bluuebunny/crossref_metadata_2025_split huggingface dataset:
MUTATIONAL STATUS of REARRANGED IMMUNOGLOBULIN HEAVY CHAIN VARIABLE Genes in a COHORT of SLOVENIAN PATIENTS with CHRONIC LYMPHOCYTIC LEUKAEMIA.
Authors: Pajič Tadej , Maleŝiĉ Ivan , Zver Samo , Ĉernelĉ Peter 
Published: October 2019

Abstract
Abstract 4399

Introduction
The aims of the study were to determine the mutational status and IGHV gene usage in our cohort of patients with B-CLL and compare the results to those published in the literature.

Patients and Methods
57 B-CLL patients (22 female, 35 male), median age 65 (range from 37 to 88 years) in various Binet stages (47 Binet A, 6 Binet B, 4 Binet C) were included in the study. All the patients were untreated at the time of blood collection. Leukaemia cells were isolated from venous blood samples of patients by Ficoll density gradient centrifugation. Total cellular RNA was isolated from leukaemia cells and used to the prepare cDNA by reverse transcription. After heteroduplex analysis, clonal PCR products were identified with polyacrylamide gel electrophoresis. The PCR products were sequenced and mutational status was determined by comparing the sequence of the IGHV region of the patient sample to the most homologous germ line V sequence. Sequence that was homologous more than 98 % with their corresponding germ line sequence was considered unmutated.

Results
Unmutated and mutated IGHV mutational status in our cohort of patients with B-CLL was found in 42 % (N=24) and 58 % (N=33), respectively.
At the IGH subgroup level, the most frequently used subgroup was IGHV3 (38,55 %), followed by IGHV1 and IGHV4, found in 28,10 % and 22,85 %, respectively. Subgroups IGHV2 and IGHV5 were found out in 2 patients each (3,5 %). The other subgroups, IGHV6 and IGHV2, were determined in one patient each. At the specific gene level, the most frequently used gene IGHV was IGHV1-69, found out in 15,85 %. Genes V4-34, V4-39 in V3-23 were determined in 7,05 %, followed by genes V3-07, V3-11, V3-15, V3-30 and V3-48 identified in 5,25 % each. The others were identified at the lower frequency.
In patients with unmutated IGHV genes, the most frequently observed gene was V1-69 (33,3 %). The most frequently used gene in mutated rearrangements was IGHV4-34 (12,13 %). The gene V3-21 was not found out in our cohort of patients.

Conclusions
In the study performed, we have found out that the portion of unmutated IGHV genes of the clonal B lymphocytes, the gene usage and somatic mutations frequency were similar to those published in the literature. Interestingly, the gene V3-21 was not found out in our cohort of patients, the results that is similar to the Mediterranean-based study. In the future, the geographic comparison and more definitive conclusions could be made on the population based investigations of IGHV gene use in large series of CLL patients in Slovenian region.

Disclosures:
No relevant conflicts of interest to declare.
Resveratrol Plus Simvastatin Are Effective in Decreasing IgM Secretion in Asymptomatic Waldenstrom Macroglobulinemia. One Year Follow-up.
Authors: Iuliano Francesco , Infusino Stefania , Perricelli Alessia , Di Maio Massimo , Pomillo Angelo , Luci Maria , Mingrone Francesco 
Published: October 2019

Abstract
Abstract 4401

Background
Waldenström macroglobulinemia (WM) is a distinct B-cell lymphoproliferative disorder characterized by lymphoplasmacytic bone marrow infiltration along with an immunoglobulin M (IgM) monoclonal gammopathy. Asymptomatic patients with monoclonal IgM and without morphologic evidence of bone marrow infiltration < 10% clonal marrow cells) are classified as having IgM-MGUS.Therapy is postponed for asymptomatic patients, and progressive anemia is the most common indication for initiation of treatment. Resveratrol (3,4',5-tri-hydroxy-trans-stilbene) is an antioxidant constituent of a wide variety of plant species including grapes. It has gained considerable attention because of its anticancer properties, as shown in solid and hematologic malignancies. Published data show that resveratrol has significant antitumor activity in WM cells line. Moreover, simvastatin, a 3-hydroxy-3-methyl-glutaryl-CoA reductase inhibitor, induced inhibition of proliferation, cytotoxic effect and apoptosis in IgM secreting cell lines as well as in primary CD19(+) WM cells. With this background we have treated 4 patients with asymptomatic WM with an association of simvastatin and resveratrol to test the efficacy of such of drugs in asymptomatic Waldenstrom macroglobulinemia

Methods
4 pts (3 males and 1 female), median age 42,3 yrs (range, 42–73) and asymptomatic WM were treated with a schedule containing resveratrol 40 mg/die and simvastatin 20 mg/die for at least 90 days. At enrollment patients characteristic were hemoglobin level median, 12.1 g/dL,serum beta(2)-microglobulin level median, 2.4 mg/L, and IgM peaks median, 1.8 g/dL. All patients have taken regularly the drugs and there have been no adverse events.CK and LDH serum levels were kept in the normal range.

Results
In all IgM-MGUS patients a reduction of more than 50% of the IgM peak was observed after 3 months of therapy and it was still maintained at 12 months of follow-up. In SWM patient the reduction was about 25% and it was manteined over time. Striking, another patient with Waldentrom disease resistant to the previous therapy with EDX and anti-CD20 MoAb achieved a CR only after adding resveratrol and simvastatin.

Conclusions
Our data demonstrate clearly that the association between resveratrol with simvastatin decreases IgM secretion in Waldenstrom macroglobulinaemia and can be useful in asymptomatic or low risk patients not having any adverse effects.

Disclosures:
Off Label Use: Simvastatin showed in vitro activity on waldentrom cell lines Resveratrol showed in vitro activity on waldenstro cell lines.
KIR Genes and Their HLA-C Ligands in B-Cell Chronic Lymphocytic Leukemia in a Polish Population.
Authors: Frydecka Irena , Jedynak Anna , Karabon Lidia , Pawlak Edyta , Tomkiewicz Anna , Kielbinski Marek , Woszczyk Dariusz , Wolowiec Dariusz , Kuliczkowski Kazimierz 
Published: October 2019

Abstract
Abstract 4402

Introduction
Dysfunction in cellular and humoral immunity entails an increased risk of B-cell chronic lymphocytic leukemia (B-CLL). It has been suggested that innate immunity, especially natural killers cells play key role in antitumor cytotoxicity regulated by interaction between killer immunoglobulin-like receptors (KIRs) on NK cells and some T-cell subpopulations and their HLA-class I ligands on target cells. KIRs receptors have been divided into two functional groups: with long cytoplasmic tails (DL) - inhibitory and with short cytoplasmic tails (DS) - activating. Two broad haplotypes of KIR genes have been defined: the A haplotype characterized by presence of single activating KIR gene (2DS4) and the B haloptype characterized by two or more activating KIR genes (2DS1, 2DS2, 2DS3, 2DS5 and 3DS1).The 2DL2, 2DL3 and 2DS2 KIRs bind to HLA-C1 allotype (Asp80), while 2DL1 and 2DS1 KIRs bind to HLA-C2 allotype (Lys80). Due to lack of representative data in literature the study was undertaken to determine the association between polymorphism of KIR genes and HLA-C allotypes and susceptibility to B-CLL.

Patients and methods
The following KIR genes were typed: 2DL1, 2DL2, 2DL3, 3DL1, 2DS1, 2DS2, 2DS3, 2DS4fl, 2DS4del, 2DS5, 3DS1, in 200 B-CLL patients and 199 healthy individuals using PCR-SSP method. The HLA-C1 and HLA-C2 were determined using Olerup SSP typing kit in 187 B-CLL patients and 104 controls.

Results
Individual KIR gene content was similar in B-CLL cases and healthy controls although patients with KIR2DS3 gene were more common among cases compared to controls (36% vs. 27%, p=0.06) (Table1). The frequency of AA haplotype did not differ between studied groups (29% vs. 28%). Moreover no differences were found between incidence of HLA-C allotypes (HLA-C1: 74.3% vs. 76.0%, HLA-C2: 67.4% vs.70.2%), genotypes (C1/C1: 32.6% vs 29.8%, C1/C2: 41.7% vs 46,2% and C2/C2: 25.7% vs 24.1) and KIR genes in combination with genes for their HLA-C ligands in B-CLL patients and controls (Table 2).

Conclusions
The data obtain in this study imply that there may be not direct association between KIR and HLA-C gene content in the genome and the presence of B-CLL.

Disclosures:
No relevant conflicts of interest to declare.
Acadesine (Acadra‘) for Patients with Resistant/Relapsed Chronic B-Cell Lymphocytic Leukemia (B-CLL): A Multicentre Phase I/II Study.
Authors: Van Den Neste Eric W , Gonzalez-Barca Eva , Janssens Ann , Cazin Bruno , de Oteyza J. Pérez , Terol Maria Jose , Auton T. , Saunders Andrew , Campas Clara 
Published: October 2019

Abstract
Abstract 4405
Acadesine (Acadra‘) is a nucleoside that induces cell death in B-cell chronic lymphocytic leukemia (B-CLL) cells ex vivo in a dose-dependent manner over the concentration range 50 mM to 1 mM. Acadesine enters B-cells and is phosphorylated to its active metabolite, ZMP, which induces apoptosis independently of ATM or p53. It is active against B-CLL cells from patients who have not responded to prior treatment with fludarabine and/or chlorambucil (Campas et al, 2003).
Based on this pre-clinical data, a Phase I/II clinical study was designed to determine the safety and tolerability of acadesine given intravenously as a 4-hour infusion to patients with B-CLL. This is an open-label dose escalation study of acadesine with two parts. In Part I, the dose escalation part of the study, patients receive a single dose of acadesine on Day 1 and are followed up to Day 22. In Part II, patients will receive up to 5 doses of acadesine at the maximum tolerated dose (MTD) identified in Part I over a period of up to 20 days starting on Day 1. The patient population includes B-CLL patients with refractory or relapsed disease who have received one or more (≥ 1) prior lines of treatment which must have included either a fludarabine or an alkylator based regimen. Primary endpoints of the study evaluate the safety and tolerability of acadesine. Secondary endpoints evaluate the pharmacokinetics of acadesine in plasma, its metabolite ZMP in whole blood, and changes in peripheral blood B-cell and T-cell counts as efficacy biomarkers. Optimal Biological Dose (OBD) is defined as the minimal acadesine dose that gives a maximum Cmax for ZMP in whole blood with a good safety profile.
Twelve patients (6 males and 6 females) have been administered acadesine to date in 3 cohorts in Part I. Acadesine has an excellent safety profile to date. The principal safety finding was asymptomatic acadesine-related hyperuricaemia of short duration in four patients. Prophylactic allopurinol has significantly reduced the incidence of hyperuricaemia.
Pharmacokinetic data showed acadesine rapidly disappears from plasma to be converted into ZMP inside blood cells. The OBD has not been reached yet and maximum dose administered to date is 139.5 mg/kg. The next cohort will receive 210 mg/kg. Additional safety, pharmacokinetics and efficacy data will be presented at the meeting.

Disclosures:
Off Label Use: Acadesine (AcadraTM): nucleoside that induces cell death in B-cell chronic lymphocytic leukemia.. Auton:BTG Limited: Consultancy; Advancell Technologies Inc.: Consultancy. Saunders:BTG limited: Consultancy; Advancell Technologies: Consultancy. Campas:Advancecell Technologies Inc: Employment, Equity Ownership.
Thromboembolic Complications of Intravenous Immunoglobulin (IVIG) in Hypogammaglobulinemic Patients with Chronic Lymphocytic Leukemia (CLL).
Authors: Milani Cannon , Dalia Samir , Colvin Gerald A. 
Published: October 2019

Abstract
Abstract 4416
Infectious complications represent a major cause of morbidity and mortality in patients with chronic lymphocytic leukemia (CLL). The etiology is postulated to be secondary to aberrations in cell-mediated immunity, as well as to therapy-related immunosuppression.
Hypogammaglobulinemia, which occurs in virtually all patients with CLL, may beprofound and correlates with disease duration and stage. In 1981 during treatment of two children with hypogammaglobulinemia and coincidental idiopathic thrombocytopenic purpura (ITP), physicians in Switzerland observed a reproducible increase in the platelet count following IVIG treatment. This clinical observation and follow up systemic investigations further intensified the widespread clinical use of IVIG as a potential immune modulatory agent. Intravenous immunoglobulin (IVIG) therapy has been used successfully to prevent and treat infections in this cohort of patients.
Today, Intravenous immunoglobulin (IVIG) is used in a broad spectrum of autoimmune, inflammatory, and primary and secondary immunodeficiencies. The efficacy has been demonstrated in several control studies. IVIG is a blood product prepared from the serum of between 1,000 and 15,000 donors per batch. The mechanism of action validated by in-vitro models is exerted by a combined effect on autoantibodies, complement activation, cytokines, and saturation of Fc receptors on tissue macrophages.
However IVIG administration and treatment is not benign and should be used with caution given the potential manifestations of thromboembolic complications. High concentration and rapid infusion rate of the IVIG, as well as increased dose and osmolarity of the solution are thought to predispose to thrombotic events. Serum viscosity is the implicated mechanism for compromised blood flow and predisposition of high-risk patients to cardiovascular or cerebrovascular infarction.
In this retrospective review, we detail IVIG related thromboembolic manifestations in CLL patients, to highlight the importance of risk stratifying patients prior to treatment administration. The current consensus surrounding IVIG is that of a relatively safe treatment, with minor adverse effects such as hypertension, fever and chills, nausea, myalgias, or headache.
However our report highlights the importance of proceeding with caution in the application of this therapy, as it's proclivity for thrombotic complications has not been fully elucidated in patients with underlying malignancies. Pre-existing thrombogenic risk factors should be carefully evaluated in patients undergoing treatment with IVIG. Clinical evaluation, with careful attention to vascular history and underlying co-morbidities can potentially unmask the high-risk patient where IVIG could be lethal.

Disclosures:
No relevant conflicts of interest to declare.
Dysregulation of Metabolic Genes May Be Normalized by Epigenetic Drugs (Vorinostat, Decitabine) and Nutritional Compounds (Genistein, Vitamin D) in Leukemic Cells.
Authors: Karlic Heidrun , Varga Franz , Pittermann Elisabeth , Pfeilstocker Michael 
Published: October 2019

Abstract
Abstract 4423

Background
Dysregulation of energy metabolism is a key feature of malignancy and known to be regulated by epigenetic mechanisms. This study explores gene expression in the KG1 cell line and a primary culture of leukemic blasts from a patient, both incubated with SAHA (suberoyl anilide hydroxamic acid, also known as Vorinostat) in comparison to other epigenetic drugs such as Decitabine (DAC) but also nutritional compounds such as genistein and vitamin D3 with a known activity on metabolism where epigenetic mechanisms play a role.

Methods
Following a 3 days incubation with pharmacologic concentrations of the above mentioned compounds, gene expression patterns of the KG1 cell-line were analysed on an Affymetrix Gene Expression Array and specifically evaluated by quantitative real time PCR from KG1 cells and primary leukemic cells of a patient with an AML-M2 which were cultured at the same conditions as the KG1 cells. In order to confirm epigenetic mechanisms induced by targeted drugs, methylation patterns of selected tumor suppressor genes were analyzed using specific primers for bisulfite-treated DNA.

Results
Our experiments show that expression of a large scale of micro RNAs which are known to be responsible for alternative splicing is stimulated by SAHA as well as DAC and to a lesser extent also Vitamin D3 and genistein, but downregulation of mitochondrial RNAs as a consequence of apoptosis was exclusively observable by SAHA.
HDAC-inhibition results in clear repression of metabolic genes involved in glycolysis (5/6 genes) as well as cholesterol-biosynthesis (14/15 genes) and fatty acid synthesis (all 4 genes) which are also known to be regulated by nutrition. Classic tumour suppressor genes, transcription factors and cell-cycle regulators such as P53, P15, P16 and P21 and FOXO were (re-) activated with this treatment while MYC was down-regulated.

Discussion
A downregulation and thus normalization of a large scale of genes including key genes from malignancy-associated stimulation of glycolysis as well as biosynthesis of cholesterol and fatty acids was observed with SAHA and nutritional compounds such as Vitamin D and genistein. We propose that these interactions should be carefully considered in clinical application, and a combination of synergistic drugs and/or dietary strategies may provide a major advantage in future drug development.

Acknowledgment
Supported by Jubilaeumsfonds der Oesterreichischen Nationalbank.

Disclosures:
No relevant conflicts of interest to declare.
Results of a World Wide Survey On the Diagnostic Dilemma Between Type 2B Von Willeband Disease and the PT-VWD.
Authors: Othman Maha , Hamilton Alexander , Lillicrap David 
Published: October 2019

Abstract
Abstract 4436
The Canadian PT-VWD project (www.pt-vwd.org) initiated in 2007 is based on a molecular diagnostic approach to differentiate between type 2B VWD and its closely similar disorder PT-VWD. Recruitment of worldwide samples and data still poses a challenge partly because of the relative rarity of the two bleeding disorders and more importantly because it is dependant on the level of awareness about the requirements for the differential diagnosis and the subsequent impact on the treatment decision. The objective of this survey was to gather an opinion from the worldwide hemostasis community about this diagnostic dilemma and the current utilization of the appropriate diagnostic tests. The survey comprised eight key questions (see link on website) sent individually by email to hematologists from different countries and representing a variety of academic centres. Out of 60 emails we received 37 responses. The highest response was from Europe 16/37 followed by USA 9/37. Other responses were from Africa (3), Canada (2), Asia (2), Middle East (2), South America (1), Mexico (1) and Australia (1). Fifty four percent of specialists have seen or diagnosed 1-5 type 2B VWD cases and 38% have seen or diagnosed 1-5 PT-VWD cases. 82% have reported they have access to laboratory phenotypic tests including RIPA and/or platelet mixing studies with only 59% reporting they have access to genetic testing of the two responsible genes: VWF and platelet GP1BA gene. Genetic analysis was the most discriminative test in the opinion of 54% and the RIPA was chosen to be the discriminative test by 35% while 11% believed the appropriate discrimination should be based on both. 84% agreed to classify PT-VWD under platelet function disorders and 73% have felt that PT-VWD should be investigated when platelet function defects are suspected. On a scale from 1-10 with 10 being most important, 30/37 of participants reported 7-10 to rate the importance of this diagnostic dilemma and 13% have reported 5. Despite the relatively small number of respondents, the data obtained through this survey confirm the rarity of the two disorders, reflects the positive world-wide opinion re the diagnostic challenges of the two closely similar disorders and indicate the significant unavailability of genetic analysis as a useful confirmatory test.

Disclosures:
No relevant conflicts of interest to declare.
Therapeutic Algorithm for Patients with Acquired Von Willebrand Syndrome and Monoclonal Gammopathy.
Authors: Staritz Peter , Krause Manuela , Zimmermann Rainer , Huth-Kuehne Angela 
Published: October 2019

Abstract
Abstract 4445
In contrast to congenital von Willebrand Syndrome therapy of acute bleeding in acquired von Willebrand Syndrome (AWS) with von Willebrand factor containing concentrates (F VIII/vWF) is often ineffective. In patients with IgG monoclonal antibodies therapy with intravenous immunoglobulins (IVIG) is reported to be effective whereas patients with IgM antibodies seem not to respond to IVIG.
Over the last years we diagnosed and treated several patients with AWS either prior to surgery or due to acute bleeding. About half of the patients responded adequately to IVIG as reported in the literature. One patient with no response to IVIG 4 days after the last dose received DDAVP with a significant increase in von Willebrand factor-antigen and ristocetin cofactor and normalized half life. Dental surgery could be performed without any bleeding complications under daily infusions of DDAVP. Another patient with partial response to IVIG and contraindications to DDAVP received F VIII/vWF prior to dental extraction. In contrast to former recoveries with a shortened half life of the infused concentrate (about 2 hours), half life of vWF was significantly prolonged following prior IVIG treatment. Another patient responded sufficiently to single therapy with F VIII/vWF. A further patient with suspected diagnosis of moderate hemophilia A had received DDAVP for severe epistaxis with good response and sufficient half lifes. He was transferred to our centre and we diagnosed AVW with IgM antibodies. We performed another recovery and half life with DDAVP with adequate response.
According to our experience we propose the following therapeutic algorithm: A recovery with DDAVP should be performed first line if treatment is not contraindicated. In case of insufficient half lifes and/or intended major surgery, a recovery and half life with F VIII/vWF concentrate should follow. In case of significantly reduced half life of wWF an attempt with IVIG is necessary. If response to IVIG is not adequate further treatment with DDAVP or F VIII/vWF is indicated.
As treatment with IVIG is not predictable in all patients with IgG antibodies and is very costly a therapeutic attempt with DDAVP in the first line and F VIII/vWF second line is worthwhile. In major surgery requiring prolonged replacement therapy, IVIG treatment, if effective, is less costly than treatment with FVIII/vWF.

Disclosures:
No relevant conflicts of interest to declare.