bluuebunny/crossref_metadata_2025_split huggingface dataset:
Development Of Novel Alkylating Chemotherapeutics, Phenyl-Chloroethyl Urea Family, Targeting Mitochondrial Prohibitin For Selective Killing Of Leukemic Cells
Authors: Kim Hye-Ran , Kim Hwan-Young , Thi Dai Trang Nguyen , Lee Il-Kwon , Kim Hyeoung-Joon , Shin Jong-Hee , Suh Soon-Pal , Ryang Dong-Wook , Shin Myung-Geun 
Published: October 2019

Abstract

Background
Mitochondrial aberrations have been associated with chronic degenerative diseases, aging and cancer. Mitochondrial proteins are interesting targets for the development of selective anticancer drugs in leukemia and other malignancies. We analyzed mitochondrial proteins to develop novel anti-cancer agents targeting selective leukemic cell, especially acute myelocytic leukemia (AML) cells.

Materials and Methods
Mitochondria were isolated from primary AML cells and AML cell lines (THP-1 and KG-1) by density-gradient ultracentrifugation using swelling buffer and sucrose buffer. Dysregulated mitochondrial protein were identified using 2-DE and mass spectrometry (MALDI-TOF/TOF technology). For in vivo experiments, AML cell grafts were established in 6-week-old Balb/c mice by subcutaneous injection of 1x107 THP-1 AML cells in the right flank at day 0.

Results
Out of these deregulated proteins, totally 12 and 20 proteins were observed in up- or down-regulated spots, respectively. Interestingly, prohibitin (gi4505773) was highly expressed in all type of leukemic cells, especially primary AML cells (Fig. 1), which is confirmed by immunoblot and immunofluorecenct methods. Aberrant expressed prohibitin induced growth suppression and repressed E2F-mediated transcription. We synthesized potent chemical substances that can alkylate PHB, two molecules of phenyl-chloroethyl urea family such as cyclohexylphenyl-chloroethyl urea (CCEU) and iodophenyl-chloroethyl urea (ICEU) from the reaction with 2-ethylisocyanate and 4-cyclohexylaniline and 4-iodoaniline, respectively. They were characterized by 1H NMR and 13C NMR. Time and dose dependent manner of proliferation suppression when treated with CCEU and ICEU was observed in primary AML and AML cell lines. Notably morphological transformation of AML cells was observed when treated with 10 - 100 umol of CCEU and ICEU for 24 hours. The half maximal inhibitory concentration (IC50) was 25umol of most AML cell lines. Cell cycle analysis of CCEU-and-ICEU-treated- AML cells showed a remarkable increase of the sub-G1 phase. Immunoblotting experiment revealed the change of cytoplasmic and nucleoplasmic PHB. The increment expression of cleaved caspase3 and poly ADP-ribose polymerases were also observed in AML cell lines. Moreover, CCEU and ICEU selectively killed AML cells in mouse model (Fig. 2).

Conclusion
This study showed the development of novel alkylating chemotherapeutics, phenyl-chloroethyl urea family (CCEU and ICEU), targeting mitochondrial prohibitin for selective killing of leukemic cells. Selective eradication effects of these novel chemotherapeutics were further confirmed using in vivo mice model.

Disclosures:
No relevant conflicts of interest to declare.
Second Hematologic Malignancies After Radioiodine Exposure In Patients With Thyroid Cancer and The Relationship With Radioiodine Dosage: A Nationwide Population-Based Study
Authors: Liu Chia-Jen , Hung Man-Hsin , Teng Chung-Jen , Hu Yu-Wen , Hung Yi-Ping , Gau Jyh-Pyng , Chiou Tzeon-Jye , Tzeng Cheng-Hwai 
Published: October 2019

Abstract

Background
Second primary malignancies are one of the most important late effects after radioactive iodine (RAI) exposure, which is widely used in the diagnosis and treatment for thyroid cancers. However, dose-response correlation between RAI and second hematologic malignancies (SHM) was not established. Therefore, we conducted a nationwide population-based study to investigate the risk of SHM among thyroid cancer patients and the association between RAI dosage and development of SHM, including leukemia.

Material and methods
We recruited patients with newly diagnosed thyroid cancer aged 20 years or older without antecedent cancer from the Taiwan National Health Insurance database between 1997 and 2010. The standardized incidence ratios (SIR) of cancers were calculated to compare the cancer incidence of thyroid cancer patients to the general population according to gender, calendar year, and age in 5-year intervals by the corresponding stratum-specific person-time accrued in the cohort. The association of cumulative RAI dosage and development of SHM, including leukemia, was estimated using time-dependent analysis with 2-year latent period.

Results
After exclusion of first-year follow-up, 692 second primary malignancies developed among 20,235 patients with thyroid cancer, with a follow-up of 134,178 person-years. Fifty SHM developed with a SIR of 2.37 (95% confidence interval 1.76-3.13). The significant increased risk of SHM included: leukemia (SIR, 2.74; 95% CI, 1.65-4.28), non-Hodgkin lymphoma (SIR, 2.38; 95% CI, 1.55-3.48). As a time-dependent covariate with a 2-year latent period, RAI significantly increased the risk of leukemia with a clear dose-response correlation (age- and sex-adjusted hazard ratio per 100 mCi increase, 1.10; 95% CI, 1.01-1.19, P = 0.032). This effect was not seen in other second primary malignancies.

Conclusions
Our study reveals increased risk of SHM among patients with thyroid cancer. RAI increases the risk of leukemia with a clear dose-response relationship.

Disclosures:
No relevant conflicts of interest to declare.
Genomic Heterogeneity In Pediatric Acute Lymphoblastic Leukemia
Authors: Yaniv Isaac , Jeison Marta , Hameiri-Grossman Michal , Mardoukh Jacque , Sharon Nir Adam , Elitzur Sarah , Gilad Gil , Avrahami Gali , Stark Batia , Avigad Smadar 
Published: October 2019

Abstract
Pediatric B cell Precursor Acute Lymphoblastic Leukemia (BCP-ALL) is characterized by recurrent chromosomal aneuploidies and translocations, which are important for diagnosis and risk stratification. Classical cytogenetic and FISH are routinely used for the diagnosis of most of these alterations, particularly those involving balanced translocations, and numerical changes. Recently, genome wide analysis, such as aCGH and SNP technologies have identified copy number alterations (CNA) in genes involved in lymphoid differentiation: IKZF1, PAX5, EBF1 BTG1; and in cell cycle control: CDNK2A/B, RB1 and ETV6. Multiplex ligation–dependent probe amplification (MLPA) technique has been used for the identification of those focal genetic aberrations in a single assay but it cannot reliably detect aberrations present in less than 50% of cells. Cytoscan HD SNP array (Affymetrix) can overcome this limitation and is the only array which enables independent confirmation of CNA with SNP allelic information. Thus minor genetic subclones missed by other methods, can be detected.
The aim of this work was to compare between SNP array and MLPA platforms in a routine diagnostic setting.
Samples of 39 pediatric BCP-ALL patients well characterized by classical cytogenetic and FISH at diagnosis, were studied by MLPA P335 (MRCHolland) and SNP array (Affymetrix). In total 273 genomic loci were evaluated, 13 (5%) were not informative, of them 7 were not covered by the MLPA kit and 6 gave not evaluable MLPA results. 231/260 (89%) of the loci showed concordance, 39/260 (15%) with CNA and 192/260 (74%) normal. A total of 68 CNA were detected, 29 were discordant and 39 concordant.
IKZF1 deletion was identified by both methods in 5 cases (13%). However, a deletion was identified in 3 additional samples, only by cytoscan. In these cases, the deletion was present in a minor subpopulation (in 45% or less of the cells), while the main cytogenetic aberration was present in the majority of cells (60%-90%).
We conclude that although MLPA is a useful technique, it is not sensitive enough to detect small populations. In this small cohort, 3 IKZF1 deletions were missed since they were present only in a minor subclone. Interestingly, one of these patients relapsed, and the relapse clone harbored the IKZF1 deletion.

Conclusion
Genome-wide sensitive methods identify different subpopulations in each patient, thus detecting previously unrecognized genetic heterogeneities.

Disclosures:
No relevant conflicts of interest to declare.
The Combination Of 2-DG and Metformin Inhibits The mTORC1 Pathway and Suppresses Aggressive B Cell Lymphoma Growth and Survival
Authors: Bi Chengfeng , Fu Kai , Jiang Chunsun , Huang Xin , Chan Wing Chung , McKeithan Timothy 
Published: October 2019

Abstract
mTORC1 (mammalian target of rapamycin complex 1) plays a central role in integrating nutrient and growth factor inputs to control cell growth in all eukaryotes and is commonly deregulated in human cancers. Inhibition of mTORC1 is a promising strategy in lymphoma therapy. However, only a few drugs, such as rapamycin and its analogs (rapalogs), have been approved for treatment in a limited number of cancer types, due to their incomplete and nonspecific inhibition of mTORC1 as well as their limited effects toward the 4EBP1 pathway. 4EBP1 pathway regulates protein translation which is considered to be crucial in cancer cell survival and proliferation. In this study, we used the glycolysis inhibitor 2-deoxyglucose (2-DG) together with the mitochondrial respiratory inhibitor metformin to treat aggressive B cell lymphoma cells in vitro and in vivo. We found that the combined treatment inhibited mTORC1 and its major downstream targets, including 4EBP1. As a result, combined treatment significantly inhibited tumor cell growth and survival by the inhibition of 5’ cap-dependent translation involving lymphoma associated oncogenes such as MCL-1, BCL-XL and Cyclin D1. Moreover, the combination of 2-DG and metformin suppressed tumor growth in B cell lymphoma xenograft mouse models. Although the combined treatment dramatically decreased cellular ATP levels, mTORC1 inhibition was independent of AMPK activity but instead resulted from inhibitory effects on Rag-GTPases, which are upstream activators of mTORC1. Given that both 2-DG and metformin have been used in clinical diagnosis or treatment for decades, the combination of the two drugs hold promise as a new strategy to treat aggressive B cell lymphoma.

Disclosures:
No relevant conflicts of interest to declare.
Excessive Early Mortality Drives Poor Long Term Outcomes Among Older Patients With Classical Hodgkin Lymphoma: A Population Study
Authors: Costa Luciano J. 
Published: October 2019

Background
Classical Hodgkin Lymphoma (cHL) is a highly curable malignancy in young adults. Most of the information available on natural history and management of cHL comes from series and clinical trials in young patients. There is limited information on the outcomes of older patients with cHL

Methods
We analyzed the characteristics and outcomes of a large contemporary cohort of cHL patients ≥ 65 years diagnosed in the US and reported to the Surveillance Epidemiology and End Results program (SEER-18). Inclusion criteria consisted of diagnosis of cHL as first malignant neoplasm, year of diagnosis 2000-2010, known stage and known race. Cases reported from death certificate or autopsy only were excluded. Information retrieved contained age at diagnosis, year of diagnosis, race, sex, histological subtype, stage, presence of extra-nodal disease and survival time. Characteristics of cHL in patients ≥ 65 years (older) were compared to those < 65 years (younger). We subsequently estimated relative survival (RS) for different stages and age strata among older patients comparing with outcomes in younger patients.

Results
There were 20815 cases of cHL reported during the period with median follow up of 48 months, including 2884 (13.8%) cases in older patients. Older patients were more likely to present with extra-nodal disease (4.7% vs. 2.2%, P< 0.001), advanced stage (52.6% vs. 37.3%, P<0.001) and with lymphocyte depleted cHL (3.3% vs. 0.9%) or mixed cellularity cHL (22.5% vs. 11.3%, P<0.001) than younger patients. Additionally, older cHL patients with early stage disease were less likely to receive radiation therapy (35.6% vs. 48.2%, P<0.001). RS at one year (64% vs. 95.7%, P<001) and 5-years (49% vs. 89%, P<0.001) was much inferior in older than in younger patients, even when stratified by early (I and II) and advanced (III and IV) stages (Figure). RS deteriorated quickly with increasing age at diagnosis. One year RS was 73.9%, 56.3% and 43.3% and 5-years RS was 57.9%, 41.6% and 30.4% for patients 65-74, 75-84 and 85+ years, respectively. Even with adjustment for histology, sex, year of diagnosis and race, patients 65-74 years (HR = 4.81, 95%CI 4.41-5.26, P<0.001) 75-84 years (HR = 9.60, 95% CI 8.78-10.49, P<0.001) and 85+ years (HR = 16.19, 95% CI 14.15-18.51, P<0.001) were at a much higher risk of death than younger cHL patients.

Conclusion
cHL has distinct presentation and far worse prognosis in older patients with excessive early (<1 year) mortality. There is a need to further understand patterns of treatment failure and develop age-specific therapeutic interventions for this group.

Disclosures:
No relevant conflicts of interest to declare.
In Vivo Disruption Of The Hepcidin−Ferroportin Regulatory Circuitry Causes Fatal Systemic and Exocrine Pancreatic Iron Overload
Authors: Altamura Sandro , Gröne Hermann Josef , Kessler Regina , Galy Bruno , Hentze Matthias W. , Muckenthaler Martina U. 
Published: October 2019

Abstract
Systemic iron levels are tightly controlled by the hepatic hormone hepcidin in response to iron availability, inflammation, hypoxia or the iron demand for erythropoiesis. Hepcidin binds to the iron export protein ferroportin (FPN1) to regulate iron release from exporting cells. A mutation of cysteine 326 (C326S) of FPN1 was reported in a patient with non−classical ferroportin disease (Sham et al, 2005) and shown to abrogate hepcidin binding in vitro (Fernandes et al, 2009).
To study consequences of the disruption of the hepcidin−ferroportin interaction in vivo, we generated the first knock−in mouse model of C326S non−classical ferroportin disease.
Mice with either heterozygous or homozygous C326S FPN alleles are viable and fertile. At 8−weeks of age both heterozygous and homozygous mice show profoundly increased transferrin saturation and serum ferritin levels as well as hepatic iron overload. Histological analysis by Perl’s Prussian blue staining revealed that hepatic iron accumulation is restricted to hepatocytes and that Kupffer cells are spared of iron. In addition, splenic macrophages and duodenal enterocytes are iron−depleted.
Macroscopically, C326S homozygous mice show progressive, brown discoloration of the pancreas that correlates with profound iron deposition. Histological analysis reveals that iron localizes exclusively to the exocrine pancreas sparing the islets of Langerhans. Consistently, C326S homozygous mice do not show any signs of diabetes. Pancreatic iron accumulation is closely associated with increased reactive oxygen species (ROS), degeneration of exocrine pancreatic cells, increased plasma lipase and exocrine pancreatic failure. Starting at the age of 33 weeks, pancreatic failure is accompanied by progressive wasting and death. We believe that C326S FPN mice represent the first example of fatal iron overload in an animal model, opening avenues to investigate the underlying molecular mechanisms.
Sham R, Phatak PD, West C, et al. Autosomal dominant hereditary hemochromatosis associated with a novel ferroportin mutation and unique clinical features. Blood Cells Mol. Dis. 2005; 34:157−61.
Fernandes A, Preza GC, Phung Y, et al. The molecular basis of hepcidin−resistant hereditary hemochromatosis. Blood. 2009;114:437−443.

Disclosures:
No relevant conflicts of interest to declare.
Apixaban Reduces Hospitalization In Patients With Venous Thromboembolism: An Analysis Of The AMPLIFY-EXT Trial
Authors: Liu Xianchen , Thompson John , Phatak Hemant , Mardekian Jack , Porcari Anthony R. , Johnson Margot R. 
Published: October 2019

Abstract

Introduction
Venous thromboembolism (VTE) is associated with a considerable risk for morbidity and recurrence and related hospitalizations. In the Apixaban after the Initial Management of Pulmonary Embolism and Deep Vein Thrombosis with First-Line Therapy-Extended Treatment (AMPLIFY-EXT) trial, a double-blind placebo-controlled trial with 12 months of treatment, two doses of apixaban (2.5 mg and 5 mg, twice daily) versus placebo significantly reduced symptomatic recurrent VTE or all-cause death without increasing the rate of major bleeding among 2,482 VTE patients who had completed 6-12 months of anticoagulation therapy. In this study, the effects of apixaban therapy versus placebo on medical hospitalization during AMPLIFY-EXT trial were evaluated.

Methods
A total of 2,477 patients who received study drugs were included in the analysis. All-cause hospitalizations during the trial were captured by dedicated case report forms. Outcomes of interest were; rate of hospitalizations and time from randomization to the first hospitalization. Patients were censored at either death, loss to follow-up, or end of study, whichever came first. Effects of treatment with apixaban versus placebo on the rates of hospitalization were assessed using Cox proportional hazards regression models.

Results
During a mean follow-up of 12.3 months, 138 patients were hospitalized at least once, 62 (7.5%/year) in the placebo group (n=826), 42 (4.8%/year) in the apixaban 2.5 mg group (n=840), and 34 (4.0%/year) in the apixaban 5 mg group (n=811). Compared with placebo, apixaban 2.5 mg [hazard ratio (HR) 0.65, 95% confidence interval (CI) 0.44–0.96; p=0.030] and 5 mg (HR 0.54, 95%CI 0.36–0.83, p=0.004) were both associated with significant reduction in hospitalization. There was no significant difference in hospitalizations between the 2 doses of apixaban (5 mg vs. 2.5 mg: HR 0.84, 95%CI 0.53–1.32, p=0 .445). The mean time to first hospitalization was 153.7 days in the placebo group, 196.9 days in the apixaban 2.5 mg group, and 202.4 days in the apixaban 5 mg group (Figure).

Conclusions
Extended anticoagulation with apixaban at either a dose of 5 mg or 2.5 mg significantly reduced the risk of hospitalization, possibly due to the reduction in VTE recurrence.

Disclosures:
Liu: Pfizer: Employment, Equity Ownership. Thompson:Pfizer: Employment, Equity Ownership. Phatak:BMS: Employment, Equity Ownership. Mardekian:Pfizer: Employment, Equity Ownership. Porcari:Pfizer: Employment, Equity Ownership. Johnson:Pfizer: Employment, Equity Ownership.
Thrombin Generation Measurement In Whole Blood As a Control For Vitamin K Antagonist Treatment - Effect Of Thrombomodulin
Authors: Bloemen Saartje , de Laat Marieke , Cate-Hoek Arina ten , Cate Hugo ten , De Laat Bas , Hemker H. Coenraad , Al Dieri Raed 
Published: October 2019

Abstract

Introduction
We tested whether the recently introduced measurement of thrombin generation (TG) in whole blood can be used to evaluate the clotting status of patients on vitamin K antagonist (VKA) prophylaxis. The prothrombin time, and hence the International Normalized Ratio (INR), only evaluates the vitamin K dependent factors II, VII and X but not the anticoagulant factors, protein C and S as well as factor IX. In TG all factors play their role and when thrombomodulin (TM) is added the function of proteins C and S is stressed. The thrombotic tendency in congenital protein C resistance proves the importance of this protein C pathway.

Aim
To compare the INR in samples from patients under VKA prophylaxis to TG in whole blood and in platelet rich and platelet poor plasma (PRP, PPP) both in the presence and in the absence of TM.

Materials & Methods
Blood samples were collected from 123 consenting patients on VKA. In two thirds (67%) the indication for prophylaxis was atrial fibrillation. Other indications included prosthetic valves, lung embolisms or thrombosis. The INR was determined in the PPP of the samples and the patients were stratified into 5 groups: INR of 1.0 to 1.5, 1.5 to 2.5, 2.5 to 3.5, 3.5 to 4.5 and higher than 4.5.
Thrombin generation (TG) was measured via Calibrated Automated Thrombinography (CAT) in whole blood and in PRP and PPP, with and without 20 nM added TM. From the TG curve lag time and time to peak were obtained as well as the maximal thrombin concentration (peak) and the area under the curve (endogenous thrombin potential: ETP). Also red and white blood cells and platelets were counted.

Results
With increasing INR values, the ETP and peak height decrease and lag time and time to peak prolong. All TG parameters measured in whole blood were significantly correlated (p-values< 0.01) with the values determined in both PRP and PPP. INR was linearly correlated with lag time and time to peak (p-value< 0.01), whereas for the concentration dependent parameters (ETP and peak height) the correlation with the INR was hyperbolical (p-value< 0.01).
In plasma, 20 nM TM causes a diminution of ETP and peak of 50-60 % in normals and in patients in the INR 1 – 1.5 group. At higher INR values inhibition is between 25 and 40%, independent of the INR value.
In whole blood, on the contrary, the same concentration of TM causes around 30 % of inhibition in normals and in all patients alike.

Conclusions
Whole blood TG data correlate well with INR and reflect more of the coagulation mechanism than the INR does. Like the INR it does not reflect the function of the VKAs on the natural anticoagulant factors, however. In PPP and PRP addition of TM shows that VKA treatment induces TM resistance in patients with an INR value higher than 1.5.

Disclosures:
No relevant conflicts of interest to declare.
Hematopoietic Cell Transplantation (HCT) For Fanconi Anemia (FA): Comparable Outcomes Regardless Of Stem Cell Source
Authors: MacMillan Margaret L. , Defor Todd E. , Wagner John E. 
Published: October 2019

Abstract

Historically, outcomes after alternative donor (AD)-HCT for FA have been strikingly inferior to those observed in recipients of HLA-matched sibling donor (MSD) hematopoietic stem cells (HSC) with excessive rates of graft failure, graft-versus-host-disease (GVHD), regimen related toxicity and infection, resulting in poor survival rates. Between 2006-2013, 44 FA patients with marrow aplasia and good organ function underwent AD-HCT after total body irradiation 300 cGy (single fraction) with thymic shielding, fludarabine (FLU) 140 mg/m2, cyclophosphamide (CY) 40 mg/kg and antithymocyte globulin (ATG). Outcomes were compared to those transplanted with HSC from an HLA matched sibling donor (n=24) after FLU 175 mg/m2, CY 20 mg/kg and ATG conditioning (1999-2013). GVHD preventative measures were identical with all recipients of marrow having the graft T cell depleted by CD34 selection (regardless of donor type) prior to infusion in addition to cyclosporine A and methylprednisolone or mycophenolate mofetil. Recipients of umbilical cord blood had no additional graft processing.
Except for higher use of pre-HCT G-CSF in recipients of AD-HCT, patient characteristics were similar between the 2 groups. Probabilities of neutrophil recovery, acute and chronic GVHD were similar between the groups (Table 1). Most notably, probability of survival at 3 years was the same between the two groups (Figure 1).
To our knowledge, this is the first demonstration of comparable outcomes after AD-HCT and MSD HCT for FA patients with marrow aplasia. These findings have three important implications: 1) timing for HCT need not be delayed if the patient lacks an HLA matched sibling donor, 2) potentially reduced enthusiasm for in vitro fertilization and preimplantation genetic diagnosis to have a 'savior sibling' and 3) ineligibility of FA young patients with a suitable AD (HLA matched adult volunteer or 5-6/6 matched UCB) and good organ function for high risk trials, including gene modified HSC.

Disclosures:
Wagner: Novartis: Research Funding.