bluuebunny/crossref_metadata_2025 huggingface dataset:
Glycoprotein Ibα Clustering Induces Macrophage-Mediated Platelet Clearance in the Liver
Authors: Yan Rong , Chen Mengxing , Ma Na , Zhao Lili , Cao Lijuan , Zhang Yiwen , Zhang Jie , Yu Ziqiang , Wang Zhaoyue , Xia Lijun , Ruan Changgeng , Dai Kesheng
Published: October 2019

Objective: Immune thrombocytopenia (ITP) is an autoimmune bleeding disease in which autoantibodies are directed against the patient’s own platelets, leading to platelet destruction and bleeding. Currently, platelet destruction in ITP is thought to be mediated by monocytic and phagocytic cells, and the antibody-coated platelets are removed from circulation by the spleen in an Fc-dependent manner. Conventional treatments for ITP therefore aim at reducing platelet destruction, either by immunosuppression or splenectomy. However, many ITP patients, particularly patients with anti-glycoprotein (GP) Ib-IX autoantibodies, do not respond to conventional treatments such as intravenous immunoglobulin G (IVIG). However, the underlying mechanism remains unclear.

Methods: Anti-GPIb-IX antibodies-induced platelet aggregation was detected by an optical aggregometer. GPIbα clustering was measured by Fluorescence Resonance Energy Transfer (FRET) by flow cytometry. Antibodies-induced platelet depletion and rescue experiments were performed in guinea pigs and cynomolgus macaques. Platelet distribution after antibody-mediated depletion was detected by in vivo Imaging System (IVIS), immunohistochemistry and immunofluorescence microscopy.

Results: We found that anti-GPIbα N-terminus antibody AN51, but not other anti-GPIbα antibodies (AK2, HIP1, VM16d, or WM23), induced integrin αⅡbβ3-dependent platelet aggregation. FRET result showed that AN51 induced obvious GPIbα clustering but not VM16d. After intravenous injection, AN51 dose-dependently induced thrombocytopenia in guinea pigs, and the platelets were mainly removed by macrophages in the liver. N-acetyl-D-glucosamine (GlcNAc), previously shown to inhibited αMβ2-mediated phagocytosis of refrigerated platelets, showed dose-dependent inhibition of AN51-induced platelet destruction. Furthermore, AN51 but not VM16d, induced rapid platelet clearance in the liver of cynomolgus macaques. Five of twenty-two chronic ITP patients had anti-GPIbα autoantibodies, and the autoantibodies from four of the five patients competed with AN51 for binding to platelets.

Conclusion: GPIbα clustering induced by anti-GPIbα N-terminus antibody causes integrin αⅡbβ3-dependent platelet aggregation, phagocytosis, and rapid platelet clearance in the liver. Our findings reveal a novel Fc-independent mechanism underlying the pathogenesis of ITP, and suggest new therapeutic strategies for ITP patients with anti-GPIbα autoantibodies.

Disclosures

No relevant conflicts of interest to declare.
Feasibility of Population Scale Comprehensive Identification and Analysis of Complex Structural Variations in Cancer Genome Using Nanochannel Array
Authors: Hastie Alex , Lam Ernest , Dai Heng , Andrews Warren , Pang Andy , Anantharaman Thomas , Saghbini Michael , Sadoski Henry , Austin Michael , Dickinson Todd , Yang Xing , Dzakula Zeljko , Rossi Michael R. , Kwok Pui , Xu Xun , Cao Han
Published: October 2019

Diseases with complex traits such as hematological cancer are known to be associated with large structural variations (SV > 1 kb). Techniques such as karyotyping, FISH and aCGH have been common tools for gross chromosomal lesion analysis. Yet population scale comprehensive SV analysis with these tools remains impractical, tedious or incomplete such as missing balanced lesions by aCGH. Recently, high throughput Next Generation Sequencing (NGS) has generated a large amount of sequencing reads that have rapidly reduced costs and are effective in detection of SNPs and small indels; however, complicated by the fact that large structural variations, often spanning tens to hundreds of thousands of base pairs or involving complex rearrangements throughout the genome, are hard for current short read sequencing technology to assemble or infer. Therefore, there is a blind spot in effectively detecting SVs within this range (1 kb ~ 1 Mb) due to insufficient tools.

Here we demonstrate a technology that rapidly linearizes very long strands of genomic DNA (100 kb to Mbs) through NanoChannels to directly visualize large SVs and rearrangements preserved within intact genomic DNA at the single molecule level. Through specific sequence motif labeling, de novo genome physical maps are assembled within hours and hundreds to thousands of SV events, balanced or imbalanced, are called with no a priori knowledge of the samples. Preliminary testing data from a group of cancer samples and multiple trio families will be shown to demonstrate this highly comprehensive and cost effective approach, with results validated by direct single-molecule images and multiple orthogonal methods. We also show long spanning molecules would provide very valuable information of precise mapping of viral genomic sequence integration in human genomes that are potentially associated with malignant cell transformation. For the first time, it is now feasible to do large population-based comprehensive genome structural variation studies on a single platform. This innovations will transform the biomedical research, diagnosis and treatment of hematological cancers that result from structural variations and chromosomal lesions.

Disclosures

No relevant conflicts of interest to declare.
Meta-Analysis of Pharmacotherapy Vs. Observation for Management of Smoldering Multiple Myeloma
Authors: Vijayakumar Jayanthi , Jackson Jeffrey , Ahn Kwang W , Hari Parameswaran N.
Published: October 2019

Introduction:

Asymptomatic or smoldering multiple myeloma (SMM) is usually 10-15% of all patients with multiple myeloma. Currently, national guidelines do not recommend early treatment of SMM though the majority of patients progress to active disease over time. Multiple trials have been conducted to evaluate implications of early therapy for SMM. Some of these trials, most notably, Mateos et al. (N Engl J Med 2013; 369:438-447) have shown benefit for treating SMM. However, there is lack of consensus as to whether early treatment of SMM prior to symptomatic disease progression is superior to observation alone. Hence we conducted a meta- analysis to determine if observation or treatment is superior in the management of SMM.

Methods:

We searched MEDLINE, the Cochrane database and hand-search of articles to identify 12 clinical trials which allocated SMM patients to treatment. Study quality was assessed using the Cochrane risk of Bias tool. Our main outcomes were progression free survival (PFS) and overall survival (OS). Sub-analysis examined the difference in treatment with antineoplastic agent (C), bisphosphonates (B), or their combination (CB). Data were pooled using a random effects meta-analysis using STATA (College Station Texas, V13.2).

Results:

Among the 12 trials, 4 trials were excluded due lack of survival curves for OS, PFS, TTP. Characteristics of the 8 included clinical trials are given in Table 1. Participants treated for early SMM had reduced mortality (HR: 0.64, 95% CI 0.4-1.0) and better PFS (HR=0.83, 95% CI, 0.64-1.07) compared to the observation group. In subgroup analysis, patients treated with either antineoplastic agents or the combination of antineoplastic agents and bisphosphonate had a better OS and PFS, whereas therapy with only bisphosphonate did not impact OS or PFS (Figures 1, 2).

Conclusion:

We conclude that antineoplastic treatment or a combination of antineoplastic agents and bisphosphonates is better than observation in improving overall survival in SMM patients. A similar trend is seen with PFS. This conclusion merits caution given the limited data available and the differences in the agents used for therapy.

Figure 1 Figure 1.

Figure 2 Figure 2.

Disclosures

No relevant conflicts of interest to declare.
A Comparison of Human CMVpp65-Specific Central Memory and Effector Memory CD8 T-Cells When Stimulated in-Vivo with IL-2 or IL-15/IL-15Rα Complex in a Murine Xenograft Model of Adoptive Cell Therapy
Authors: Kuo Tzu-Yun , Hasan Aisha , O'Reilly Richard J
Published: October 2019

Initial clinical trials of adoptive immunotherapy have shown that the efficacy of adoptively transferred T-cells in man is often limited by the failure of cultured T cells, particularly cloned CD8 T cells, to persist in vivo. These studies demonstrated that the transferred T cells induced only transient responses and that persistence of the transferred T-cell clonotypes correlated with disease regression. A previous study suggested that CMV virus-specific CD8 T cell clones derived from central memory T cells (TCM), but not effector memory T cells (TEM), persisted long-term in non-human primates. On the other hand, another study comparing TCM and TEM derived SIV virus specific CD8 T-cell clones that were adoptively transferred in non-human primates demonstrated limited persistence of both TCM and TEM derived transferred T cells, and failed to show any difference between the two cell types. Because of these conflicting data, we have reexamed the persistence of adoptively transferred viral antigen specific T-cells derived from TCM and TEM population. Accordingly, we developed a NOG mouse model for studying the ability of human CMVpp65-specific T cells derived from central memory and effector memory populations to migrate to and accumulate in human tumor xenografts expressing CMVpp65, to alter the growth of these tumors and to persist in the tumors. This model also allows us to test immunomodulating agents and their ability to enhance targeted T-cell accumulations, antitumor activity and persistence. We analyzed CMVpp65-specific CD8 T cells derived from TCM and TEM precursors in vitro and in vivo. To tract the T-cells in vivo, we transduced membrane-bound Gaussia luciferase into TCM and TEM populations and monitored T cell trafficking by in vivo bioluminescence. Contrary to expectation, our results initially showed no differences between TCM and TEM derived CMVpp65-specific T-cell in mice co-treated with IL-2 in the time to accumulation, ultimate level of accumulation, degree of CMVpp65+ tumor regression or T-cell persistence. However, in mice cotreated with IL-15/IL-15Rα complex, both TCM and TEM exhibited more sustained engraftment and more prolonged accumulation in both the targeted tumor and in the marrow. In mice treated with IL-15/IL-15Rα, TCM and TEM derived T cells showed a similar effector memory phenotype and a similar level of regression of tumor growth. Thus, adoptive transfer of CMVpp65 specific TCM or TEM when combined with IL-15/IL-15Rα complex may support better persistence of antigen-specific T-cells following adoptive immunotherapy. Studies comparing IL-15/IL-15Rα complex with IL-15 alone are in progress.

Disclosures

No relevant conflicts of interest to declare.
Spontaneous Apoptosis of the Lymphocytes in Cord Blood of Late Preterm Newborns
Authors: Wasiluk Alicja , Zak Janusz , Warda Janusz
Published: October 2019

Background/Aims: Apoptosis plays an important role in shaping the repertoire of lymphocytes and in regulating the size of the mature lymphocyte pool. The best-defined regulators of apoptosis in T cells are members of the Fas and Bcl families. Fas/FasL pathway is critically involved in the elimination of mature, self-reactive lymphocytes. Bcl-2 protein is important for lymphocyte survival in response to growth factors and various activating stimuli. We have compared the role of CD4+, CD8+, Fas, FasL and Bcl-2 in spontaneous apoptosis of T lymphocytes from cord blood and tested there is or not relation to gender.

Patients and Methods: We have introduced 28 late preterm newborns to our studies, 15 females and 13 males. Blood was collected from umbilical artery immediately after cutting cord. We have determined the following parameters: the per cent of CD4+ and CD8+ lymphocytes, the Fas and FasL phenotype and Bcl-2 using flow cytometer Coulter PC 500, Krefeld, Germany.

Results: We have found the increase per cent of CD4+ in females 56.10 %, in relation to males 38.15 %. The same per cent was found for CD8+ in both gender: 19.60 % females, 17.35 % males.

The antigen Fas (CD95) was more expressed in females: on CD4+ 12.80 %, CD8+ 8.15 %, in comparison to males: CD4+ 9.89 %, CD8+ 6.06 %. FasL (CD95L) was lower per cent in females: both on CD4+ 6.92 %, CD8+ 5.68 %, in relation to males: CD4+ 8.94 %, CD8+ 7.04 %. Bcl-2 was a little lower in females on CD4+ 96.9 % and evidently lower on CD8+ 79.14%. In males we noted on CD4+ 97.40% and on CD8+ 93.01 %. Summarizing our findings we can underline T lymphocytes in females are more activated, what indicate Fas. Inhibitor of apoptosis Bcl-2 is less expressed in females, in comparison to males lymphocytes.

Conclusion: Our preliminary results seems to indicate spontaneous apoptosis of T lymphocytes is more expressed in female newborns.

Table I. Spontaneous apoptosis of the T lymphocytes in cord blood of late preterm newborns. N Gender Lymphocyte T % Fas % FasL % Bcl-2 % 15 Female CD4+ 56.10 12.80 6.92 96.90 13 Male 38.15 9.89 8.94 97.40 15 Female CD8+ 19.60 8.15 5.68 79.14 13 Male 17.35 6.06 7.04 93.01

Disclosures

No relevant conflicts of interest to declare.
The "Platelet Boilermaker" Is an Option for Quickly Raising Platelet Counts in IVIG Refractory ITP
Authors: Deloughery Thomas G.
Published: October 2019

Most patients with immune thrombocytopenia (ITP) respond to first line therapy. Rarely there are patients who remain with very low platelet counts despite administration of intravenous immune globulin (IVIG). A protocol at our institution (colloquially known as the "platelet boilermaker") for these refractory patients is to give 1 gram/kg of IVIG continuously over 24 hours concurrent with one unit of plateletpheresis product given over 6 hours repeated times 4. We examined the charts of 10 patients in the past 6 years who have received combined therapy. 70% were male; all were refractory to steroids and IVIG. Results are in table: Abstract 5023. TableAgeGenderDiseaseBleedingPlts PreDay +1Day+2Day+3Results65MEvansGIB1110510Stopped bleeding59MITP, CLLBruising3656No response34FITPPregnancy2583175150Delivered81FITPEpistaxis15384167Splenectomy day +123FITPEpistaxis552165Splenectomy day +164MDITPEpistaxis, ruising252427Stopped bleeding89MITPEpistaxis4655046Splenectomy day +157MITPBruising9586946Splenectomy day +128MITPICH14133325326No extension78MITPGIB27354498Splenectomy day +1

Plt = platelet count x 109/L

DITP = drug induced ITP, ICH - intracranial hemorrhage. GIB - gastrointestinal bleeding

9/10 patients had at least a transient rise in platelets greater than 30x109/L - 5 patients underwent emergent splenectomy with no bleeding complications. The patient with ICH had no extension and while the other patients who responded had cessation of bleeding. In summary for patients with IVIG refractory ITP who either have bleeding or need urgent splenectomy combined continuous platelets/IVIG is a therapeutic option.

Disclosures

Off Label Use: profilnine - warfarin reversal rVIIa - warfarin reversal.
Analysis of Using Low Dose 3 Factor PCC (Bebulin®) for INR Reversal in Patients with Warfarin Induced Severe/Life-Threatening Bleeding
Authors: Mekan Sabeen F. , Logdberg Lennart , Stein Arline , Nash Leah , Cohen Jessica , Rosenberg David
Published: October 2019

Background: 3 Factor Prothrombin Complex Concentrates have been used off label with plasma products for effective and timely reversal of INR in patients on Warfarin presenting with severe bleeding. The exact dosage required for this purpose has yet to be determined.

Methods: We conducted a retrospective review of all patients taking Warfarin who presented to our institution between January 1, 2013 to June 30, 2014 who received 3 Factor Prothrombin Complex Concentrate (PCC) Bebulin® for INR reversal for significant/lifethreatening bleeding. The dose of Bebulin was determined by clinical judgment by the blood bank and the treating physician in accordance with the patient’s weight, bleeding location and the presenting INR.

Results: A total of 116 patients were treated. Mean age was 72 years. Forty-two (36.21%) of patients were female. Most common bleeding site was intracranial (n=61), followed by systemic hemorrhage (n=24), GI bleeding (n=16), trauma (n=10) and other bleed (n=5). Some patients were also on other anticoagulants besides Warfarin – 28 patients (24%) were on Aspirin and 9 (7.8%) on Plavix. The mean INR prior to Bebulin treatment was 3.62 which dropped to 2.05 at an average of 2.2 hours post-treatment. Mean level at 12 hour post-Bebulin dropped to 1.57. The average patient received 2 units of plasma and 1 unit of packed red cell transfusions. The average Bebulin dose administered was 959 IU (SD +/-465 IU) with average patient weight of 82.9 kg (SD +/- 27kg). Based on this the average Bebulin dose administered was 11.57 IU/kg. Eighty-eight patients (76%) were able to be discharged home or to a skilled nursing facility.

Thirty-six patients (31.1%) required more than one dose of Bebulin®. Mean INR at presentation in this group of patients were not similar to the remainder of the group. Mean INR at average of 2.5 hours post-Bebulin in this group was 2.18 which dropped further to an average of 1.75 at 9 hours. The total average Bebulin dose incrementally administered was similar in both groups.

Conclusion: Low dose Bebulin at doses of approximately 12 IU/kg with 2 Units of plasma was effective in timely reversal of INR in majority of patients who presented with Warfarin induced bleeding.

Disclosures

Off Label Use: Bebulin (3 Factor PCC) is approved for use in Hemophilia patients; we discuss using low dose for INR reversal in Warfarin associated bleeding.
Down-Regulation of HO-1 Promoted Apoptosis Induced By Decitabine Via Increasing p15INK4B Promoter Demethylation in myelodysplastic syndrome
Authors: Wang Jishi , Ma Dan , Fang Qin , Wang Ping , Gao Rui , Sun Jia
Published: October 2019

Background: Decitabine, reverts hypermethylation of p15INK4B gene in vitro, was used to improves cytopenias and blast excess in over 50% of patients with high-risk myelodys plastic syndrome (MDS). In this study, over-expression of Heme Oxygenase-1(HO-1) was found in MDS cells line SKM-1 cells, and it was closely related to resistance to apoptosis induced by decitabine.

Objective: we aimed to further investigated what role of HO-1 exactly played in apoptosis induced by low-does of decitabine in MDS.

Method: CCK-8 kits was used to determine the proliferation inhibition of SKM-1 cells. Flow cytometry was used for analyzing cell proliferation rate and apoptosis. The methylation status and expression of P15INK4B in mRNA and protein levels were measured by methylation-specifc polymerase chain reaction (PCR [MSP]). Apoptosis relative factors expression were detected by real-time transcription and Western blot.

Result: Up-regulation of HO-1 by transfected it into SKM-1 cells via lenti-virus vector promoted proliferation and protected cells against apoptosis. In contrast, down-regulation of HO-1 enhanced decitabine-induced apoptosis but reduced accumulation of S phase in cell cycle. To explore the mechanism, we detected cell cycle relative protein expression after SKM-1 cells were treated by decitabine in each group. As a result, over-expression of p15 INK4B and CDK4 were observed in SKM-1 cells which HO-1 was inhibited. And p15 INK4B and CDK4 expression-dependent apoptosis was related to caspase3 pathway. Even though HO-1 was silenced, but apoptotic rate never increased as caspase3 pathway was blocked.

Conclusion: As we known that p15 INK4B is a keypoint to regulate S phase of cell cycle, in our study, more obvious demethylation of p15 INK4B was seen in group of SKM-1 cells in which HO-1 was down-regulated. It’s equally in patients’ mononuclear cells who suffered from MDS. The worse the prognosis of MDS was judged, the more the mRNA level of HO-1 expressed. In conclusion, over-expression of HO-1 indicated resistance to demethylation of p15 INK4B induced by decitabine.

Disclosures

No relevant conflicts of interest to declare.
Wogonin Inhibits Angiogenesis Via a c-Myc/Hif-1α–dependent Pathway in Multiple Myeloma
Authors: Fu Rong , Guo Qinglong , Chen Yan , Song Huihui , Chen Baoan
Published: October 2019

Wogonin, a plant-derived flavone, has been shown to inhibit tumor angiogenesis. Angiogenesis is associated with multiple myeloma (MM) progression. However, the mechanisms that Wogonin inhibits angiogenesis in MM microenvironment are not well known. In this study, we investigated Wogonin inhibited tumor angiogenesis that correlated with the inhibition the secretion of vascular endothelial growth factor (VEGF) in MM cells. The c-Myc/Hif-1α–dependent pathway was essential to regulate MM cell-mediated angiogenesis and contributed to tumor growth. Wogonin inhibited c-Myc activation and down-regulated the expression of c-Myc and Hif-1α. Functionally, based on transient knockdowns and overexpression, our data delineate Wogonin inhibited VEGF production and secretion mediating tumor angiogenesis via c-Myc/Hif-1α–dependent pathway. Our data, therefore, identify Wogonin as a novel anti-angiogenesis compound to inhibit tumor promotion in MM.

Disclosures

No relevant conflicts of interest to declare.
Recovery Blasts in the Peripheral Blood in AML Patients Who Obtain a Complete Remission Is a Favorable Prognostic Indicator
Authors: Cook Nathaniel M. , Butera James N. , Reagan John L
Published: October 2019

Introduction: Many patients with Acute Myelogenous Leukemia (AML) will transiently develop blasts in the peripheral blood upon recovery of induction chemotherapy, despite going on to obtain a complete remission. These are presumably non-leukemic marrow blasts. Recovery blasts can sometimes reach over 20% in the peripheral blood in remission patients. The significance of recovery blasts cells after induction chemotherapy in patients with AML is not clear. We sought to define the incidence of recovery blastocytosis (RB) and determine if the presence of RB has prognostic relevance.

Methods: We performed a retrospective analysis of 72 patients with newly diagnosed AML who underwent induction chemotherapy with 7+3 (cytarabine and daunorubicin) with or without etoposide between the years of 2006 and 2013 at our institution. Patients were included in the analysis as having RB only if their blasts resolved by day 14 of induction chemotherapy, blasts reappeared upon recovery after day 14, recovery blasts were > 5% in the peripheral blood of a duration of >5 days, and subsequently resolved by day 45 and remission was obtained. We recorded patient's absolute and percentage peripheral blast counts on presentation, upon nadir and throughout recovery of induction chemotherapy. The incidence and level of RB in patients who achieved a remission was tabulated. This group (group 1) was compared to the patients who achieved a remission whom did not develop RB (group 2).

Results: 45/77 AML patients obtained a CR after induction chemotherapy. Recovery blasts were seen in 9/45 (20%) remission patients (group 1), with a median RB of 9% for a median duration of 8 days. Of these 9 patients, 4 developed RB > 10%, and 2 developed RB >20 % Group 1 did not differ in respect to age, gender, cytogenetic profile, etoposide use, or whether the patient received 1 or 2 induction therapies from the 36/45 remission AML patients who did not develop RB (group 2). Median survival for group 1 was = 1,376 days, and 1,036 days for group 2 (p= .035).

Conclusion: Recovery Blasts in the peripheral blood after induction chemotherapy is seen commonly in AML patients who obtain a complete remission. RB can be seen even as high as > 20% in remission patients. AML patients who develop RB on their way to remission after induction chemotherapy have a better overall survival then patients who do not develop RB.

Figure 1 Figure 1.

Graph 1. Kaplan-Meier survival curves for AML patients with RB (group 1) and without RB (group 2).

Disclosures

No relevant conflicts of interest to declare.
TGFβ and IL10 Have an Impact on Risk Group and Prognosis in Childhood ALL
Authors: Wiegering Verena , Taschik Julia , Haubitz Imme , Eyrich Matthias , Winkler Beate , Schlegel Paul Gerhardt
Published: October 2019

Introduction: Considering that cytokines play an important role in immune response and that many infectious, autoimmune and malignant diseases are influenced by cytokine production, we hypothesized that genetically determined cytokine gene polymorphism might have an important influence on prognosis in pediatrics acute lymphoblastic leukemia (ALL).

Methods: In this study, 95 pediatric ALL-patients were examined with regard to cytokine gene polymorphisms (TNFα, TGFβ, IL10 and IFNγ) and their potential association with prognosis. Moreover we analyzed the intracellular production of theses cytokines in patient T-cells.

Results: TGFβ high-producer-haplotypes were associated with high-risk ALL-patients (Codon 10: T/T) and with the tendency of a reduced overall survival, whereas IL10 high-producer-genotypes were associated with a reduced relapse rate and a superior overall survival compared to IL-10-low-producer patients. Gene-polymorphisms of the pro-inflammatory cytokines IFNγ and TNFα did not show an impact on prognosis and risk-group of ALL in our cohort. On a functional basis TNFα and IFNγ expression of T-cells at initial diagnosis was significantly reduced in high-risk- and T-ALL-patients in comparison to healthy controls.

Summary: Cytokine gene-polymorphisms of the regulatory/anti-inflammatory cytokines TGFβ and IL10, but not of the pro-inflammatory cytokines IFNγ and TNFα seem to have an impact on prognosis of pediatric ALL patients. Reduced capacity to produce pro-inflammatory cytokines at diagnosis may serve as a functional risk factor. These data may help in further risk stratification and adaptation of therapy-intensity.

Disclosures

No relevant conflicts of interest to declare.
The Prevalent, Features and Prognostic Impact of Deletion p16 in Patients with Adult Acute Lymphoblastic Leukemia
Authors: Xu Na , Li Yulin , Zhou Xuan , Xiaoli Liu , Zhou Hongsheng , Huang Fen , Xu Bing , Liu Qifa , Sun Jing , Feng Ru , Meng Fanyi
Published: October 2019

Background and objective: Deletion of chromosome 9p21 is a crucial event for acute lymphoblastic leukemia (ALL). 9p21.3 genes encode three cell cycle inhibitory proteins: p15INK4b, p16CDKN2A, and p14ARFT. Recently studies show that p16 (CDKN2A) alleles were hypermethylated at CpG islands in ALL patients and delation p16 was associated with poor prognosis in childhood ALL. However, the prognostic significance of the deletion p16 in adult ALL leukemia is controversial, so the aim of the present study was to investigate the prevalence, feature and specific prognostic relevance of delation p16 in Chinese ALL patients from a single center in China.

Patients and Methods: A total of 513 newly diagnosed adult ALL patients were identified retrospectively from the database of ALL between January 2008 and December 2013 at our center. We Detected delation p16 by interphase fluorescence in ituhybridization (I-FISH) and analyzed their clinical data retrospectively.

Results: Of 513 cases, the prevalence of having either heterozygous or homozygous p16 deletions was 32%. p16 deletion were identified in 27% of newly diagnosed Philadelphia positive(Ph+) patients by univariate analysis, patients with p16 deletion had no significant difference compared with wild-type patients in terms of sex, age, white blood cells (WBC) count at diagnosis,BM blast percentage,chromosome karyotype,extra infiltration and CR I rate. The patients with p16 deletion was more likely to relapse comparaed with wild-type patients (P=0.03), and at relapse, we found a strong trend in the detection rate of p16 loss (41%) compared with diagnosis (P=0.01), suggesting that loss of this genomic region may be involved in disease progression. Of note, in newly diagnosed Ph+ALL with delation p16 were likely relapse even in the patients who treated with allogeneic hematopoietic stem cell transplantation (P=0.03). In addition, deletion p16 were significantly associated with poor outcomes in terms of overall survival (P=0.01), disease free-survival (P< 0.001), and cumulative incidence of relapse (P< 0.001).

Conclusion: In our study, the absence of p16 expression seems to a poor prognostic marker in adult ALL patients. However, how to improve the survival of these patients need further study.

Disclosures

No relevant conflicts of interest to declare.
Modeling the Epidemiology of Indolent Non-Hodgkin Lymphoma (iNHL) in France
Authors: Troussard Xavier , Herrera Alain , Cornet Edouard , Solal-Céligny Philippe
Published: October 2019

As of 2014, there were few reliable publication sources of epidemiology data for Non-Hodgkin Lymphoma (NHL) in France, particularly with regard to indolent forms such as Follicular Lymphoma (FL), Waldenström Macroglobulinemia (WM) and Marginal Zone Lymphoma (MZL). In addition, epidemiology registries provide useful data on disease incidence but no information can be directly obtained on the number of relapsed patients, especially in the rare iNHL subtypes. At the same time, many new and effective drugs will be made available to the Hematologist/Oncologist community. As a consequence, there will be an increase in the complexity of therapeutic algorithms.

We undertook an innovative modeling approach to address the need for a greater understanding of the number of relapsed patients available to these new therapies. The objective of the model was to determine the yearly incidence of first line and later lines of therapy patients from 2014 to 2018 and in consequence, provide relevant data to determine precisely which population might be in need for the new compounds and bring accurate data that could be implemented into a prospective economic model.

First, we used the Registre Régional des Hémopathies Malignes de Basse-Normandie (RRHMBN) to collect real world patient characteristics, such as patient subgroups and tumor staging. The RRHMBN is dedicated to Hematological malignancies and supported by official Health Authorities: Institut National du Cancer(INCA) and Institut National de Veille Sanitaire (INVS). Then, we applied these data to the French National Network of Registers FRANCIM incidence and survival data (table 1 below).

Table 1 Yearly number of patients newly diagnosed Table 1. Yearly number of patients newly diagnosed

Finally, we applied the results from medical literature to progression free survival for each standard of care treatment in our epidemiological model.

The table below shows yearly the number of patients in need for treatment in first and subsequent lines of therapy as produced by our model.

Table 2 Yearly number of patients in need for treatment in first and subsequent lines Table 2. Yearly number of patients in need for treatment in first and subsequent lines

Our modeling approach compensates for a lack of real world information on relapsed iNHL. We have shown that there is a significant number of relapsed patients who are in need of new treatments in France. The model is able to answer the question of how many patients are in need of novel therapies in later lines through 2018 and provide for iNHL investigators efficient prospective data that can be integrated into medico-economic model development.

Furthermore, these data address a current medical need for the development of a new iNHL algorithm, necessitated by the expected arrival of new treatment options for patients in the relapsed setting.

Disclosures

No relevant conflicts of interest to declare.
Bendamustine-Rituximab (BR) Combination in Low Grade B-Cell Lymphoproliferative Disorders (LPD): A Community Oncology Experience
Authors: Shahab Aarish , Chin Janet , Yunus Saadia
Published: October 2019

Introduction:

Low-grade B-Cell LPDs are indolent Non-Hodgkin’s Lymphoma (NHL) that are incurable with current therapeutic options. BR chemoimmunotherapy has demonstrated remarkable effectiveness in these patients. Here we report our experience with BR in low-grade B Cell LPD subtypes.

Method:

Between October 2011 and May 2014, we treated 25 patients with a diagnosis of low-grade B-Cell LPD with a planned BR regimen of 6 cycles. The median age was 64 years (range 41-91), 15/10 M/F – a ratio of 3:2. LPD subtype included Chronic Lymphocytic Leukemia (CLL) 5, Small Lymphocytic Lymphoma (SLL) 3, Follicular Lymphoma (FL) 6, Marginal Zone Lymphoma (MZL) 2, Mantle Cell Lymphoma (MCL) 2, Mucosa-associated Lymphoid Tissue (MALT) 3, and Lymphoplasmacytic Lymphoma (LPL) 4. Twenty two (88%) were stages III or IV. Ten had received prior treatments. 15 had BR as first treatment. The regimen consisted of Bendamustine (B) 90 mg/m2 (on days 1&2 and Rituximab (R) 375 mg/m2 on day1 given every 28 days.

Results:

Seventeen (68%) completed the entire 6 planned cycles. 14 (56%) required treatment delays due to various toxicities. In 5 (20%) chemo was discontinued early for toxicity. The most common non-hematologic toxicity was fatigue 11 (44%), and skin rash 5 (20%). The grade III/IV hematologic toxicity was as follows neutropenia (3) anemia (9) and thrombocytopenia (2).

The overall response rate (ORR) was 92% (23 pts) with 18 (72%) Partial Responses (PR), and 5 (20%) Complete Responses (CR). Twenty four (96%) continued to have remission with no progression noted during follow-up. One patient with 17p del CLL progressed after 3 months with Richters transformation to diffuse large B cell NHL. The median duration of response was 17 months, with a range from 2 months – 38 months.

Conclusion:

In all LPD subtypes, despite the observed toxicities requiring treatment delays and dose modifications, 96% patients attained significantly long lasting remissions. Although this review included a comparatively small number of patients, our experience shows that BR is a highly effective chemo immunotherapy in all subtypes of Low-Grade B-Cell LPD – suggestive of altering the natural biology of this disease. For more definitive results, the study should be inclusive of a larger number of patients and prolonged follow-ups.

Disclosures

No relevant conflicts of interest to declare.
Vemurafenib As First Treatment of Hairy Cell Leukemia
Authors: Cheng Gregory , Cheng Gym , Chan Natalie Pui Ha , Wong Chris , Wong Raymond S
Published: October 2019

44 years old lady presented with fever and pancytopenia in March 2014.CXR showed bilateral pneumonia. Hb 7.5xg/dl, plt 42x x109/l, wbc 1.2 x19neutrophils22%, lymphocytes75%.Circulating lymphocytes with hairy cytoplasmicprojections and indented nuclei were noted. Flow cytometry showed these abnormal lymphoid cells were CD19+ve, CD5-ve, CD 23-ve, FMC7+ve, CD25+ve, CD11c+ve and CD103+ve.Bone marrow biopsy revealed a hypercellular marrow with dense infiltration of lymphoid cells of the same immunophenotype. Braf V600 mutation was detected. Cladribine or pentostatin was out of stock and import of these drugs would take at least 2 months. In view of the severe pancytopenia and on -going infection, various treatment options were discussed with, the patient .Patient decided to start on vemurafenib 960mg twice daily while awaiting cladribine. After 8weeks of treatment, the peripheral blood counts were normalized. Hb 12.2g/dl, plt 153x x109/l, wbc 3.1x109/l, neutrophils 53%,lymphocytes 40%. Braf V600 mutation was no longer detected. Vemurafenib was well tolerated and the patient received treatment mainly as outpatient. Vemurafenib was discontinued after 8 weeks and the patient then received a 5-day course of cladribine. She remained in complete remission

Discussion

Vemurafenib had shown to be safe and effected in hairy cell leukemia patients who were refractory to or who relapsed after purine analogs.1,2 Still to be determined are the correct vemurafenib dosing strategy, the best timing , duration and scheduling of vemurafenib. Due to unusual circumstances, our patient received 8 weeks of vemurafenib as first line therapy. The patient achieved a complete remission with minimal residual disease. Follow data is needed to see how long the patient remains in remission

1. N Engl J Med 2014; 370:286-288

2. 19th Congress of the European Hematology Association (EHA): Abstract S696.

Disclosures

Off Label Use: Vemurafenib as first treatment of Hairy Cell Leukemia. Wong:johnson &johnson: Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding; GlaxoSmithKline: Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding, Speakers Bureau; Biogen-Idec: Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding; Novartis: Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding, Speakers Bureau; Bayer: Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding; Pfizer, MSD, Roche, BMS, Baxter, Amgen, Alexion: Research Funding.
Pre-Clinical Evaluation of a Novel DNA Crosslinking Agent, Bo-1055 in B-Cell Lymphoma
Authors: Caldas Lopes Eloisi , Correa Fabian , Peguero Elizabeth , Ambati Srikanth R. , Shieh Jae Hung , SU Tsann Long , Moore Malcolm A.S.
Published: October 2019

Some B-cell lymphoma including mantle cell lymphoma (MCL) and diffuse large B-cell lymphoma (DLBCL) remain incurable using conventional chemotherapeutic approaches. Therefore it is important that new treatment strategies be developed. We have evaluated the efficacy of a number of novel, third generation, DNA-directed alkylating agents that have DNA specific binding domain by linking DNA-affinic molecules to N-mustard pharmacophore via a urea, carbamate or hydrazinecarboxamide linker (Kapuriya et al. Bioorganic & Medicinal Chem. 19:471–485; 2011). One of these, the water-soluble ureidomustine BO-1055, was screened for toxicity against a panel of human lymphoma cell lines including MCL (JEKO-1, Z-138, HBL-2), DLBCL (OCILy10, OCILy19) and a spontaneous murine B-cell lymphoma. We also screened BO1055 against a panel of normal human cells including mesenchymal stromal cells (IC50 >10uM), bone marrow-derived endothelium (IC50 >10uM) lung basal epithelium, bronchial epithelium and myofibroblasts (IC50s >10uM) and purified cord blood CD34+ cells in suspension culture or colony-forming assay (for hematopoietic progenitor cells) (IC50 >10uM) and in cobblestone area-forming assay (for hematopoietic stem cells) (IC50 9.1uM). The mean IC50±SD (uM) in MCL cell lines was JEKO-1 (0.266±0.27), Z-138 (0.182±0.15), HBL2 (0.161±0.34), In DLBCL lines OCILy10 (0.117±0.21), OCILy19 (0.287±0.17) and murine B-cell lymphoma (0.463±0.39). Our results indicated that BO-1055 has a significant therapeutic window (50-100-fold) between its toxicity against human B-cell lymphomas compared to various normal human cell types. We evaluated BO-1055 cardiotoxicity in the HL-1 cardiomyocyte line and observed a 227-fold less cytotoxicity compared to Doxorubicin. Treatment with BO-1055 resulted in accumulation of cells in S-phase and up-regulation of proteins involved in DNA repair [MRE11, p-P95/NBS1 (ser343), RAD50, p-ATR (ser428)] while Bcl-6, an important B-cell lymphoma biomarker, was down-regulated. Xenograft experiments in NSG mice bearing JEKO-1 GFP/luciferase+ tumors treated with BO-1055 (30mg/kg) 3x/week showed complete tumor remission after 2 weeks of treatment as monitored by luciferase image. Our results suggest that BO-1055 has potent activity against B-cell lymphoma and present a strong rationale for its further therapeutic development as an alkylating agent due to his low toxicity against normal tissue and high toxicity against hematopoietic tumors.

Disclosures

No relevant conflicts of interest to declare.
the -137G/C Polymorphism in Interleukin-18 Gene Promoter Contributes to Chronic Myeloid Leukemia Risk in Turkish Patients
Authors: Yalcin Serap , Mutlu Pelin , Cetin Turker , Sarper Meral , Ozgur Gokhan , Avcu Ferit
Published: October 2019

Interleukin-18 (IL-18) is a cytokine which belongs to the interleukin-1 superfamily and secreted by various immune and non-immune cells. Evidence has shown that IL-18 has both anticancer and procancer effects. The aim of this study was to evaluate the relationship between -137G/C (rs187238) IL-18 gene polymorphism and susceptibility to chronic myelogenous leukemia(CML) in Turkish patients. The frequencies of polymorphism -137G/C were studied in thirty chronic myelogenous and thirty healthy individuals. The genotyping was performed by polymerase chain reaction and DNA sequencing analysis. Significant associations were detected between the IL-18 rs187238 (G/C) polymorphism and CML patients. A higher prevalence of the C allele was found in CML cases with respect to controls (p<0.05). The GC heterozygote and CC homozygote genotypes are associated with the risk of CML when compared with controls (OR, 1.9023; 95% CI, 0.6171-5.8636 for GC genotype and OR, 10.3585; 95% CI, 0.5326-201.4622 for CC genotype). These results indicate that individuals that are either heterozygote (GC) or homozygote for CC genotype are associated with the development of chronic myelogenous leukemia in Turkish population. Further large-scale studies combined with haplotype and expression analysis are required to validate the current findings.

Disclosures

No relevant conflicts of interest to declare.
Spectrum of BCR-ABL Kinase Domain Mutations; A Cohort Study from Saudi Arabia
Authors: Abalkhail Halah , Khalil Salem
Published: October 2019

Chronic myeloid leukemia (CML) is a clonal myeloproliferative neoplasm (MPN) associated with a characteristic translocation between chromosome 9 and 22 to form Philadelphia chromosome (Ph). The introduction of tyrosine kinase inhibitors (TKIs) imatinib for CML marked a new era for targeted therapy. In general CML patients in chronic phase (CP) have a very good response to Imatinib with a ~85% projected overall rate of survival. However, a significant proportion harbor molecular residual disease and develop acquired resistance after initial response through a number of cellular and molecular mechanisms in which mutations in the Abl kinase domain are the best known cause of resistance. Mutations are detected in 35% of patients with resistance to TKI in Chronic Phase and up to 80% in patients with accelerated phase and blast phase.

Given the clinical importance of the mutation status, mutation analysis was established as a clinical service to aid in the clinical decision and identify the subset who would benefit from alternative therapies.

In this retrospective study, we assessed the ABL kinase domain mutation in 93 patients referred from our clinics at King Faisal Specialist Hospital and Research centre (General Organization) between 2011-2013 with Ph positive CML displaying either refractory to TKI or resistance displaying increase BCR-ABL1 levels by serial monitoring using Quantitative PCR. Mutation analysis was performed on RNA extracted from either blood or Bone marrows after amplification of the BCR/ABL1 transcript by nested PCR followed by Direct sequencing of the BCR-ABL1 Kinase domain including residues 243-487.

In total, 93 patients (46 Females and 47 males), Age between 10-71 years (median age: 38 years) of which 89 were adults and 4 pediatrics were analyzed. Among 93 patients, 20 (21.5%) were positive for 10 different mutations across the ABL1 Kinase domain mutations (7 patients had T315I, 3 patients had each of the following mutation: Y253H, F359I, 2 patients with E255K. Additionally, one patient with each of the following mutations: E355G, V299L, L248V, L298, Y326H and F317L). The duration from diagnosis to mutation detection ranged between 3-144 months with a median duration of 4 years.

Despite the retrospective nature and the relatively small sample size of a single center analysis, the mutation frequency is in line with similar studies reported from our region.

Further follow up is needed to identify the outcome of these patients acquiring ABL kinase mutations on targeted therapy.

Disclosures

No relevant conflicts of interest to declare.
Pharmacoeconomical Analysis of Chronic Myelogenous Leukemia Treatment Free Remission
Authors: Shuvaev Vasily , Abdulkadyrov Kudrat , Turkina Anna , Martynkevich Irina , Chelysheva Ekaterina , Fominykh Mikhail , Martchenko Valentin
Published: October 2019

Introduction. Implementation of target therapy in chronic myelogenous leukemia (CML) treatment allowed saving life for majority of CML patients. First and second generation tyrosine kinase inhibitors (TKI) usage experience showed that part of patients can achieve deep molecular responses. Clinical trials of treatment cessation in CML patients with deep molecular response demonstrated that substantial proportion of patients can be free from drug with predictable relapse risk and safe treatment re-initiation. This approach is interesting due to sustained increasing number of CML patients and avalanche like increment of budget burden because of continuous expensive TKI therapy.

The aim of our study was to assess potential budget impact of successful TKI cessation in CML patients with long-lasting deep molecular response.

Materials and methods. Pharmacoeconomical modelling of CML diagnostic and treatment process including treatment free remission (TFR). Markov chain approach was implemented to construct models for non-stop therapy and TFR in patients with sustained molecular response 4 logs (MR4.0) (Fig 1). Treatment re-initiation was considered in case of major molecular response loss. The total number of CML patients in Russia was 6500; annual rate of newly diagnosed CML cases was 800 (in accordance with Russian CML Registry data). The patient consent rate to TFR inclusion was assumed as 80%. Transition rates were chosen from clinical trials and own experience. Cost-utility analysis of first and second generation TKI apply was performed. Simulation of clinical and economical consequences was done on national level. Five-year (2015-2020) time horizon was used. To simplify presentation of our results we recalculated costs to US dollars.

Results. Our analysis showed that by year 2020 the estimated total CML patient number in Russia will be about 11000 patients with total direct yearly CML cost 109 mln $. TFR approach can decrease budget burden from 109 to 95 mln $ yearly and the total evaluated money saving during years 2015-2020 period in Russia will be 67 mln $ with complement cost-utility data (Fig 2).

Conclusion. Annual CML cost will increase due to increment of CML patients number. Nevertheless, TFR strategy in CML patients with long-lasting deep molecular response under continuous molecular monitoring can significantly decrease the healthcare budget burden. TFR approach has the advantage over sustained therapy on financial cost and efficacy of its use.

Disclosures

Turkina: Bristol Myers Squibb: Consultancy; Pfizer: Consultancy; Novartis Pharma: Consultancy.