrombotic events and stroke. This study was supported in part by grants MD007880, GM105671, and HL 114405 (MH) from the NIH.

Disclosures

No relevant conflicts of interest to declare.