bluuebunny/crossref_metadata_2025_split huggingface dataset:
PS1453 CURCUMIN INHIBITS PROLIFERATION AND INDUCES APOPTOSIS IN JAK2 MUTATED CELLS THROUGH THE INHIBITION OF JAK2/STAT AND MTORC1 PATHWAYS
Authors: Petiti J. , Rosso V. , Iacono M. Lo , Panuzzo C. , Fava C. , Cilloni D. 
Published: June 2019

Background:Myeloproliferative neoplasms are chronic myeloid cancers divided in Philadelphia positive and negative. The JAK2 V617F is the most common mutation in Philadelphia negative patients and results in a constitutive activation of the JAK/STAT pathway, conferring a proliferative advantage and apoptosis inhibition. Furthermore, recent studies identified a functional crosstalk between the JAK/STAT and mTOR pathways. Given the heterogeneous clinical needs of MPNs patients, identification of an effective therapy is often difficult. Therefore, the availability of new therapeutic approaches might be very attractive. Curcumin is the active phytochemical component of the Curcuma longa plant. Previous studies showed that curcumin can suppress JAK2/STAT signaling pathways in different type of cancer and injuries.Aims:In this study, we investigated the anti‐proliferative and pro‐apoptotic effects of curcumin in JAK2 V617F cell line and in primary cells.Methods:HEL cell line and cells from patients affected by myeloproliferative neoplasms have been incubated with increasing concentrations of curcumin for different time. Apoptosis and proliferation were evaluated. Subsequently, JAK2/STAT and AKT/mTOR pathways were investigated at both RNA and protein levels.Results:We showed that curcumin strongly inhibited the proliferation and induced apoptosis in a dose and time dependent manner in HEL cells. Furthermore, we found that curcumin markedly reduced JAK2 phosphorylation and, consequently, the activation of its downstream effectors STAT3 and STAT5 in HEL cells. We observed that JAK2/STAT inhibition induced the down regulation of Pim family members in HEL cell line and in patient specimens. Farther, our results in HEL cells showed that curcumin negatively regulated the mTORC1 complex formation. As a consequence, we observed a strong inhibition of mTORC1 downstream effectors 4eBP1 and p70s6K phosphorylation. Our preliminary data on cells from MPNs patients showed that the in vitro curcumin treatment significantly promoted apoptosis and reduced the expression of Pim family members, bringing back their mRNA expressions to comparable levels with healthy subjects ones. Furthermore, the curcumin effect was strengthened by the observation that CD177, an antigen over expressed in neutrophils of the majority of MPNs patients, was down regulated by curcumin.Summary/Conclusion:In conclusion, the present study showed that curcumin exerts an antitumor effect on human JAK2 mutated cells by inducing apoptosis and inhibition of proliferation, through the regulation of both JAK2/STAT and mTORC1 pathways. These findings suggest that curcumin seems to be a promising nutraceutical compound that should be further evaluated in different pharmaceutical formulation for the treatment of MPNs.
PS1469 IL28B GENE POLYMORPHISMS INFLUENCE ON THE RESPONSIVENESS AND TOLERABILITY OF IFN‐Α IN PATIENTS WITH MPN
Authors: Olkhovskiy∗ I. , Stolyar M. , Gorbenko A. , Lagutinskaya D. , Dunaeva E. , Mironov K. , Shipulin G. , Olkhovik T. , Mikhalev M. , Vasiliev E. , Smelyanskaya M. , Hazieva A. 
Published: June 2019

Background:Interferon‐α (IFN‐α) provides several benefits to myeloproliferative neoplasms (MPNs) patients and leads 35% of its experienced a good molecular response. Both newly diagnosed and previously treated patients with polycythemia vera (PV) exhibit high hematologic response rates, and some of these patients achieve molecular responses [Kiladjian J‐J., et al., 2016]. Nevertheless, IFN‐α therapy is often accompanied by serious side effects, which forces some patients to refuse treatment. Only one scientific team [Lindgren M., et al., 2012, 2018] previously published data demonstrating the effect of IL28B gene polymorphisms on the success of interferon therapy in MPNs.Aims:Evaluate the prognostic impact of IL28B rs12979860 and rs8099917 on IFN‐α treatment response and toxicity in patients with MPNs.Methods:35 patients were investigated: 18 with polycythemia vera, 12 with essential thrombocythemia and 5 with primary myelofibrosis. All patients were treated with IFN‐α (three times weekly). DNA from whole blood samples was isolated using «DNAsorb‐B» (Central Research Institute for Epidemiology, Russia). The rs12979860 and rs8099917 SNPs genotyping was determined by pyrosequencing on a PyroMark Q24 instrument. Allele burden JAK2 V617F was determined using «Myeloscreen JAK2» («Formula of gene Ltd», Russia). Hematologic response was evaluated using the European Leukemia Net criteria. The quantitative values are given as a median and 25% and 75% percentiles.Results:The JAK2 V617F allele burden after 60 months of IFN‐α therapy was significantly reduced in samples of patients with polycythemia vera in the presence of a “strong” TT/CC variant of the haplotype IL28B (13; 8.4–24.4) compared with other variants of polymorphism of these gene (57; 44,3–67,0). The average duration of therapy to failure due to side effects of IFN‐α was also 2–3 times shorter (p < 0.05) in patients with a TT/CC haplotype. The therapeutic impact of IFN‐α in patients with MPN involve IL28B signaling pathways. This is the second independent confirmation of the data by Lindgren et al.Summary/Conclusion:Genotype evaluation of IL28B can be useful for predicting the effectiveness of interferon therapy in MPNs.image
PS1477 PLATELET DISFUNCTION IN NOONAN AND 22Q11.2 DELETION SYNDROMES IN CHILDHOOD
Authors: RUIZ A. , ISOLA I. , GASSIOT S. , CATALÀ A. , DÍAZ‐RICART M. , MARTÍNEZ‐MONSENY A.F. , SERRANO M. , BERRUECO R. 
Published: June 2019

Background:An underlying thrombocytopathy seems to be the responsible for hemorrhagic symptoms in patients diagnosed with 22q11.2 deletion syndrome (22q11DS) and Noonan syndromes (NS). Whereas in 22q11DS it is explained by a defect in the membrane glycoprotein complex Ib‐V‐IX, the cause of thrombocytopathy in NS remains unclear.Aims:To study the incidence of thrombocytopathy in pediatric patients diagnosed with 22q11DS or NS assessing the utility of ISTH‐BAT questionnaire and laboratory techniques in this regard.Methods:Prospective study between March and December 2018 in children (2 to 18 year‐old) diagnosed with 22q11DS or NS. Hemorrhagic symptoms, total cell blood count, platelet indices, closure time and platelet aggregation were evaluated in all patients. Membrane glycoprotein expression was only performed in 22q11DS patients.Results:Among NS patients (n = 22), near 80% had an aggregation defect with adenosine diphosphate (ADP) and epinephrine without thrombocytopenia. A significant correlation (p < 0.05) between closure times and aggregation with arachidonic acid, epinephrine and ADP was found. This correlation was more evident in those patients with hemorrhagic symptoms.Five out 29 (17%) patients diagnosed with 22q11DS had thrombocytopenia. Impairment in ristocetin‐induced platelet aggregation was found. This defect was correlated with prolonged closure times. Moreover, a significant correlation between thrombocytopenia, closure times and hemorrhagic symptoms was demonstrated.Summary/Conclusion:This study shows in a cohort of patients with NS and 22q11DS a significant impairment of platelet aggregation which correlates with hemorrhagic symptoms and closure times. Thus, closures times might be useful as screening test to evaluate the bleeding risk in this group of patients.
PS1496 ANKRD26‐RELATED THROMBOCYTOPENIA: STUDY OF 3 FAMILIES
Authors: Coelho D. Pereira , Azevedo J. , Martinho P. , Nascimento T. , Marini S. , Ribeiro A. Barbosa , Cortesão E. , Luís D. , Rito L. , Sevivas T. , Guilherme R. , Fidalgo T. , Ribeiro L. 
Published: June 2019

Background:ANKRD26‐related thrombocytopenia (ANKRD26‐RT) is a non‐syndromic form of inherited thrombocytopenia with autosomal dominant transmission, caused by mutations in the 5’UTR region of the ANKRD26 gene. About 8–10% of the patients develop myeloid neoplasms. Recently, in the 2016 revision of the WHO, this entity was recognized in the group of myeloid neoplasms with germ line predisposition and preexisting platelet disordersAims:Characterization of 3 portuguese families with ANKRD26‐RT.Methods:Study of 3 families with thrombocytopenia and ANKRD26 gene mutation. Initial study on NGS platform (Ion Torrent ™, Thermo Fisher Scientific), subsequent confirmation/family studies by Sanger direct sequencing. Clinical (symptoms, treatment, evolution) and laboratory data (platelets, MPV, Hb, leukocytes, bone marrow) were analyzed for 11 individuals.Results:Mutations in the 5’UTR region of the ANKRD26 gene in heterozygosity were found in 11 individuals studied, belonging to 3 families with autosomal dominal familial thrombocytopenia and normal MPV.: c.‐118C>T mutation in family 1 and 2, and the more recently described c.‐140C>G mutation in family 3. Median age at diagnosis of thrombocytopenia was 16 years (0.25–55), with a current median age of 36 years (2–73). In those affected, the median number of platelets is 48x109L (8–203), with persistently normal MPV in 82% of patients (8,2–12,9). In our cohort, the elderly (> 65 years) had lower platelet counts (<15x109L) and 1 family member with the c.‐140C>G mutation had normal platelet count in one determination. About half of the patients had Htc> 45% (max 50.4%). More than half of the patients (54.5%) were initially interpreted as ITP and treated with prednisolone or IV Ig, with no response. Only 5 patients experienced bleeding events, two requiring platelet transfusion. The patient with the most significant hemorrhagic symptoms (45 years old, platelets 15–30x109L) was classified as MDS in 2011.Summary/Conclusion:ANKRD26‐RT is associated with a risk of developing myeloid neoplasia, making its diagnosis and monitoring particularly important. The recognition of this entity is recent and probably underdiagnosed. In our center we identified 3 ANKRD26‐RT families, 2 of them had 3 affected generations. Its characterization and follow‐up will allow us to better understand the variability of presentation and evolution and possibly the identification of bio‐clinical risk factors for neoplastic transformation.
PS1515 PREMEDICATION WITH MONTELUKAST AND RUPATADINE DECREASES INFUSION TIME, RATE AND SEVERITY OF REACTIONS AND OVERALL COST OF RITUXIMAB ADMINISTRATION
Authors: Kotchetkov R. , McLean J. , Gerard L. , Nay D. , Hopkins S. 
Published: June 2019

Background:Rituximab is associated with frequent infusion reactions, which carry significant burden to both patients and health care practitioners. Standard pre‐medications (SP) do not prevent reactions sufficiently. Montelukast (M) and Rupatadine (R) are used for treatment of urticarial, allergic rhinitis, and may impact Rituximab infusionAims:We assessed impact of addition of R, M and their combination (R+M) on Rituximab infusion/rate, rate/severity of reactions and administration costMethods:Adult patients with lymphoproliferative disorders treated at our cancer center between Jan 2018 to Jan 2019 were evaluated with Rituximab‐containing regiments. Since the majority of reactions occur during the first infusion, our study was limited to the initial Rituximab treatment. Patients received either SP with diphenhydramine/acetaminophen and additional R, M or R+M combination. Comparative analysis of infusion time/rate, severity of infusion reactions, number of rescue medications and cost of Rituximab infusions among groups was performed using one‐way ANOVA with Tukey post‐hoc or chi‐square. The study was approved by our institutional ethics boardResults:Patients received either: 1) standard premedications (SP); 2) SP + Rupatadine (R) 10 mg; 3) SP + Montelukast (M) 10 mg; or 4) both (SP+R+M). Patient characteristics and therapy are shown in Table 1. Compared to SP, the R, M and R+M groups had greater improvement in Rituximab delivery. Mean infusion time was 305 [range 235–441] min. in SP, 254 [105–390] in M, 265 [193–350] in R and 238 [196–342] in R+M groups, (p = 0.0001). Infusion reactions occurred in 92% in SP vs. 38, 45, 31% in M, R and R+M groups (p = 0.0001). Median reaction grade was 2 in SP, 1 (M), 0 (R and R+M). Median number of rescue medications was 3 [0–10] in SP vs 0 [0–7] in M, R and R+M groups. Cost of rescue medications (US$) was 41 [0–63] (SP), 11 [0–50] (M), 17 [0–63] (R), 7 [0–58] (R+M) groups (p < 0.0001). Mean nursing cost (US$) per patient infusion was calculated as 269 [207–388] in SP vs 222 [92–343] (M), 233 [170–308] (R), 209 [174–301] (R+M) group.RCHOP, Rituximab/Cyclophosphamide/Doxorubicin/ Oncovin/Prednisone; BR, Bendamustine/Rituximab; FCR, Fludarabine/Cyclophosphamide/RituximabimageSummary/Conclusion:Addition of R, M and particularly R+M combination significantly improved Rituximab delivery, decreased the rate and severity of infusion reactions, and lowered the cost of Rituximab administration
PS1522 CENTRAL PHYSIOLOGIC MECHANISMS WHICH AUGMENT OXYGEN RELEASE (BOHR EFFECT AND 2,3‐DPG BINDING) ARE PRESERVED IN THE PRESENCE OF VOXELOTOR AT THE THERAPEUTIC TARGET OF 30% HB MODIFICATION
Authors: Pochron M. , Siu V. , Oksenberg D. , Dufu K. 
Published: June 2019

Background:Key physiologic mechanisms that drive O2 offloading to tissues in vivo include the Bohr effect and 2,3‐DPG binding. The Bohr effect is the capacity of Hb to offload more O2 in hypoxic peripheral tissues in response to conditions of increased H+ concentrations (or acidosis). Similarly, the binding of 2,3‐DPG to Hb stabilizes the deoxyHb state, reduces the Hb‐O2 affinity and facilitates O2 release to tissues.Aims:To investigate oxygen off‐loading dynamics from voxelotor‐modified hemoglobin in vitro under increased acidity or increased DPG content.Methods:In this study, the Oxygen Dissociation Assay (ODA) and oxygen equilibrium curves (OECs) were used to determine the effect of voxelotor on Hb‐O2 affinity in response to H+ and 2,3‐DPG (5 mM). The ODA is a spectrophotometric assay that measures the time dependences of O2 release, while Hb‐O2 affinities can be measured using OECs. Studies in the ODA were conducted with purified Hb (3 μM), while OEC determinations were conducted with both purified Hb (25 μM) and whole blood.Results:Relative to Hb alone, voxelotor‐modified Hb dose‐dependently increased the proportion of oxygenated Hb (oxyHb) molecules during two hours of deoxygenation consistent with stabilization of hemoglobin in the oxy, or R‐ state. Under deoxygenated conditions, the percentage of oxygenated voxelotor‐modified Hb decreased over time, demonstrating oxygen release (offloading) by voxelotor‐modified Hb. With increased acidity (down to 6.8), there was an acceleration of oxygen release from voxelotor‐modified Hb under deoxygenated conditions. This resulted in an increase in the p50 of voxelotor‐modified blood and thus demonstrates a pH‐dependent decrease in the affinity of Hb for O2 in the presence of voxelotor at the therapeutic target Hb modification of 30% (Figure 1A). Similarly, the response of Hb to 2,3‐DPG effect was not inhibited in the presence of voxelotor at the 30% modification target (Figure 1B).Summary/Conclusion:The results of this study indicate that the central physiologic mechanisms which augment oxygen release to metabolically active tissues, such as the Bohr effect and 2,3‐DPG binding, are preserved when voxelotor is bound to Hb.image
PS1537 CO‐TRANSPLANTATION OF MESENCHYMAL STEM CELLS CAN AMELIORATES ACUTE GVHD AND VIREMIA AFTER ALLO‐HSCT FOR APLASTIC ANEMIA: A MULTI‐CENTER RETROSPECTIVE STUDY OF 75PATIENTS
Authors: Chen M. , Zheng Z. , Hu J. , Yang T. 
Published: June 2019

Background:Mesenchymal stem cells (MSCs) are known to exhibit immunomodulatory, anti‐inflammatory, and pro‐angiogenic properties, and therefore have the potential to improve the outcome of allogeneic hematopoietic stem cell transplantation (allo‐HSCT) for severe aplastic anemia (AA).Aims:To explore the efficacy and safety of allo‐HSCT by co‐transplantation with MSCs in AA patientsMethods:We conducted a multi‐center retrospective study to comparing the incidence and severity of acute GvHD, transplant‐related complication and overall survival (OS) of allo‐HSCT with or without co‐injection of MSCs in AA patients. A total of 75 consecutive severe AA patients (43males/32 females) undergoing allo‐HSCT at 4 hospitals from January 2012 to February 2018 were analyzed. 45 patients transplanted with donor HSCs only,and 30 patients transplanted with HSCs and MSCs. MSCs infusion dose was 0.5–3.0 × 106/kg, at day0/+1 following HSCs infusion.Results:All patients achieved successful engraftment within one month post transplant. There was no difference in time to leukocyte and platelet engraftment in the two groups. There was a trend to a lower incidence of aGVHD in the MSC group (13.3% versus 26.7%,χ2 = 2.066 P = 0.151). Moreover, a lower incidence of CMV replication in the MSC group was observed (CMV:26.7% vs 64.4 %,P = 0.003). With a median follow‐up time of 615 days (range:29–2315ays),the 2‐yr OS was similar in both groups (76.2% vs 77.3%,P = 0.883). A combination of bone marrow and peripheral blood as the sources of stem cells co‐transplanted with MSCs (n = 9) demonstrated an improved survival benefit (2‐yr OS = 100%).Summary/Conclusion:These data support that co‐transplantation with MSCs could ameliorate the challenges of aGVHD and viremia and facilitate survival in allo‐HSCT for AA patients.
PS1567 UNCOUPLING INK PROTEINS FROM CDK4/CDK6 IN HSCS – UNCOUPLING PROLIFERATION FROM SENESCENCE
Authors: Prchal‐Murphy M. , Bellutti F. , Sexl V. , Kollmann K. 
Published: June 2019

Background:Cyclin‐dependent kinase (CDK) 4 and CDK6 are highly related cell‐cycle kinases that regulate progression from G1 to S‐phase. CDK4/6 are activated upon binding of D‐type cyclins and subsequently phosphorylate the retinoblastoma protein (Rb). CDK4/6 inhibitor proteins, the INK4 family (p16INK4a, p15INK4b, p18INK4c, and p19INK4d) as major player, adds another layer of complexity. Hyperactivity of CDK4 or CDK6 is frequently observed in hematopoietic malignancies due to loss of p16INK4a. Besides, CDK6 has also been identified as transcriptional regulator driving complex transcriptional programs. CDK4/6 inhibitors are approved in breast cancer treatment and are currently tested in hematopoietic malignancies.Aims:We aimed to unravel the consequences of the loss of these inhibitory INK‐dependent mechanisms for hematopoiesis.Methods:We used knock‐in mice expressing a CDK6 (CDK6R31C, Cdk6

R/R
) and/or a CDK4 (CDK4R24C, Cdk4

R/R
) mutant insensitive to INK4‐mediated inhibition and analyzed stem cell populations. Further we tested the self‐renewal capacity of mutant hematopoietic progenitors under conditions of hematopoietic stress.Results:Despite comparable bone marrow cellularity, we found a significant reduction in LSK cells and HSC populations in mice harboring Cdk4

R/R
or Cdk6

R/R
mutation. The combination of Cdk4

R/R
/Cdk6

R/R
in double‐knock‐in mice rescued these defects in all progenitor stages. Serially transplanted Cdk4

R/R
/Cdk6

R/R
double‐knock‐in HSCs display enhanced stem cell proliferation. They maintain the ability to give rise to both myeloid and lymphoid lineages. In response to hematopoietic stress (5‐FU, serial and competitive total bone marrow transplantation), CDK4R24C/CDK6R31C prevents stem cell exhaustion and show robust recovery in stem cell numbers. Low‐input RNAseq of long‐term stress‐triggered HSCs will unravel mechanisms uncoupling proliferation from senescence.Summary/Conclusion:HSCs with disrupted CDK4/6 – INK4 binding show enhanced proliferation, reduced exhaustion and improved transplant ability. Analysis of the underlying molecular alterations may open novel therapeutic options for CDK4/6 hyperactive hematopoietic malignancies.
PS1572 HEART DISEASE AND PANCREATIC IRON IN THALASSEMIA MAJOR
Authors: Meloni A. , Pistoia L. , Spadola V. , Murgia M. , Quota A. , Carrai V. , Bulgarelli S. , Mattei R. , Pasin M. , Sardella L. , Restaino G. , Preziosi P. , Positano V. , Pepe A. 
Published: June 2019

Background:Some preliminary data have postulated a correlation between pancreatic iron overload and heart iron and function in thalassemia major (TM) patients.Aims:In the present multicenter study we explored systematically the link between pancreatic iron and heart disease in a large cohort of TM patients.Methods:We considered 880 TM patients (467 M, mean age 37.83 ± 10.05 years) enrolled in the E‐MIOT (Extension‐Myocardial Iron Overload in Thalassemia) project. T2∗ measurements were performed over pancreatic head, body and tail and global value was the mean. Myocardial iron overload (MIO) was quantified using a T2∗ segmental approach. Biventricular function parameters were assessed by cine images. Late gadolinium enhancement (LGE) images were acquired to detect myocardial fibrosis.Results:A significant correlation between pancreatic and cardiac iron was reconfirmed in this more numerous population and a normal pancreas T2∗ showed negative predictive value of 100% for cardiac iron.LGE sequences were acquired in 273 TM patients and 84 (30.77%) of them showed macroscopic myocardial fibrosis. Global pancreas T2∗ values were significantly lower in patients with fibrosis (7.38 ± 6.19 ms vs 11.91 ± 9.79 ms; P < 0.0001).Sixty‐four patients had at least one cardiac complication (arrhythmias, heart failure, pulmonary hyperthension, vascular disease). Patients with cardiac complications showed a significant lower global pancreas T2∗ (7.83 ± 5.61 ms vs 12.76 ± 10.34 ms; P = 0.024). Specifically, global pancreas T2∗ values were significantly lower in patients with heart failure as well as in patients with arrhythmias and all patients with one of these two diseases had a pathologic global pancreas T2∗ value (see Figure).Summary/Conclusion:Pancreatic iron is a strong predictor not only for cardiac iron, but also for cardiac complications supporting a more profound link between pancreatic iron and heart disease in TM.image