bluuebunny/crossref_metadata_2025_split huggingface dataset:
Characterization Of An Acquired IgG Autoantibody To Bβ and γ Chains Of Fibrinogen Resulting In Delayed Fibrin Polymerization and Severe Bleeding
Authors: Lulla Premal , Dasgupta Swapan , Rice Lawrence , McCarthy John J. , Thiagarajan Perumal 
Published: October 2019

Abstract
A 88 year-old woman was referred to our center for 3 weeks of gastrointestinal bleeding and a spontaneous right rectus femoris hematoma. The prothrombin time (PT), activated partial thromboplastin time (aPTT), thrombin time (TT) and reptilase time (RT) were markedly prolonged (Table 1). Assays for coagulation factors V, VIII, IX and X were normal. A fibrinogen concentration of < 50 mg/dl was measured by Clauss method while fibrinogen concentration measured by radial immunodiffusion was 180 mg/dl. Following infusions of cryoprecipitate there was accelerated clearance of fibrinogen as measured by the Clauss method. An inhibitor of fibrinogen function was suspected.Table 1TestTime (sec)1:1 mix (sec)Normal Range (sec)PT>902212-15aPTT70.73923-36TT>120>12015-19Reptilase Time>120>12014.8-20.4
When the patient’s plasma was tested towards immobilized fibrinogen in ELISA plates, a concentration dependent specific binding of IgG was seen while under similar conditions control IgG had no significant binding (Figure 1). The IgG antibody was polyclonal comprising of both kappa and lambda subtypes.
The IgG fraction was purified on a Protein A column and the isolated IgG fraction prolonged the thrombin time of normal plasma in a concentration dependent manner. The effect of the isolated IgG fraction on fibrin polymerization was measured by monitoring the absorbance at λ390 in a spectrophotometer. The IgG inhibited fibrin polymerization in a concentration dependent manner. Control IgGs had no significant effect on polymerization under these conditions (Figure 2).
The immunologic specificity of antibody was tested in immunoblots of fibrinogen subjected to SDS-PAGE. The patients IgG reacted with fibrinogen in unreduced buffers. Under reducing conditions, the IgG reacted with Bβ and γ chains of fibrinogen while the control IgG did not (Figure 3).
The patient was treated with rituximab and steroids. After four, weekly doses of rituximab, fibrinogen levels by Clauss method normalized (450 mg/dl). There was no detectable antibody to fibrinogen and no further bleeding was noted.

Conclusion
This is a description of a rare case of a polyclonal autoantibody to Bβ and γ chains of fibrinogen that inhibited fibrin polymerization leading to a severe bleeding diathesis.

Disclosures:
No relevant conflicts of interest to declare.
Thrombomodulin Alfa Treatment For Disseminated Intravascular Coagulation In Acute Promyelocytic Leukemia: A Retrospective Analysis Of The Open-Label, Multicenter Study Cohort
Authors: Matsushita Tadashi , Watanabe Jyunichi , Honda Goichi , Mimuro Jun , Takahashi Hoyu , Tsuji Hajime , Eguchi Yutaka , Kitajima Isao , Sakata Yoichi 
Published: October 2019

Abstract

Background
Patients with acute promyelocytic leukemia (APL) patients present disseminated intravascular coagulation (DIC) that can result in life-threatening hemorrhagic complications. In 2008, the anticoagulant thrombomodulin alfa (TM-α, recombinant human soluble thrombomodulin) was approved for the treatment of DIC in Japan. After commercial launch, only small case series have been described in APL patients, and data from larger patients cohort are awaited. We accessed the clinical safety and beneficial effects of TM-α treatment in DIC patients with APL, by evaluating data on the open-label, multicenter, post-marketing surveillance study cohort.
Patients and Methods
A retrospective evaluation was carried out on a cohort of 172 patients with APL from the open-label, multicenter, post-marketing surveillance study of TM-α in Japan. Of all 172 patients, 31 were relapse/refractory APL patients and 141 were newly diagnosed. The newly diagnosed APL patients were generally received all-trans retinoic acid (ATRA) and anthracycline-containing induction therapy followed by consolidation therapy as previously reported by the Japan Adult Leukemia Study Group (Yanada et al, Eur. J. Haematol. 2007). The relapse/refractory APL patients were received arsenic trioxide, tamibarotene, and/or gemtuzumab ozogamicin. Early death (death within 30 days from the start of antileukemic treatment), severe hemorrhagic events, and the improvement of coagulopathy were analyzed.

Results
The study population consisted of 97 males and 75 females, with a median age of 58 (range 10 - 88). TM-α was administration by 380 ± 52.5 U/kg/day and the duration of dosing was 7.0 ± 6.3 days. In most patients (81 %), TM-α was started on or before the start of antileukemic treatment. Within the first 30 days, 12 patients (7 %) had severe hemorrhage and there were 24 (14 %) early deaths. Six of those (3.5% of 172 patients) were due to hemorrhage. TM-α treatment rapidly and significantly improved coagulopathy regardless of concomitant ATRA treatment (Table 1).

Conclusions
Although anticoagulation therapy during APL chemotherapy may induce severe hemorrhagic complication, our findings suggest that supportive care with TM-α ameliorates coagulopathy and reduces the risk of hemorrhagic early deaths in patients with APL.

Disclosures:
Watanabe: Asahi Kasei Pharma Corporation: Employment. Honda:Asahi Kasei Pharma Corporation: Employment.
Characterization Of Chromatin Dynamics In Mammalian Erythropoiesis
Authors: Zhao Baobing , Keerthivasan Ganesan , Yang Jing , Ji Peng 
Published: October 2019

Abstract
Mammalian erythropoiesis is a dynamic process in which erythroid progenitors proliferate and differentiate into mature enucleated red blood cells. In the late stage of erythropoiesis, erythroblasts undergo terminal cell-cycle exit, chromatin condensation and extrusion of the pycnotic nucleus via an asymmetric cell division. Recent genetic and biochemical studies illustrated that various signaling pathways, including histone deacetylations and other chromatin modifications, are involved in chromatin condensation and enucleation. However, it is unclear the global dynamic changes of the nucleosome and how different histones are regulated during chromatin condensation.
We proposed to directly characterize the expression levels and localization of different histones and their variants during erythropoiesis. We found that histone H2A and its variants are differentially expressed during erythropoieisis, while the levels of other histones including H2B, histone H3 and histone H4 remained unchanged. To determine their localization, we performed immunofluorescence experiments of these histones together with nuclear lamins. Strikingly, lamin B staining revealed a gradually enlarged nuclear opening through which most histones, except histone H2AZ, were released out of the nucleus. This nuclear opening was confirmed by electron microscopy. The same phenotype was observed in vivo in freshly purified late stage erythroblasts. Western blot analysis confirmed this release, however, many well known nuclear proteins remained in the nucleus in this process. Mechanistically, we demonstrated that caspase 3 was involved in the regulation of nuclear openings during erythropoieiss. Inhibition or knockdown of caspase 3 completely blocked lamin B opening and histone release, which led to inhibition of chromatin condensation and cell differentiation, and ultimate cell death.
In summary, our study demonstrated that the dynamic changes of various histones, with their selective and gradual release out of the nucleus, is required for chromatin condensation and differentiation during erythropoieisis. Studies of the detailed mechanisms are ongoing. Particularly, we are using micrococcal nuclease digestion of chromatin followed by next generation sequencing technology (MNase-seq) to determine genome-wide nucleosome location during the differention. Chromatin- immunoprecipitation (ChIP) will also be performed to determine H2A family protein locations during erythropoiesis.

Disclosures:
No relevant conflicts of interest to declare.
Deletion Of Samsn1 Underlies Genetic Susceptibility To Monoclonal Gammopathy Of Undetermined Significance (MGUS) In Mice
Authors: Amend Sarah , Chu Liang , Serie Daniel , Vachon Celine M , Lu Lan , Vij Ravi , Colditz Graham A , Weilbaecher Katherine N , Tomasson Michael H. 
Published: October 2019

Abstract
KaLwRij/C57Bl (KaLwRij, “Kal-ridge”) is a spontaneous mutant mouse strain predisposed to monoclonal gammopathy of uncertain significance (MGUS) and parallels many features of human pathology including progression to multiple myeloma (MM). In African Americans, an increased prevalence of MM is due to an increased risk of MGUS development (rather than increased rate of progression from MGUS) and is likely therefore mediated by germline MGUS risk alleles. In both humans and mice, the genetic basis for MGUS/MM risk is unknown. We sought to identify the genetic basis for MGUS/MM risk in KaLwRij mice to shed light on genes or pathways that might mediate risk in humans. We employed a novel integrative computational analysis, combining KaLwRij/B6 modified haplotype block analysis and low-power human MM/control single nucleotide polymorphism (SNP) data. We used high-density SNP arrays to establish genetic distances of KaLwRij from 11 another diverse inbred strains and confirmed close homology to parent C57Bl/6. Haplotype block analysis identified genetic loci that diverged from C57Bl/6 and included over 500 genes. We filtered the haplotype list using previously published RNA expression data from the bone marrow of C57Bl/6 and KaLwRij mice, to identify 71 differentially expressed genes. As a final filter, we used SNP data from a genome wide association study (GWAS) of human Caucasian MM case and normal volunteer samples (∼300 each) to identify SNPs differentially represented in MM patients. Combining these sets yielded four (4) candidate genes. Our top candidate, Samsn1 (Hacs1), is a negative regulator of B-cell activation and interestingly, was first identified as a gene differentially expressed in MM. Strikingly, we found that KaLwRij mice had a complete germline deletion of the Samsn1 gene. Samsn1 is expressed within the hematopoetic compartment, including B-cells, macrophages, and osteoclasts (OC), key cell types that participate in MM pathology. Consistent with the reported inhibitory function of SAMSN1, KaLwRij had enhanced naïve B-cell response and constitutive Lyn phosphorylation. Further, we found that KaLwRij macrophages and OC also have constitutive Lyn activation consistent activation of cytokine signaling pathways in stromal cells. Absence of Samsn1 may lead to unregulated B-cell and macrophage/OC activation, priming both a pro-tumor bone microenvironment and B-cells for development of MGUS and predisposition to MM. Our approach focused on germline loci rather than malignant cell genetic alterations, advancing our understanding of host predisposition. Identification of the gene or genes predisposing patients to MGUS/MM will provide novel targets for identifying at-risk individuals and will potentially allow the develop of preventative therapeutics for MM.

Disclosures:
No relevant conflicts of interest to declare.
Vitamin D Effect On Umbilical Cord Blood Characteristics: A Comparison Between African Americans and Caucasians
Authors: Eldjerou Lamis K , Rosenau Emma , Lu Xiaomin , Bennett Clayton , Sugrue Michele W. , Hoyne Jonathan , Lambert Amy , Hertel Lindsay , Sazama Kathleen , Wingard John R , Zubair Abba C 
Published: October 2019

Abstract

Background
Umbilical cord blood (UCB) is an important alternative source of stem cells for patients who lack matched adult donors, particularly in ethnic minorities. The infused total nucleated cell (TNC) and CD34+ cell dose are important prognostic factors on transplant outcomes. UCB units collected from African Americans (AA) have lower TNC and CD34+ cell counts and more likely to be disqualified for banking compared to other racial and ethnic groups. Furthermore, AA, including pregnant women, have increased prevalence of vitamin D deficiency. Vitamin D has an established role in hematopoiesis; therefore, we investigated the racial differences in UCB vitamin D content and its correlation with UCB cell composition and hematopoietic potential.

Methods
119 UCB units that did not meet the TNC count banking criteria were analyzed. 51 UCB units were collected from AA mothers and 68 from Caucasian mothers. We analyzed UCB variables including volume, hematocrit (HCT), TNCs, mononucleated cells (MNC), CD34+ cells, plasma 25-Hydroxy vitamin D [25(OH)D] and in vitro progenitor cell hematopoietic potential measured by Colony-Forming Cell (CFC) assay.

Results
The median values of 25(OH)D in UCB units were significantly lower in AA compare to Caucasians (p= <.0001). Severe vitamin D deficiency < 10 ng/mL was exclusively detected in UCB units from AA (19/51; 37%) and plasma 25(OH)D concentration < 20 ng/mL was more prevalent in AA (43/51 units; 84%), compare to Caucasians (15/68 units; 22%). UCB units from AA had lower absolute numbers of TNC (p= .0016), MNC (p= .0261), and CD34+ cells (p= .0257) compare to Caucasians. No difference in median CFC/10,000 MNC was detected between AA and Caucasians (p= 0.661) (Table 1).
Univariate analysis of 25(OH)D effect on UCB variables revealed a correlation between 25(OH)D level and TNC (r= 0.193, 95% CI 0.013-0.36; p= .0353) and UCB HCT (r= 0.196, 95% CI 0.016-0.363; p= .0327). A significantly higher MNC count was noted when 25(OH)D concentration was ≥ 20ng/mL (4.5x 108, range (1.7-9.5) vs. 3.9x 108, range (0.8-9); p= .0329). This correlation between 25(OH)D level and TNC and MNC was not detected after adjustment for race (Figure 1). However, 25(OH)D concentration ≥ 20 ng/mL significantly correlated with CBU HCT in AA race (median 31.9% (24-42) vs. 27.8%, range (21.4 - 40); p= .0124).

Conclusion
25(OH)D level, TNC and HCT are all significantly lower in UCB units from AA compare to Caucasians. Independent of race, 25(OH)D directly correlated with TNC and MNC. These data suggest vitamin D and other yet to be determined factors play an important role in the mechanism of low UCB cell and progenitor counts.

Disclosures:
No relevant conflicts of interest to declare.
Carfilzomib, Rituximab and Dexamethasone (CaRD) Is Highly Active and Offers a Neuropathy Sparing Approach For Proteasome-Inhibitor Based Therapy In Waldenstrom’s Macroglobulinemia
Authors: Peter Treon Steven , Tripsas Christina K , Meid Kirsten , Cropper Steven J , Mostyn Patrick , Warren Diane , Paba-Prada Claudia E. , Patterson Christopher , Yang Guang , Hunter Zachary , Kunsman Janet , Kanan Sandra , Ghobrial Irene M. 
Published: October 2019

Abstract
Bortezomib is active in Waldenstrom’s macroglobulinemia (WM) but associated with considerable peripheral neuropathy (PN). The proteasome inhibitor carfilzomib (CFZ) is approved in the USA for relapsed/refractory myeloma. Herein, we examined the efficacy and safety of carfilzomib, rituximab, and dexamethasone (CaRD) in 31 proteasome inhibitor and rituximab naive WM patients with symptomatic disease. Median baseline characteristics: age 61, prior therapies 0 (range 0-1), hematocrit 32.3%, hemoglobin 10.7 g/dL, serum IgM 3375 mg/dL, serum M-protein 2.185 g/dL, B2M 3.6 mg/L, and bone marrow disease involvement 60%. Therapy consisted of IV CFZ 20 mg/m2 (cycle 1) then 36 mg/m2 (cycles 2 and beyond) with IV dexamethasone (dex) 20 mg given on days 1,2,8,9 and rituximab 375 mg/m2 on days 2,9 of each 21-day cycle. Treatment consisted of six induction cycles, then maintenance beginning 8 weeks after induction (given every 8 weeks for eight cycles; consisted of CFZ 36 mg/m2, and IV dex 20 mg on days 1,2 and rituximab 375 mg/m2 on day 2). Patients with IgM level >4000 mg/dL underwent plasmapheresis and/or had rituximab held until IgM <4000 mg/dL to prevent symptomatic IgM flare. Patients received oral acyclovir (400 mg twice daily) and famotidine (20 mg twice daily) as concomitant medications. For all 31 patients, median serum IgM levels and M-protein declined to 749 mg/dL and 0.7 g/dL, respectively (p<0.00001). Median hematocrit and hemoglobin rose to 40.9% and 13.7 g/dL, respectively (p<0.00001). A total of 30 patients concluded induction therapy with bone marrow tumor involvement reduced to a median of 7.5% (p=0.0003). The best overall response rate using criteria adapted from the 3rd International Workshop on WM was 81% (1 CR, 8 VGPR, 12 PR, 4 Minor Responses). With a median follow-up of 8 cycles, 22 patients remain on study, including 20 currently on maintenance therapy. Median time to response (for minor responders or better) was 2.1 months. Grade >2 treatment related toxicities included asymptomatic elevation in lipase (12.9%), dex-related hyperglycemia (6.45%), reversible neutropenia (9.67%), and cardiomyopathy (3.22%). There were no grade 2 or greater PN events. Treatment discontinuation occurred for non-response (n=8), cardiomyopathy in a patient with multiple cardiac risk factors (n=1), and progressive disease (n=1). CaRD is highly active and offers a neuropathy sparing approach for proteasome-inhibitor based therapy in WM.

Disclosures:
Off Label Use: Carfilzomib is a novel proteasome inhibitor, which offers a neuropathic sparing approach on waldenstrom macroglobulinemia disease when used in a combination therapy with rituximab and dexamethasome.
Whole Genome and Exome Sequencing Defines The Genetic Landscape Of Hepatosplenic T-Cell Lymphoma
Authors: Scotland Paula , Gaulard Philippe , Love Cassandra L , Fataccioli Virginie , Travert Marion , De Leval Laurence , Weisenburger Dennis D. , Czader Magdalena , Parihar Mayur , Nair Reena , Sengar Manju , Beaven Anne W , Crow Jennifer H. , Miles Rodney R. , Gordon Leo I. , Chadburn Amy , Evens Andrew M , Gill Javed , Fedoriw Yuri D. , Richards Kristy L. , Srivastava Gopesh , Choi William W. L. , Flowers Christopher R. , Bernal-Mizrachi Leon , Mann Karen P. , Naresh Kikkeri , Hsi Eric D. , Horna Pedro , Tao Jianguo , Sun Zhen , Long Kevin , Zhang Jenny , Dave Sandeep 
Published: October 2019

Abstract

Background
Hepatosplenic T-cell lymphoma (HSTL) is a rare form of lymphoma, comprising less than 1% of the cases. However, HSTL extracts a highly disproportionate toll on patients with a median age of diagnosis of 35 years and an expected median survival of less than two years. The vast majority of HSTL patients eventually succumb to their disease. The genetic basis of the disease is largely unknown. Although abnormalities of chromosome 7, including isochromosome 7q occur commonly in the disease, the role of specific genes and genetic mutations to the disease remains essentially unknown.

Methods
In this study, we sought to define the genetic features of HSTL through the whole genome sequencing and exome sequencing of 32 HSTL tumors and germline DNA (where available) from the same patients. Exome enrichment of DNA was carried out using the Agilent solution-based system of exon capture, which uses RNA baits to target all protein coding genes as well as ∼700 human microRNAs. Both whole genome and exome sequencing were performed using the Illumina platform.

Results
We identified 28 candidate cancer genes that were recurrently mutated in HSTL. Commonly implicated biological processes comprising these genes included signal transduction (e.g. PIK3CD, KRAS) and chromatin modification (e.g. TET1, SETD2 and MLL3), accounting for 16% and 23% of the total genetic events, respectively. Nearly all of these genes have been implicated in HSTL for the first time and provide new insights into the pathogenesis of the disease and potential targets for therapy. Whole genome sequencing confirmed isochromosome 7q as the most common recurrent chromosomal abnormality in HSTL and additional structural genetic alterations in chromosome 7.

Conclusion
Our study provides the most comprehensive genetic portrait of HSTL to date, and is a significant step in defining the genetic causes of this disease.

Disclosures:
No relevant conflicts of interest to declare.
Next-Generation Sequencing Suggests Complex, Heterogeneous Pathogenesis In Peripheral T-Cell Lymphoma Unspecified
Authors: Schatz Jonathan H. , Horwitz Steven M. , Lunning Matthew A. , Dolgalev Igor , Huberman Kety , Heguy Adriana , Viale Agnes , Socci Nicholas D. , Pirun Mono , Teruya-Feldstein Julie , Wendel Hans-Guido 
Published: October 2019

Abstract
Peripheral T-cell lymphoma (PTCL) makes up about 12 percent of non-Hodgkin lymphoma, comprising 18 diseases that are poorly understood and carry a generally worse prognosis than B-lymphomas. PTCL not otherwise specified (PTCL-NOS), a diagnosis of exclusion, is most common, making up 25-30 percent. Gene-expression studies suggest a heterogeneous origin of this diagnosis, with overlap to other PTCL types, but the genetic factors underlying its pathogenesis are undefined. Current therapy for PTCL-NOS is empiric and ultimately ineffective for most patients. Identification of specific therapeutic targets is therefore a high priority. We have sought better understanding of pathogenesis through next-generation sequencing of PTCL-NOS tumor DNA. Whole-exome sequencing revealed candidate genes but low availability of fresh-frozen samples limited our ability to draw conclusions by this method alone. We therefore sequenced the coding regions of 237 candidate genes in a collection formalin-fixed paraffin-embedded samples. We used Nimblegen Sequence Capture for PCR amplification of exons and Illumina hiSeq for raw sequence generation. Results were aligned to hg19 and compared to dbSNP and the 1,000 genomes data to exclude germline variants. Analysis, including comparison to the COSMIC database of cancer-specific mutations, revealed high-confidence mutations affecting more than 60 known cancer-related genes in 25 PTCL-NOS cases. Recurrent mutations pointed to frequent activation of three key signaling pathways: NF-kB (TNFAIP3), WNT/B-Catenin (APC, CHD8, CELSR2), and NOTCH (NOTCH1, FBXW7). Recurrent deregulation of epigenetic processes was indicated by mutations in genes affecting histone acetylation (EP300, CREBBP), histone methylation (MLL2, KDM6A), and DNA methylation (TET2, DNMT3A). In addition, components of core tumor suppressor pathways showed evidence of frequent inactivation (TP53, ATM, RB1, CUL9, PRKDC). In all, 22 of 25 cases had mutations in at least one of these 17 recurrently mutated genes. Multiple additional candidate disease mechanisms also were suggested by lower-confidence mutations but require confirmation studies, which are under way. In sum, analysis of the coding region of PTCL-NOS tumor DNA suggests a complex and heterogeneous pathogenesis, in line with gene-expression profiling. This work provides an opportunity to better sub-classify entities within the diagnosis of PTCL-NOS and identify specific therapeutic targets and their associated biomarkers.

Disclosures:
Horwitz: Seattle Genetics, Inc.: Consultancy, Research Funding; Millennium: Consultancy, Research Funding.