bluuebunny/crossref_metadata_2025_split huggingface dataset:
Array Comparative Genomic Hybridization of Ribosomal Protein Genes In Diamond-Blackfan Anemia Patients; Evidence for Three New DBA Genes, RPS8, RPS14 and RPL15, with Large Deletion or Duplication
Authors: Gazda Hanna , Landowski Michael , Buros Christopher , Vlachos Adrianna , Sieff Colin A. , Newburger Peter E. , Niewiadomska Edyta , Matysiak Michal , Glader Bertil , Dobson Lori , Atsidaftos Eva , Lipton Jeffrey Michael M. , Beggs Alan 
Published: October 2019

Abstract
Abstract 1007
Diamond-Blackfan anemia (DBA) is an inherited bone marrow failure syndrome characterized by anemia, usually presenting during infancy or in early childhood. Although anemia is the most prominent feature of DBA, the disease is also characterized by cancer predisposition, growth retardation and congenital malformations, in particular craniofacial, upper limb, heart and urinary system defects, which are present in ∼30-50% of patients. We completed our large scale sequencing of 80 ribosomal protein (RP) genes and found eight of them mutated in DBA. In total, together with three RP genes identified by others, there are 11 genes mutated in ∼54% of DBA patients; RPS19, RPS24, RPS17, RPL35A, RPL5, RPL11, RPS7, RPS10, RPS26, RPL19 and RPL26. To search for moderate and large RP gene deletions and duplications we performed high resolution array comparative genomic hybridization on 80 DNA samples from DBA patients who did not have mutations in the 11 known RP genes. We found a deletion of exon 2 and 3 (4800 bp), deletion of the coding region, and duplication of exons 2 and 3 (488 bp) in RPS19 gene in three probands; three deletions of exons 1, 2 and 3 in RPS17 in three probands (2920 bp, 2886 bp and 3018 bp); and deletion of exons 1, 2 and 3 of the RPS26 gene. We also identified two deletions and a duplication in three RP genes previously not found mutated in DBA; RPS8 duplication of exon 3 (764 bp), RPS14 deletion of exons 2, 3, 4 and 5 (2568 bp) and RPS15 deletion of exon 4 (1995 bp). The deletions and duplications are being confirmed by multiplex PCRs. Interestingly, RPS14 was previously identified as a 5q- syndrome gene demonstrating that abnormality of this protein can cause both DBA and 5q- syndrome. These data bring to 14 the total number of RP genes mutated in DBA.

Disclosures:
No relevant conflicts of interest to declare.
Phase II Study of the Combination of Clofarabine Plus Idarubicin and Cytarabine (CIA) In Patients with Relapsed/Refractory Acute Myeloid Leukemia (AML).
Authors: Naqvi Kiran , Ravandi Farhad , Kantarjian Hagop , Borthakur Gautam , Ferrajoli Alessandra , Thomas Deborah A , Koller Charles Asa , Konopleva Marina , Byrd Anna , Kwari Monica , Faderl Stefan 
Published: October 2019

Abstract
Abstract 1061
Clofarabine is a second generation nucleoside analog with activity in patients with acute leukemias. Previous studies with the combination of clofarabine plus cytarabine have reported a complete remission rate (CR) of 24% in relapsed AML and 52% in previously untreated AML patients. To explore further combinations with drugs active in AML, we conducted a phase II study of CIA in patients with relapsed and primary refractory AML. Patients were treated at the maximum tolerated dose (MTD) determined from a preceding phase I study: clofarabine 22.5mg/m2 IV daily × 5, idarubicin 6 mg/m2 IV daily × 3, and cytarabine 0.75 g/m2 IV daily × 5 days. A total of 63 patients were enrolled. Patients with prior exposure to clofarabine were not eligible. The median age was 51years (18 - 80years). Twenty-four patients were primary refractory and 39 had relapsed disease. All the patients received CIA in 1st salvage. For patients with relapsed AML, the median duration of the first CR was 36 weeks (3 – 215 weeks). Three patients underwent stem cell transplant (SCT) prior to treatment with CIA and 28 patients had received at least intermediate doses of cytarabine prior to relapse. Fifteen patients had secondary AML following a preceding phase of MDS. Twenty-five percent of patients had -5/-7 and 6% had trisomy 8. Thirteen patients (21%) achieved CR, 7 (11%) CRp (complete remission without platelet recovery), and 4 (6%) a partial remission (PR) for an overall response rate of 38%. The response rate in patients with primary refractory disease was 33% while the response rate in patients with relapsed disease was 41%. Five patients (8%) died during induction (</= 28 days); 4 from septic shock and 1 from diffuse alveolar hemorrhage. Median duration of follow-up was 65 weeks (range 6–161). Median overall survival was 34 weeks (range 1–161). Disease free survival for those patients who achieved CR/CRp was 66 weeks (range 4–156). A total of 24 (38%) patients underwent SCT following CIA. The most common toxicity was nausea/vomiting in 49 (78%) patients followed by hepatic dysfunction in 43 (68%) patients and diarrhea in 42 (67%) patients. In summary, the CIA combination has activity in patients with relapsed/refractory AML. Further evaluation in better prognosis patients is ongoing.

Disclosures:
Ravandi: Genzyme: Honoraria, Membership on an entity's Board of Directors or advisory committees. Kantarjian:Genzyme: Research Funding. Faderl:Genzyme: Research Funding.
Phase II Study of All-Trans Retinoic Acid (ATRA), Arsenic Trioxide (ATO), with or without Gemtuzumab OzogamIcin (GO) for the Frontline Therapy of Patients with Acute Promyelocytic Leukemia (APL).
Authors: Ravandi Farhad , Estey Elihu H. , Cortes Jorge E. , O'Brien Susan , Pierce Sherry A. , Brandt Mark , Ferrajoli Alessandra , Borthakur Gautam , Konopleva Marina , Verstovsek Srdan , Garcia-Manero Guillermo , Faderl Stefan , Kantarjian Hagop 
Published: October 2019

Abstract
Abstract 1080

Background:
The role of arsenic trioxide (ATO) in the frontline treatment of patients with acute promyelocytic leukemia (APL) remains unclear with a number of studies reporting high and durable responses with single agent ATO. We have conducted a trial combining all-trans-retinoic acid (ATRA) with ATO with or without gemtuzumab ozogamicin (GO) in patients with previously untreated APL.

Patients and methods:
From July 2002 to June 2010, 104 patients with newly diagnosed APL were treated with a combination of ATRA plus ATO in two studies. The first cohort of 47 patients received ATRA (45 mg/m2 daily) and ATO (0.15 mg/kg daily beginning on day 10 of ATRA). High-risk patients (White blood cell count [WBC] > 10 × 109/L) received GO 9 mg/m2 on the first day of induction. From July 2007, the second cohort of 57 patients received ATRA (45 mg/m2 daily) and ATO (0.15 mg/kg daily) concomitantly on day one of induction. They also received GO 9 mg/m2 on day 1, if high risk, and any time during induction if the WBC rose to > 30 × 109/L (and more recently if > 10 × 109/L). Monitoring for PML-RARA fusion gene using reverse transcriptase-polymerase chain reaction (RT-PCR) was conducted after induction and throughout consolidation and follow up. The median age for the 104 patients was 46 years (range, 14–81). Their median presenting WBC was 2.7 × 109/L (0.4-131.4 × 109/L) and their median platelet count was 36 × 109/L (range, 7–261 × 109/L). Seventy three (70%) had low risk and 31 (30%) high risk disease (based only on the presentation WBC ≤ or > 10.0 × 109/L).

Results:
Overall, 102 patients (98%) achieved complete remission (CR) and 2 died during induction. With a median follow-up of 115 weeks (range, 4 to 397 weeks), 94 patients remain alive. The estimated 5-year survival rate was 88% and event-free survival 86%; only 5 of the patients achieving a CR (5%) have relapsed. The median overall survival, remission duration and event-free survival have not been reached (Figure). Thirty six patients have been alive and in remission for more than 3 years and 21 for more than 5 years. Two late deaths (beyond 300 weeks) occurred in CR from unrelated causes.

Conclusion:
The combination of ATRA and ATO (with or without GO) as initial therapy for APL is highly effective and safe; it can potentially substitute for chemotherapy containing regimens in high and low risk patients.

Disclosures:
Ravandi: Cephalon: Honoraria, Research Funding, Speakers Bureau. Off Label Use: Off-label use of arsenic trioxide in frontline therapy of APL; off label use of gemtuzumab ozogamicin in APL. Verstovsek:Incyte Corporation: Research Funding.
Rapid One-Tube Long PCR-Sequencing for ABO Genotyping From Exons 2 to 7.
Authors: Park Geon , Jang Sook Jin , Moon Dae Soo , Park Young Jin , Huh Ji Young 
Published: October 2019

Abstract
Abstract 1116

Background:
ABO is the most clinically important blood group system in transfusion and transplantation medicine. DNA-based methods for determining the ABO genotype have markedly advanced in recent years and have been increasingly introduced in clinical laboratories. Most studies using sequencing methods for ABO genotyping have focused on exons 6 and 7, ignoring the other exon and intron sequences. Although these approaches usually can distinguish among common ABO alleles, erroneous or ambiguous typing results might be obtained in cases of hybrid and subgroup alleles with various missense or nonsense mutations. For more accurate ABO genotype results, the entire exon sequence including the introns and regulatory regions should be analyzed, but such sequencing methods are time-consuming, labor intensive, and expensive.

Methods:
To investigate the entire sequence of ABO exons 2–7 including introns 2–6 and excluding non-coding region of exon 7, long shuttle one-tube PCR-sequencing method was designed. We used a rapid DNA polymerase with high fidelity (Phusion® Flash II DNA Polymerase; Finnzyme OY, Espoo, Finland). ABO genotyping was performed using this technique in peripheral blood of two unrelated families.

Results:
Time requirement of the PCR amplification and purification processes were about 1.8 h and 20 min, respectively. Also, this technique could read the entirety of exons 2–7 including introns, required one PCR tube, one purification step, and 27 sequencing reactions for the entire sequencing. A total of seven different ABO alleles from two families were analyzed. All genotyping results agreed with serologic findings and results expected by Mendelian inheritance. The ABO phenotypes (genotypes) of family A members were A1 (A105/A105), O (O01/O02), A1 (A105/O01), A1 (A105/O02), and A1 (A105/O02) in the husband, wife, and three sons, respectively. The ABO genotype of husband in family B was cis-ABO1/O01 and his serologic ABO typing result was ABweak. His wife had the common AB phenotype (A1B); however, her genotype revealed a hybrid of A and B alleles and the B101 allele. The hybrid allele comprised the consensus ABO allele in exons 2–5 (including introns 2–4), the B101 allele in exon 6 (with introns 5 and 6), and the Al01 allele in exon 7.

Conclusion:
This method is suitable for the fast analysis of ABO genotypes and may be valuable in clinical transfusion or forensic applications.

Disclosures:
No relevant conflicts of interest to declare.
FXII Antisense Oligonucleotide Mediated Depletion Results In Effective Anticoagulation without Bleeding Risk.
Authors: Crosby Jeff R. , Gao Dacao , Zhao Chenguang , May Chris , MacLeod Robert , Monia Brett P. 
Published: October 2019

Abstract
Abstract 1149
Human clinical efficacy has been demonstrated with antisense oligonucleotides (ASOs) in several disease indications, including dyslipidemia, cancer, diabetes and multiple sclerosis. Coagulation factors are attractive targets for antisense therapeutics for several reasons. The liver is principally responsible for production of coagulation factors and is an organ which is highly sensitive to ASO drugs. ASOs are highly specific, and thus we can selectively evaluate multiple coagulation factors to determine their roles in thrombosis, bleeding, and other processes. Employing antisense technology we selectively reduced levels of several individual clotting factors and compared the relative risk/benefit profiles. Antithrombotic activity was determined following ferric chloride induced thrombosis in the inferior vena cava and bleeding tendency was measured by blood volume loss following tail nick. ASOs targeting several clotting factors were studied, including factors FII, FVII, FIX, FXI and FXII. Previously we characterized the beneficial effects of targeting FXI, demonstrating antithrombotic effects without increased bleeding risk. Here, we describe the effects of targeting another member of the intrinsic pathway, FXII. FXII ASO treatment resulted in a dose-dependent and specific reduction in FXII mRNA levels in liver. These reductions in FXII levels correlated well with a prolongation of aPTT with no effects on PT prolongation. The anticoagulant effect of FXII ASO treatment also correlated well with antithrombotic activity in a FeCl3 induced IVC thrombosis mouse model across a wide dose range. In addition to the venous thrombosis model, FXII ASO treatment was effective across a wide dose range in a model of arterial thrombosis. Furthermore, FXII ASO treatment was well tolerated and no prolongation of tail bleeding time was observed at any dose tested, indicating a broad safety margin for FXII ASO targeting (in mice). In addition, FXII inhibition was protective in a mouse TF driven pulmonary embolism model. Inhibiting coagulation factors with ASOs is highly specific and offers an attractive approach to identifying optimal antithrombotic targets as well as human therapeutics for the treatment of coagulation related disorders with the potential for improved safety profiles.

Disclosures:
No relevant conflicts of interest to declare.
Association of Interferon-Gamma +874(T/A) Single Nucleotide Polymorphism with Response to Immunosuppressive Therapy In Patients with Severe Aplastic Anemia.
Authors: Chang Hong , Zhang Jia-You , Mu Xiao-Yan , Meng Wen-Tong , Liu Ting 
Published: October 2019

Abstract
Abstract 1157
The involvement of IFN-γ in the pathophysiology of AA has been well documented, and overexpression of IFN-γ in the marrow and blood of patients with AA has been reported. T to A single nucleotide polymorphism (SNP) at position +874 of intron 1 of the IFN-γ gene (IFN-γ +874) can directly affect the expression of the IFN-γ gene since position +874 of intron 1 coincides with a putative nuclear factor (NF)-κB binding site. Thus far, no study has reported the correlation between the IFN-γ gene polymorphism and responsiveness of patients with SAA to immunosuppressive therapy. Therefore, we investigated the distribution of the IFN-γ +874 T/A gene by amplification refractory mutation system-polymerase chain reaction (ARMS-PCR), and analyzed the correlation between the IFN-γ +874 T/A gene and responsiveness of patients with SAA to IST. ARMS-PCR was used to amplify the polymorphic segments of the IFN-γ +874 T/A gene in the samples obtained from 54 patients with aplastic anemia and 51 healthy adults. Further, enzyme linked immunosorbent assay was used to assay IFN-γ levels in the blood plasma of 35 patients with severe aplastic anemia (SAA) before IST and 20 healthy blood donors. The results showed that the frequencies of IFN-γ +874 TT genotype in patients with aplastic anemia were significantly higher than the corresponding frequencies in the healthy adults (42.6% vs. 17.6%, χ2 = 13.780, p = 0. 01) (Table 1). The response rate in SAA patients with increased IFN-γ levels in the blood plasma was higher than that in SAA patients with decreased IFN-γ levels in the blood plasma (73.7% vs. 25.0%, p < 0.05). Of the 35 patients with SAA, 15 showed the IFN-γ +874 TT genotype, whereas response in 11 patients, the high response rate was significantly in the favor of the IFN-γ +874 TT genotype (73.3% vs. IFN-γ +874 non-TT 35%, p < 0.05) (Table 2). In conclusion, the IFN-γ +874 T/A gene polymorphism may be correlated with response to immunosuppressive therapy. Our study showed that the frequencies of IFN-γ +874 TT genotype in patients with AA were significantly higher than the corresponding frequencies in the healthy adults, thereby suggesting that IFN-γ +874 A/T gene polymorphism is correlated with the susceptibility of aplastic anemia. Our study also showed that a high concentration of IFN-γ in the blood plasma before receiving IST and IFN-γ +874 TT genotype were predictors of a good response to IST; this finding was consistent with that of other studies.

Disclosures:
No relevant conflicts of interest to declare.
The Transcription Factor ELF4 Promotes Survival of Myeloid Leukemic Stem Cells.
Authors: Park Chun Shik , Suppipat Koramit , Lacorazza H. Daniel 
Published: October 2019

Abstract
Abstract 1209
Chronic myeloid leukemia (CML) is a myeloproliferative disease that originate in hematopoietic stem cells (HSCs) as a result of the t(9;22) translocation, giving rise to the Ph (Philadelphia chromosome) and BCR-ABL oncoprotein. Although treatment of CML patients with tyrosine kinase inhibitor can efficiently eliminate most leukemic cells, chemoresistant leukemic stem cells (LSCs) can survive and drive recurrence of CML in these patients. A number of genes have been described to promote or inhibit proliferation of LSCs. Some of them have similar roles in normal HSCs. The transcription factor ELF4 promotes cell cycle entry of quiescent HSCs during homeostasis (Lacorazza et al., 2006). Thus, to investigate the function of ELF4 in CML initiation and maintenance, we developed a BCR-ABL-induced CML-like disease using retroviral transfer of BCR-ABL in Elf4-null bone marrow (BM) cells. We first investigated whether ELF4 is required for the induction of CML. Recipient mice of BCR-ABL-transduced WT BM cells developed CML and died with a latency 16–23 days, whereas recipient mice of BCR-ABL-transduced Elf4-/- BM cells showed longer latency of 45–47 days (n=20; p<0.0005). Progression of leukemia was monitored in peripheral blood, BM and spleen by flow cytometry. In mice transplanted with BCR-ABL-transduced Elf4-null BM cells, Gr-1+ leukemic cells expanded the first two weeks after BM transplantation followed by a decline at expense of a secondary expansion of B220+ cells. In contrast, Gr-1+ leukemic cells continuously expanded in mice receiving BCR-ABL-transduced WT BM cells. These results suggest that loss of ELF4 causes a profound abrogation in BCR-ABL-induced CML, while allowing progression of B-cell acute lymphocytic leukemia. Since loss of Elf4 led to impaired maintenance of myeloid leukemic cells, we postulated that ELF4 may affect survival of LSCs. Thus, we analyzed the frequency of Lin-c-Kit+Sca-1+ (LSK) cells that are BCR-ABL positive in BM and spleen. We found that BCR-ABL+ LSK cells were significantly reduced in recipients of BCR-ABL-transduced Elf4-/- BM cells. These studies indicate that ELF4 is essential to maintain the LSC pool in CML acting as a molecular switch between myeloid and lymphoid blast crisis.

Disclosures:
No relevant conflicts of interest to declare.
The Gamma Catenin/CBP Complex Maintains Survivin Transcription In β-Catenin Deficient/Depleted Cancer Cells.
Authors: Kim Yong-Mi , Ma Hong , Oehler Vivian , Jiang Enzi , Nguyen Cu , Masiello David P. , Liu Han , Zhao Yi , Radich Jerald P. , Kahn Michael 
Published: October 2019

Abstract
Abstract 1218
Survivin is a member of the inhibitor of apoptosis (IAP) gene family, which plays essential roles in prevention of apoptosis, cell division, and cancer development. Aberrant expression of Survivin is associated with essentially all tumors compared to their normal tissues. A number of signaling pathways are involved in the transcriptional regulation of Survivin. Previously, we demonstrated that Survivin expression is CBP/β-catenin/TCF-dependent and that ICG-001, a specific inhibitor of binding to the N-terminus of CBP, effectively attenuates Survivin expression. Now, using NCI-H28 cells, which harbor a homozygous deletion of β-catenin, we demonstrate that Survivin transcription can similarly be mediated by nuclear γ-catenin. In this study, we confirm that γ-catenin, like β-catenin, accumulates in the nucleus and that it can increase TOPFLASH reporter activity upon Wnt3a stimulation in NCI-H28 cells. In addition, overexpression of γ-catenin increases the expression of the TCF/catenin target genes Survivin and cyclinD1. This effect can be diminished by overexpression of dominant negative TCF or transactivation domain deleted γ-catenin. We demonstrate that g-catenin by binding to TCF family members and specifically recruiting the coactivator CBP drives Survivin transcription particularly in β-catenin-deficient cells indicating that γ-catenin can participate in the canonical Wnt signaling cascade. We also examined the relative expression of γ-catenin and β-catenin in 90 cases of chronic myeloid leukemia (CML) in a published gene expression microarray data base. A statistically significant negative correlation between γ-catenin and β-catenin was found in AP/BC cases (-0.389, P = 0.006). Furthermore, in subsequent independent validation studies by qPCR in 28 CP and BC patients increased g-catenin expression predominated in BC cases and was associated with concomitantly increased Survivin expression. Gene expression was 3- and 6-fold greater in BC patients as compared to CP patients, for γ-catenin and survivin, respectively. Consistent with this observation, nuclear γ-catenin accumulation was evident in this population consistent with a potential transcriptional role. Combined treatment with imatinib mesylate (IM) and ICG-001 significantly inhibited colony formation in sorted CD34+ CML progenitors (survivin+/γ-cateninhigh/β-cateninlow) isolated from one BC and one AP patient resistant to IM. Our data indicates the potential of ICG-001 to block both the CBP/γ-catenin interaction and the CBP/β-catenin interaction and this may have clinical significance in cancers in which γ-catenin plays a significant transcriptional role.

Disclosures:
No relevant conflicts of interest to declare.