bluuebunny/crossref_metadata_2025_split huggingface dataset:
Secondary Primary Cancers in Nodular Lymphocyte Predominant Hodgkin’s Lymphoma
Authors: Shameem Raji , Hamid Muhammad Saad , Hossain Nasheed Mohammad , Shani Dana 
Published: October 2019

Abstract
Background: Nodular Lymphocyte Predominant Hodgkin’s Lymphoma (NLPHL) is an uncommon subtype of Hodgkin’s Lymphoma. The risk for secondary primary cancers is not well understood.
Methods: The Surveillance Epidemiology and End Results (SEER) database was used to identify cases of NLPHL diagnosed up to 12/31/2011. The Standardized Incidence Ratio (SIR) was calculated as the ratio of observed to expected cases of secondary primary malignancies based on incidence data in the general United States population. The latency exclusion period from the date of diagnosis was 60 months. We also investigated for possible modifying effects of gender, age at diagnosis (20-39 years of age and ≥ 40 years of age), use of radiation therapy, and latency periods (5-10 years and >10 years) after initial diagnosis of NLPHL.
Results: A total 947 cases of NLPHL were present in the database. Secondary primary cancer cases occurred in 64 cases with 36 (56.3%) solid tumors and 28 (43.8%) hematological malignancy cases. Radiation was administered in 38 (59.4%) of secondary primary cancer cases. Females had an increased risk of Non-Hodgkin’s Lymphoma (NHL) (SIR: 13.86, p<0.05), “All Solid Tumors” (SIR: 1.89, p<0.05), Colon and Rectum (SIR: 4.39, p<0.05), and Urinary System (SIR: 2.01, p<0.05) tumors. Comparatively, males had an increased risk of NHL (SIR: 26.02, p<0.05) exclusively. The risk of developing NHL (SIR: 60.23, p<0.05), and “All Solid Tumors” (SIR: 2.47, p<0.05) was increased in individuals between 20-39 years of age. Whereas, increased risk of Urinary System (SIR: 3.78, p<0.05), and NHL (SIR: 12.59, p<0.05) were observed in the ≥ 40 years of age cohort. NHL (SIR: 21.0, p<0.05), “All Solid Tumors” (SIR: 1.65, p<0.05), and Colon and Rectum (SIR: 3.68, p<0.05) tumor risk was increased in cases treated with radiation therapy, while NHL (SIR: 21.0, p<0.05) was the only significantly increased secondary malignancy in non-radiated cases. Latency period of >10 years, after the initial diagnosis, was associated with increased risk of “All Solid Tumors” (SIR: 2.62, p<0.05), Thyroid cancer (SIR: 16.64, p<0.05), and NHL (SIR: 35.68, p<0.05).
Conclusion: NLPHL is associated with an increased risk of secondary primary cancers, both solid and hematological malignancies. Thyroid cancer and NHL were the most frequent tumors observed in patients ten years after initial NLPHL diagnosis. Radiation treatment for NLPHL increased the risk for solid tumors, including colorectal cancer. Further research is necessary to clarify optimal surveillance technique in patients after treatment of NLPHL.

Disclosures
No relevant conflicts of interest to declare.
Second Cancer Incidence Among Chronic Lymphocytic Leukemia (CLL) Patients: A French Population-Based Study
Authors: Cornet Edouard , Jégu Jérémie , Mounier Morgane , Maynadié Marc , Monnereau Alain , Velten Michel , Troussard Xavier 
Published: October 2019

Abstract
Introduction. The risk of developing a second primary cancer (SPC) is increased in patients with CLL. The mechanisms explaining this association could be related to lifestyle, environment, host factors or interactions or other influences. We conducted an epidemiological study based on 10 French registries and evaluated the risk of developing SPC in patients with CLL.
Methods. Data from French population-based registries were used to establish a cohort of all patients diagnosed with a first cancer between 1989 and 2004 and followed-up until December 31, 2007. The person-year approach was used to estimate Standardized Incidence Ratios (SIRs) and Excess Absolute Risks (EARs) of metachronous SPC. Multivariate Poisson regression modules were then used to model SIRs and EARs separately by gender, adjusting for age, year of diagnosis, follow-up and first cancer site. All patients with CLL did not have HIV/AIDS-related disease.
Results. Among 288,967 patients, 21,226 patients (7.5%) developed SPC. Among 4,148 CLL patients (Male: 2,336, Female: 1,812, median age: 70 years), 479 patients (11.5%) developed SPC after a median time of 54 months (2-199). EARs of SPC are different between male and female. In male, localizations of SPC are lung, bronchus and trachea, colorectum, colon, stomach and Hodgkin's disease. Their respective EARs are shown in table 1 and no excess absolute risk was observed for 24 other localizations. In female, localizations of SPC are rectum, stomach and melanoma of skin. Their respective EARs are shown in table 2 and no excess absolute risk was observed for 21 other localizations.
Table 1: Excess Absolute Risks (EARs) of second primary cancer (SPC) for male Male SIR SIR IC95% EAR EAR IC95% Lung, bronchus and trachea 2.08 1.61 2.64 26.1 14.8 39.8 Colorectum 1.54 1.15 2.03 13.2 3.7 25.1 Colon 1.63 1.11 2.3 9.3 1.6 19.2 Stomach 1.91 1.07 3.15 5.4 0.4 12.7 Hodgkin's disease 8.71 2.34 22.29 2.7 0.5 7.4
Table 2: Excess Absolute Risks (EARs) of second primary cancer (SPC) for female Female SIR SIR IC95% EAR EAR IC95% Rectum 2.17 1.19 3.64 6.7 1.1 15.2 Stomach 3.04 1.45 5.58 6 1.3 13.4 Melanoma of skin 2.59 1.04 5.34 3.8 0.1 10.4
Conclusion. The risk for developing SPC after CLL is increased for male and female, especially solid cancers. Except Hodgkin's disease for male patients, there is no increase for other malignant hematological diseases, including acute leukemia and myelodysplastic syndrome. CLL survivors face a high risk of new malignancies and the excess risk of SPC increases with follow-up. As the SPC risk is closely tied to patient's characteristics, a personalized surveillance is required to allow optimal SPC prevention and early detection strategies.

Disclosures
No relevant conflicts of interest to declare.
Combined Inhibition of Wee1 and Poly(ADP Ribose) Polymerase1/2 Synergistically Inhibits Acute Myeloid Leukemia Cells By Enhancing DNA Damage and the Induction of Apoptosis
Authors: Snedeker Jonathan C , Burleson Tamara M , Tibes Raoul , Porter Christopher C. 
Published: October 2019

Abstract
Introduction: Successful treatment of AML remains dependent upon cytotoxic chemotherapy. However, traditional regimens are not well tolerated by older patients who are at highest risk of disease, and salvage rates after relapse are low, necessitating novel therapeutic strategies. Our groups identified Wee1 as a potential therapeutic target in AML, particularly in the context of concomitant treatment with cytarabine (Tibes et al, Blood, 2012; Porter et al, Leukemia, 2012). Wee1 inhibits CDK1&2 via phosphorylation thereby stalling cell cycle progression. One consequence of Wee1 inhibition/CDK1 activation is impairment of DNA repair via homologous recombination (Krajewska et al, Oncogene, 2013). Cells in which HR is impaired are dependent upon Parp1/2 function, and HR deficient cells are particularly sensitive to Parp1/2 inhibition. Therefore, we hypothesized that combined Wee1 and Parp1/2 inhibition may result in greater inhibition of AML cell proliferation and survival than either alone.
Methods: Human AML cell lines, MV4-11 and Molm-13, and a mouse AML that expresses MLL-ENL/FLT3-ITD were cultured with various concentrations of a Wee1 inhibitor (AZ1775) and a Parp1/2 inhibitor (olaparib) and counted 72 hours later by propidium iodide exclusion and flow cytometry. In some experiments, cells were split into fresh media to recover for 72 more hours. Combination Index (CI) values were calculated by the method of Chou and Talalay. Apoptosis was measured using Annexin V/7AAD and flow cytometry. Western blots were used to confirm inhibition of CDK1/2 phosphorylation and to measure DNA damage induction (gamma-H2AX).
Results: Combined inhibition of Wee1 and Parp1/2 was synergistic, as measured by cell numbers at 72 hours, in all 3 cell lines tested, with combination index values ranging from 0.3 to 0.9. When cells were allowed to recover after treatment, those treated by single agents were able to continue proliferating. However, those treated with the combination did not recover as well or at all, indicating greatly impaired proliferative capacity. Combined inhibition of Wee1 and Parp1/2 also resulted in a significant increase in apoptosis greater than either drug alone. Western blots for gamma-H2AX confirmed that the combination of Wee1 and Parp1/2 resulted in more DNA damage than either drug alone.
Discussion: Combined inhibition of Wee1 and Parp1/2 results in greater inhibition of AML cell proliferation, DNA damage and apoptosis than either drug alone. Future studies will include experiments with primary patient samples, as well as in vivo trials combining Wee1 inhibition with Parp1/2 inhibition. These preliminary studies raise the possibility of rational combinations of targeted agents for leukemia in those for whom conventional chemotherapeutics may not be well tolerated.

Disclosures
No relevant conflicts of interest to declare.
The F-BAR Protein PACSIN2 Regulates Platelet Intracellular Membrane Architecture and in Vivo Hemostatic Functions
Authors: Jönsson Terese , Pluthero Fred G , Jurak Begonja Antonija , Kormann Jan , Demers Mélanie , Wagner Denisa D , Plomann Markus , Hartwig John H , Kahr Walter H , Falet Hervé 
Published: October 2019

Abstract
Proteins of the Bin-amphiphysin-Rvs (BAR) and Fes-CIP4 homology BAR (F-BAR) families bind and deform membranes, promoting in cells tubular invaginations reminiscent of the platelet open canalicular system (OCS) and megakaryocyte (MK) demarcation membrane system (DMS). Here we investigated the role of the F-BAR protein PACSIN2 in platelets and MKs. Spinning disk laser fluorescence confocal microscopy showed that PACSIN2 co-localized with the von Willebrand factor receptor subunit GPIbα at the openings of the OCS in platelets. Endogenous and over-expressed PACSIN2 associated with anastomosing tubular structures reminiscent of the pre-DMS in mouse bone marrow MKs. Pacsin2-null mice had mild thrombocytopenia and polycythemia, MK hyperplasia and splenomegaly. Pacsin2-null mice had a bleeding disorder, as evidenced by significantly prolonged tail bleeding and delayed in vivo thrombus formation following FeCl3 vascular injury, which was normalized by injection of WT platelets. However, Pacsin2-null platelets expressed P-selectin and activated the integrin αIIbβ3 normally in response to stimulation with thrombin and the GPVI-specific collagen-related peptide. In contrast, Pacsin2-null platelets had abnormal flattened morphology, with more and narrower OCS channels on their surface, compared to WT platelets. Together, the data indicate that the F-BAR protein PACSIN2 orchestrates the intracellular membrane architecture of platelets, thereby regulating their in vivo hemostatic functions.

Disclosures
No relevant conflicts of interest to declare.
TMEM16F Deficiency Causes a Strain-Dependent Lethality in Mice and Reduced Phosphatidylserine Exposure on Heterozygous As Well As Homozygous Deficient Mouse Platelets
Authors: Kretz Colin A , Novakovic Valerie A , Kiseleva Anna , Cleuren Audrey C.A. , Khoriaty Rami , Adams Elizabeth , Ginsburg David , Gilbert Gary E. 
Published: October 2019

Abstract
Scott Syndrome is a rare bleeding disorder characterized by a defect in platelet phosphatidylserine (PS) exposure. The syndrome has recently been linked to mutations in TMEM16F, a Ca++-activated ion channel. Tmem16f-/- mice were recently reported to be viable with a prolonged tail snip bleeding time but no spontaneous bleeding (Yang et al, Cell 151: 111-122; 2012). We now report analysis of an additional gene targeted Tmem16f allele generated in C57BL/6 ES cells.
JM8 ES cell were obtained from EUCOMM, and successful Tmem16f gene targeting in intron 1 was confirmed by PCR and sequencing. Genotyping of 120 Tmem16f+/gt (+/gt) intercross progeny identified no surviving Tmem16fgt/gt (gt/gt) mice at weaning (p<0.001). However, +/gt intercrosses generated the expected Mendelian genotype ratios at both E10.5 and E17.5, with gt/gt embryo’s exhibiting no morphological abnormalities on gross or routine histologic examination. Though complete deficiency of TMEM16F is lethal in the C57BL/6J genetic background between E17.5 and birth, an F2 intercross of +/gt mice outcrossed one generation to 129x1SvJ resulted in gt/gt mice surviving to weaning, though at reduced numbers (6/75 total progeny compared to ~19 expected, p <0.002). Progeny testing of surviving gt/gt mice suggest a single autosomal dominant 129x1SvJ-associated genetic modifier. Preliminary genetic analysis of these mice appears to map this locus to the proximal region of chromosome 3.
Tail bleeding times for gt/gt were >10min, whereas littermate +/gt and +/+ mice bleeding ceased at 8 ± 1 min and 6 ± 0.8 min, respectively, each significantly different than gt/gt (p<0.05). Notably, platelets from +/gt mice exhibited a trend toward reduced PS exposure, detected with FITC-labelled lactadherin, in response to PAR4 agonist peptide, whereas gt/gt mice had significantly reduced PS exposure (p < 0.05). gt/gt platelets showed a trend toward reduced PS exposure in response to A23187, as well as prolonged platelet rich plasma clotting times, and less efficient lactadherin inhibition of platelet clotting time.
These data suggest the existence of a viability-determining genetic modifier of TMEM16F in the 129x1SvJ mouse strain. Identification of the responsible gene may uncover novel functions for TMEM16F and the regulation of hemostatic function. The observation of a PS exposure and hemostatic phenotype in +/gt mice also suggests the possibility that heterozygous TMEM16F mutations may subtly influence hemostasis or thrombosis in humans.

Disclosures
No relevant conflicts of interest to declare.
Platelet-Derived Proteases ADAM10 and ADAM17 Impair NK Cell Immunosurveillance of Metastasizing Tumor Cells By Diminishing NKG2D Ligand Surface Expression
Authors: Raab Stefanie , Kropp Korbinian Nepomuk , Steinle Alexander , Klein Gerd , Kanz Lothar , Kopp Hans-Georg , Salih Helmut R 
Published: October 2019

Abstract
Introduction: NK cells are cytotoxic lymphocytes the reactivity of which is governed by the principles of ‘missing-self’ and “induced-self’ recognition. This implies that cells with low or absent expression of MHC class I and/or stress-induced expression of ligands for activating receptors like NKG2D (NKG2DL) are preferentially eliminated by NK cells. We and others reported previously that NKG2DL undergo proteolytic cleavage (i.e. shedding) which promotes evasion from NKG2D-mediated tumor immunosurveillance. Notably, the reactivity of NK cells is also influenced by various other components of the hematopoietic system, and we and others provided evidence for the involvement of platelets in tumor immune evasion including impairment of NKG2D-mediated immune surveillance (e.g., Kopp et al., Cancer Res 2009, Raab et al., Blood 2013 122:3488). Here we extend our recent findings and provide further data how platelets affect immunostimulatory NKG2D-NKG2DL interaction. Methods: Tumor cells were incubated with platelets from healthy donors or treated with platelet-derived soluble factors (releasate) obtained by stimulation of platelets with known platelet agonists including ADP and thrombin. NKG2DL and ADAM10 as well as ADAM17 surface expression on tumor cells and platelets, respectively, was measured by FACS, while release of NKG2DL was determined by ELISA. ADAM10 and 17 protein levels were assessed by immunoblotting. NK cell lysis of tumor cells in the presence or absence of coating platelets or platelet releasate was determined by chromium release assays. Results: We found that interaction of platelets with tumor cells resulted in substantially reduced NKG2DL expression on the surface of the malignant cells, which was paralleled by enhanced release of soluble NKG2DL. Similar albeit weaker effects were observed upon treatment of tumor cells with platelet releasate, indicating that platelet-derived factors mediate NKG2DL shedding from the tumor cell surface. Notably, ADAM10 and ADAM17, the known sheddases of NKG2DL, were found to be expressed on the platelet surface, and sheddases could also be detected in platelet releasate, pointing to an involvement in platelet-mediated NKG2DL shedding. Diminished NKG2DL surface expression resulted in reduced NKG2D-mediated NK cell cytotoxicity as revealed by blocking experiments using NKG2D antibody and F(ab)2 fragments specific for the modulated NKG2DL. Conclusion: We propose that induction of NKG2DL shedding constitutes a novel mechanism by which the interaction of platelets with metastasizing tumor cells impairs NK cell immunosurveillance.

Disclosures
No relevant conflicts of interest to declare.
uPAR Directly Interacts with LDLR-like Proteins to Induce Angiogenic Endothelial Cell Behavior
Authors: Pausz Clemens , Mawas Rula , Unseld Matthias , Chilla Anastasia , Novotny René , Prager Gerald W 
Published: October 2019

Abstract
In this study we characterized a conserved motif of domain 3 of the urokinase-type plasminogen activator receptor (uPAR) to directly interact with low-density lipoprotein receptor (LDLR)-related protein (LRP) family proteins, thereby affecting endothelial cell motility and angiogenesis in vitro and in vivo.
There is increasing evidence that uPAR plays a central role in growth factor induced endothelial cell activation. Beside its proteolytic role, urokinase-type plasminogen activator (uPA) / uPAR-complex formation induces intracellular signal transduction, which leads to endothelial cell migration and invasion. Since uPAR is a GPI-anchored protein, an interaction with transmembrane proteins - such as members of the LDL-receptor family - is required, inducing signal transduction but also regulating distribution of uPAR via its internalization and recycling to the leading edge. Recently, a direct interaction between uPAR and LRP-family members has been suggested to be sufficient to mediate internalization of uPAR-complex. A crystal structure analysis revealed a small sequence of domain 3 (D3) of uPAR, to be highly exposed upon uPA binding to its receptor.
Applying affinity chromatography analysis as well as mutation expression studies, we identified the sequence as an LRP-binding motif, which affects endothelial cell spreading, migration and invasion upon VEGF in vivo as well as in vitro.
In detail, matrigel-filled angioreactors with embedded retroviral constructs, carrying wild-type or modified uPAR genes, were implanted subcutaneously into uPAR deficient C57BL/6 mice. After explantation, blood vessel in-growth analysis revealed that only angioreactors with reconstituted wild-type uPAR but not reactors with modified uPAR, being deficient in LDLR interaction, showed angiogenesis.
To test a therapeutic impact, peptides mimicking the binding motif and competitive for LDLR binding were used. We found that in a dose dependent manner the peptides did not only block uPAR/LDLR-like protein interaction, but were also capable of blocking VEGF-induced endothelial cell migration in vitro.
In summary, our data show that a conserved motif of uPAR domain 3 is capable to interact with LDLR-like proteins, which is required for efficient growth-factor induced endothelial cell behavior. Preliminary functional data suggest that this extracellular motif might be a potential therapeutic target in angiogenesis dependent diseases such as cancer.

Disclosures
No relevant conflicts of interest to declare.
Bradykinin Receptors Regulate Proliferation, Migration and Tube Formation of Endothelial Progenitor Cells
Authors: Wu Yi , Backdash Nadia , Colman Robert W. 
Published: October 2019

Abstract
Bradykinin is a potent activator of endothelial cells and is known to induce vasodilation, vascular permeability, and nitric oxide production. Bradykinin signaling is initiated by binding to G protein-coupled bradykinin receptors 1 and 2 (B1R and B2R). In this study we investigated whether these two receptors are involved in regulation of endothelial progenitor cell (EPC) biology. First we isolated CD34+ cells from wild-type (WT) and double B1R and B2R knockout (B1RB2R-/-) mice, and compared their capacities of proliferation, migration, as well as tube formation in a 3D cell culture system. Bone marrow-derived B1RB2R-/- EPCs displayed a reduction in proliferation and migration through collagen-coated transwell, compared with EPCs from WT mice. Moreover, implanted B1RB2R-/- EPCs into immunocompromised NOD-SCID mice showed significantly less vasculogenesis in matrigel plug. Human EPCs were isolated from peripheral blood of healthy donors using CD34+ selection. In a concentration-dependent manner, bardykinin stimulated proliferation, migration and tube formation of human EPCs. In contrast to human EPCs, human umbilicial vein endothelial cells (HUVECs) did not display a significant increase in proliferation, migration, and tube formation upon bradykinin stimulation, suggesting that bradykinin selectively enhances the function of EPCs, but not mature endothelial cells. Pre-incubation with a B2R selective antagonist (Icatibant) reduced these functions of human EPCs. When human EPCs were stimulated with [des-Arg9]-BK, a selective B1R agonist, their proliferation and tube formation remained unchanged. Moreover, preincubation of human EPCs with [des-Arg10]-Icatibant, a B1R selective antagonist, did not affect their proliferation, migration and tube formation. As detected by Western blot and real time RT-PCR, B2R was constitutively expressed in EPCs, while the expression of B1R was only induced when stimulated by inflammatory stimuli, such as TNFα and IL-1β. Take together, bradykinin regulates proliferation, migration and tube formation of EPCs through constitutively expressed B2R, whereas B1R is probably involved in inflammatory settings.

Disclosures
No relevant conflicts of interest to declare.