BACKGROUND:Oxidative stress is a hallmark of many cancers. The increment in reactive oxygen species (ROS), resulting from an increased mitochondrial respiration, is the major cause of oxidative stress. Cell fate is known to be intricately linked to the amount of ROS produced. The direct generation of ROS is also one of the mechanisms exploited by common anticancer therapies, such as chemotherapy.METHODS:We assessed the role of NFKBIA with various approaches, including in silico analyses, RNA-silencing and xenotransplantation. Western blot analyses, immunohistochemistry and RT-qPCR were used to detect the expression of specific proteins and genes. Immunoprecipitation and pull-down experiments were used to evaluate protein-protein interactions.RESULTS:Here, by using an in silico approach, following the identification of NFKBIA (the gene encoding IκBα) amplification in various cancers, we described an inverse correlation between IκBα, oxidative metabolism, and ROS production in lung cancer. Furthermore, we showed that novel IκBα targeting compounds combined with cisplatin treatment promote an increase in ROS beyond the tolerated threshold, thus causing death by oxytosis.CONCLUSIONS:NFKBIA amplification and IκBα overexpression identify a unique cancer subtype associated with specific expression profile and metabolic signatures. Through p65-NFKB regulation, IκBα overexpression favors metabolic rewiring of cancer cells and distinct susceptibility to cisplatin. Lastly, we have developed a novel approach to disrupt IκBα/p65 interaction, restoring p65-mediated apoptotic responses to cisplatin due to mitochondria deregulation and ROS-production.

Hodgkin lymphoma (HL) is an uncommon B-cell malignant disease. It usually presents with mediastinal and/or laterocervical lymph node localization, while primary extranodal HL is a rare entity giving rise to diagnostic and therapeutic challenges. It rarely presents as just extranodal localization, so its presence within the maxillary sinus without any lymphadenopathy is exceptional. Given the rarity of this localization, there is no standard treatment for maxillary sinus HL. We present a case of a patient with extranodal HL of the right maxillary sinus treated with primary surgery followed by adjuvant sequential chemoradiation therapy.

Background: Acute lymphoblastic leukemia (ALL) is a molecularly heterogeneous disease originating from clonal proliferation of precursor B-lineage cells. In adults, ALL diagnosis is still associated with a dismal prognosis due to the lack of specific targeted therapies. This study was designed to investigate the expression of interleukin-2 receptor alpha chain CD25 in B-ALL and its biological significance, especially following the availability of specific CD25 targeting compounds. Methods:The expression of IL2RA (CD25 gene) was detected by flow cytometry (FC), immunohistochemistry and Western blot analysis, in 25 newly diagnosed ALL patients, both Philadelphia positive (12 patients) and Philadelphia negative (13 patients). Similarly, CD25 expression was assessed in four B-ALL commercially available cell lines. Infection with shRNA specifically directed against CD25 was used to evaluate apoptosis induction and cell cycle arrest in primary B-ALL cells established from two patients. Results:Our data suggest that ALL, and in particular Ph-positive ALL, aberrantly expresses the interleukin-2 receptor alpha chain, CD25. Whereas normal B cells display low amounts of CD25, primary ALL cells and ALL cell lines (over)-express CD25. While the high frequency of CD25 on the surface of many different hematological tumor cells has been established and confirmed in our study, there is little investigation focusing on the significance of CD25 expression. Indeed, CD25 may be present on ALL cells and enable oncogenic signaling pathways. In such respect, we observed that CD25 silencing in primary cells promotes cell cycle arrest and apoptosis induction. While these data support the rational to target CD25, ALL cells did not appear to be in-vitro sensitive to basiliximab, an antibody able to target the Il2RA, but in-vivo investigations are needed to better assess the effects of this therapeutic approach in ALL context. Conclusions:We concluded that CD25 expression is elevated in patients with B-ALL. Our results also demonstrate that CD25 silencing induces cell cycle arrest and apoptosis. The latter result has important implications from a therapeutic point of view. Targeting CD25 receptor with anti-CD25 antibodies or peptide mimetics could be an effective strategy for targeting leukemic cells. Additionally, high CD25 expression could be exploited for the development of CAR-T therapy Disclosures Saglio: Roche:Research Funding;Pfizer:Research Funding;Incyte:Research Funding;Novartis:Research Funding;Ariad:Research Funding;Bristol-Myers Squibb:Research Funding.

Acute myeloid leukemia (AML) is a disease of the elderly where only 6.7% of patients >65 years of age are alive at 5 years from diagnosis [1]. This poor prognosis is due to clinical unfitness of older patients to receive standard chemotherapy, to co-morbidities of older age and to the higher incidence of poor prognosis molecular abnormalities [2]. Hypomethylating agents (HMAs), azacitidine and decitabine, are possible options for elderly patients unable to tolerate a systemic chemotherapy [3]. But in light of this poor prognosis, endpoints other than overall survival (OS) such as adverse infection events, transfusion support and in general an improved quality of life must be taken into account [4]. Hence the importance to stratify among old and unfit patients those who can benefit from therapy with HMAs. Immunophenotyping by multiparametric flow cytometry analysis (MFC) is a well-established tool to characterize leukemic cells at diagnosis [5]. Several studies associated adverse prognosis with intraindividual heterogenic leukemic immunophenotype, including few markers like CD58, CD117 and CD14 on overall survival [6,7]. However, AML MFC is not routinely used for patients’ stratification, to predict the course of the disease or responses to treatments. Here, we assessed blast heterogeneity at diagnosis in a cohort of old patients with de novo/secondary AML by MFC, then we evaluated if this phenotypical heterogeneity predicts treatment responsiveness in patients eligibile to HMAs. We retrospectively analyzed a total of 47 newly diagnosed AML (acute promyelocytic leukemia excluded) who referred to our department (Supplementary data and Supplementary Table 1), with informed consents and internal institutional ethical committee-approved protocol (approval number 201/2014). Patients had an age at diagnosis between 65 and 88 years old. Diagnosis was made by histological examination of bone marrow and blasts count by MFC (Supplementary Data). A subdivision of the population was performed based on the expression variability of the surface markers of blast cells at the diagnosis. As described in Supplementary Data, three groups were identified: (a) phenotypically homogeneous (Homo), blasts share the same MCF profile; (b) phenotypically low heterogeneous (HetLow), two to three distinct clones can be separate by MFC (Supplementary Figure 1); (c) phenotypically high heterogeneous (HetHigh), with more than three clones (Supplementary Figure 2). For each patient sex, age, white blood cells (WBC) count, transfusion support, OS, HMAs (azacytidine, decitabine, guadecitabine) were recorded. Patients were equally distributed for age and sex in the three groups. More than 90% of patients in the Homo, 37.5% in the HetLow and 35.2% HetHigh groups were treated with HMAs. In these last two groups standard chemotherapy or therapy of support were prevalent. Considering the non-uniformity of the treatment among this population we extrapolated data of patients treated with HMAs evaluating if heterogeneity of blast profile can predict the prognosis in this subgroup. Analyzing the whole population, we have observed that OS has a mean time of 15.5months in the Homo group, 9.7months in the HetLow group and 4.1months in the HetHigh group. OS at 3months from diagnosis is 92.31% for patients of the Homo group, 73.33% and 52.94% for those owning to the HetLow and HetHigh group respectively. At 12months from diagnosis all patients of the HetHigh group have died while 20.00% of patients of the HetLow group and 38.46% of the Homo group were still alive. Inverse proportional correlation between mean OS and blast phenotypical heterogeneity at diagnosis reached statistical significance considering the Homo and HetHigh group (p-value .0009), the HetLow and HetHigh group (p-value .0027); statistical significance was not reached (p-value .2348) between the Homo and the HetLow group (Figure 1(A)). In azacytidine-treated group,

Background:inflammatory bowel disease (IBD) is characterized by chronic inflammation associated with an increased tendency to thrombosis and thromboembolic complications. The underlying mechanisms of VTE are not yet fully understood. Several studies reported different expressions of circulating procoagulant factors or fibrinolysis inhibitors in IBD. Others linked podoplanin overexpression with thrombosis, hypercoagulability, and increased risk of VTE. Here, we aimed to identify, among genes able to trigger thrombosis, those aberrantly expressed in IBD samples as compared with matched normal mucosa. Methods:we analyzed the transcriptome of matched normal and inflamed lesions in 168 patients with Adult Crohn Disease (CD) and 245 pediatric IBD using publicly available datasets. We intentionally assessed the expression levels of triggers and inhibitors of the coagulation system (Tissue Factor, TFI), the fibrinolytic system (PLAU, PLAT, PAI-1/2/3, TAFI, PN-1, SERPINF2, A2M, SNX1, SERPINC1) and platelets activation (podoplanin). Analyses were finally performed using additional datasets with 219 ulcerative colitis (UC) adult patients, 198 IBD patients and compared to normal colon transciptome. Results:in all datasets, when compared to matched normal mucosal transcriptome, CD and UC inflamed tissues over-expressed Podoplanin mRNA (p=2.46e-25). TF mRNA was also consistently up-regulated in inflamed mucosal of IBD patients, even if paralleled with up-regulation of TFPI as well, questioning on the functional role of TF in IBD. Among the regulators of the fibrinolytic system the urokinase-activator of the plasminogen is consistently deregulated in inflamed areas. Conclusion: our in-silico analyses on gene expression profiles, using matched normal and pathological mucosa of the same patients, suggested that IBD inflamed mucosa favors platelets activation due to podoplanin over-expression. These observations require further studies to assess the biological role of podoplanin in IBD patients and opens new insights on how to perform antithrombotic prophylaxis in IBD patients. Disclosures No relevant conflicts of interest to declare.

The reversal of low-molecular-weight heparin (LMWH) and the management of bleeding patients on LMWH remain highly challenging. Even if LMWH is very extensively administered in the prophylaxis and treatment of venous thrombosis, specific antidotes are lacking, and reversal strategies have very weak grade of evidences on clinical effectiveness. We here describe a reversal strategy with protamine and FVIIa in a patient presenting with hemorrhagic shock and cardiocirculatory arrest.

Background: Primary mediastinal large B cell lymphoma (PMLBCL) is a rare subtype of non-Hodgkin lymphoma mostly diagnosed in young women and is currently recognized as a distinct clinical and biological entity. First line therapy with R-CHOP allows to achieve good remission rates even if chemoresistant cases remain highly challenging from the therapeutic standpoint. As such, we aimed to compare the transcriptome of R-CHOP resistant PMBCL patients to those of chemosensitive patients. Methods:We extracted RNA from embedded paraffin samples and then we performed whole RNA sequencing on 7 patients. Four of them were selected as chemoresistant (Group A), while three of them were classified as responder to R-CHOP (Group B). A First bioinformatics analysis selected a panel of 200 genes significantly differentially expressed in chemoresistant (A) samples versus chemosensitive (B). An unbiased analysis based on different ontology of the genes led to easily identify common signatures that may better profile the two groups of patients. Finally, on a bias analysis, genes were divided into categories in order to identify potential new targets and/or mechanisms of chemoresistance. Results:We identified three genes which may be frankly related to chemoresistance in PMLBCLs due to an overexpression in the group A or a suppression of expression in group B. We selected NFKBIA, the gene which encodes for the IκBα protein, mutated in numerous Hodgkin's lymphoma cells, which cause NFkB to be chronically active in the lymphoma tumor cells. For this reason, we imagine that it could have a major role in the modulation of PMLBCLs sensitivity to chemotherapy. EPHB1 was selected for its very strong expression in the poor prognosis group, since it is one of the most expressed genes and for his association with numerous cancers. The kinase STK 33 appeared overexpressed in group A. A more thorough investigation of this gene might lead to new, significant findings. Conclusions: Our in-silico analyses allowed to identify in PMBCL an unique profile that may modulate sensitivity to chemotherapy. Further analyses may address whether this unique phenotype has clinical implications. We may expect to: i) develop specific therapeutic strategies to target NFkB, EPHB1 and STK33 pathways in PMBCL with a chemoresistant behaviour; ii) correlate the expression profile of resistant PMBCL to other transcriptomes, including those of Hodgkin Lymphomas, Diffuse Large B cell Lymphomas, in order to better profile clinical and biological overlapping features. Disclosures Saglio: Ariad:Research Funding;Incyte:Research Funding;Roche:Research Funding;Bristol-Myers Squibb:Research Funding;Pfizer:Research Funding;Novartis:Research Funding.

Platelet hyperactivation is involved in the established prothrombotic condition of metabolic diseases such as Type 2 Diabetes Mellitus (T2DM) and familial hypercholesterolemia (HC), justifying the therapy with aspirin, a suppressor of thromboxane synthesis through the irreversible inhibition of cyclooxygenase-1 (COX-1), to prevent cardiovascular diseases. However, some patients on aspirin show a higher than expected platelet reactivity due, at least in part, to a pro-oxidant milieu. The aim of this study was to investigate platelet reactivity in T2DM (n = 103) or HC (n = 61) patients (aspirin, 100 mg/day) and its correlation with biomarkers of redox function including the superoxide anion scavenger superoxide dismutase (SOD) and the in vivo marker of oxidative stress urinary 8-iso-prostaglandin F2α. As results, in T2DM and HC subjects the prevalence of high on-aspirin platelet reactivity was comparable when both non-COX-1-dependent and COX-1-dependent assays were performed, and platelet reactivity is associated with a lower SOD activity that in a stepwise linear regression appears as the only predictor of platelet reactivity. To conclude, in T2DM and HC, similarly, the impairment of redox equilibrium associated with a decrease of SOD activity could contribute to a suboptimal response to aspirin.

Background and aims: In the association between hypercholesterolemia (HC) and thrombotic risk platelet hyper-reactivity plays an important role. The inhibition of proprotein convertase subtilisin/kexin type 9 (PCSK9) to reduce plasma LDL-cholesterol merges as effective therapeutic strategy to prevent cardiovascular (CV) events. Aim of this study was to verify whether a treatment up to 12 months with the monoclonal antibodies (mAbs) anti-PCSK9 influences platelet function in primary HC. Methods and results: In patients affected by primary HC (n = 24), all on background of statin and 17 on acetyl salicylic acid (ASA), platelet function parameters were evaluated at baseline up to 12 months of treatment with the mAb anti-PCSK9 alirocumab or evolocumab. From baseline, the treatment with anti-PCSK9 mAbs: i) in ASA HC patients, significantly decreased platelet aggregation detected in platelet-rich plasma by light transmission aggregometry and in whole blood Platelet Function Analyzer-100 assay; ii) in all HC patients, significantly decreased platelet membrane expression of CD62P and plasma levels of the in vivo platelet activation markers soluble CD40 Ligand, Platelet Factor-4, and soluble P-Selectin. Furthermore, CD62P expression, and sP-Selectin, PF-4, sCD40L levels significantly correlated with serum PCSK9. Conclusion: Besides markedly lowering LDL-c levels, our results suggest that HC patients benefit from anti-PCSK9 mAb treatment also for reducing platelet reactivity and increasing platelet sensitivity to the inhibitory effects of aspirin. These effects on platelets could play a role in the reduction of CV event incidence in patients treated with PCSK9 inhibitors. (C) 2019 The Italian Society of Diabetology, the Italian Society for the Study of Atherosclerosis, the Italian Society of Human Nutrition, and the Department of Clinical Medicine and Surgery, Federico II University. Published by Elsevier B.V. All rights reserved.

Background The association between iron overload (IO) and risk of cardiovascular disease is controversial. Epidemiological studies have found a significant negative association of transferrin (Tf) saturation and cardiovascular events suggesting that higher body iron possibly confer a protective effect towards developing cardiovascular events. The biological mechanisms of this phenomenon are unknown. Objective This article investigates the role of IO on platelet reactivity. Materials and Methods This study was a prospective case-control study comparing 45 patients with IO, mostly characterized by the HFE gene mutations C282Y and/or H63D, with 32 healthy controls. We evaluated: (1) platelet aggregation in both platelet-rich plasma and whole blood, (2) platelet membrane expression of the activation marker CD62P, (3) activation of platelet signalling phosphoinositide 3-kinase/Akt and mitogen-activated protein kinase/extracellular signal-regulated kinases (Erk)-1/2 pathways, (4) a pattern of in vivo platelet activation markers, and (5) iron biomarker predictors of platelet reactivity. Results IO patients had significantly lower platelet aggregability, expression of CD62P and phosphorylation amounts of pAkt and pErk-2 in response to agonists. Furthermore, patients with higher Tf saturation levels were characterized by lower circulating levels of sCD40L, PDGF-BB and thromboxane B-2. Platelet aggregation and activation parameters inversely correlated with Tf saturation and the stepwise multivariate regression analysis underlined the role of Tf saturation in predicting platelet reactivity. We also found that in vitro platelet exposure to diferric Tf, but not to iron-depleted TF, dose-dependently inhibited platelet function in all investigated subjects. Conclusion Tf saturation is inversely associated with platelet reactivity and this could explain, at least in part, the association of high Tf and lower risk of cardiovascular diseases in IO.

A diagnosis of rhino-orbital-cerebral mucormycosis was made in a 59-year-old man with a secondary acute myeloid leukemia a few days after hematopoietic stem cell transplantation. Prompt treatment with combined antifungal therapy (liposomal amphotericin B and isavuconazole) followed by a procedure of endoscopic sinus surgery resulted in the resolution of the infection. Therapeutic drug monitoring of isavuconazole was performed during the year of treatment showing an increment of plasma concentrations in correspondence with the improvement of intestinal GvHD, thus suggesting that in this or similar conditions TDM for isavuconazole can be of value. A literature review of cases of rhino-orbital-cerebral and rhino-cerebral mucormycosis in allogeneic hematopoietic stem cell transplant recipients was carried out.

Bone marrow granulomatosis is not a common finding and remains a diagnostic challenge. Here below we report two bizarre but not so rare cases of bone marrow granulomatosis: bone marrow involvement by Mycobacterium avium-intracellulare and reaction to intravesical administration of Bacillus Calmette-Guérin for non-muscle-invasive bladder cancer; two distinct culprits suggesting the wide range of possible causative factors of this histopathological pattern. In the discussion we provide a brief overview of causes and suggestions for a diagnostic approach.

The development of drugs able to target BTK, PI3k-delta and BCL2 has dramatically improved chronic lymphocytic leukaemia (CLL) therapies. However, drug resistance to these therapies has already been reported due to non-recurrent changes in oncogenic pathways and genes expression signatures. In this study, we investigated the cooperative role of the BCL2 inhibitor venetoclax and the BRD4 inhibitor JQ1. In particular, we found that JQ1 shows additional activity with venetoclax, in CLL cell lines and in ex vivo isolated primary CD19+ lymphocytes, arguing in favour of combination strategies. Lastly, JQ1 is also effective in venetoclax-resistant CLL cell lines. Together, our findings indicated that the BET inhibitor JQ1 could be a promising therapy in CLL, both as first-line therapy in combination with venetoclax and as second-line therapy, after the emergence of venetoclax-resistant clones.

Background: Frailty is a common and important geriatric syndrome characterized by age‐associated declines in physiologic reserve and function across multiorgan systems, leading to increased vulnerability for adverse health outcomes. Two major definitions of frailty have emerged over the past years: 1) the frailty phenotype (FP) defined by at least three or five items between weakness, slowness, low physical activity, low energy and weight loss 2) frailty index (FI) based on a comprehensive geriatric assessment (CGA) that evaluates functional status, polypharmacy, comorbidity, emotional and cognitive mental health status, fatigue, socio‐economic condition, nutrition and quality of life. In patients with cancer, the identification of frailty might be relevant to choose the most appropriate therapy for each patient. In hematology the CGA is not routinely performed because it is complex and time‐consuming. Hand Grip strength (HGS) is a general indicator of muscle strength and low HGS has been linked with premature mortality. Aims: The aim of this study was to test HGS as screening tool for frailty in older patients with hematologic malignancies. Methods: Patients ≥ 70 years on treatment for Acute Myeloid Leukemia (AML), Myelodysplastic Syndrome (MDS), Multiple Myeloma (MM), Chronic Linfocytic Leukemia (CLL) and Non Hodgkin Lymphoma (NHL) who were referred to the hematology department of two centers were included in the study. HGS was evaluated with a Jamar dynamometer and CGA was performed with a set of four questionnaires: Activities of Daily Living (ADL), Instrumental Activities of Daily Living (IADL), G8 screening questionnaire, Cumulative Illness Rating Scale (CIRS). Frailty was defined according to the following cut‐off: ADL (>4 ≤4), IADL (>5 ≤5),G8 (>14 ≤14), CIRS ≥1 score 3–4 or > 8 score 2. Spearman's Rank coefficient was used to evaluate the correlation between HGS and questionnaires score. Age and sex‐adjusted logistic regression model was used to evaluate the association between HGS and frailty. The performance of the HGS screening tool was evaluated using Receiver Operating Curve (ROC) analysis and the area under the ROC curve (AUC). Sensitivity and specificity with 95% confidence interval were calculated. Analysis were performed using the software R version 3.5.0. Results: One hundred and eleven patients were included in the study (12 AML 10.8%, 6 MDS 5,4%, 39 MM 35.1%, 11 CLL 9.9% and 43 NHL 38.7%). Median age was 78 ± 4.7 with 42 patients ≥ 80 years (37.8%). The male/female ratio was 66 (59.5%)/45 (40.5%). Overall, HGS significantly correlated with ADL score (r = 0.45 p < 0.001 for men and r = 0.49 p < 0.001 for women), IADL score (r = 0.5 p < 0.001 for men and r = 0.4 p < 0.001 for women), G8 score (r = 0.57 p < 0.05 for men and r = 0.051 p < 0.001 for women) but not with CIRS score. Age and sex‐adjusted logistic regression analysis demonstrated an association between low HGS and frailty in patients with 2 additive scores in ADL, IADL or G8 but not CIRS (OR = 0.021, p = 0.003). In these patients ROC curves of HGS as screening tool for frailty evidenced AUC 87.4% (95%CI: 77.4%>97.4%) in women and 84% (95%CI 72.2%>96.1%) in men. Moreover, the ROC curves revealed an HGS score ≤14 Kg in women (82.1% sensitivity and 70.6% specificity) and ≤25 Kg in men (91.8% sensitivity and 70.6% specificity) as the optimal cut‐off point according to the best Youden index. Summary/Conclusion: HGS is a valid, reliable, rapid and sensible clinical measure of frailty.

Nuclear-cytoplasmic shuttling is a highly regulated and complex process, which involves both proteins and nucleic acids. Changes in cellular compartmentalization of various proteins, including oncogenes and tumor suppressors, affect cellular behavior, promoting or inhibiting proliferation, apoptosis and sensitivity to therapies. In this review, we will recapitulate the role of various shuttling components in Chronic Myeloid Leukemia and we will provide insights on the potential role of shuttling proteins as therapeutic targets.

Aim: Proprotein Convertase Subtilisin/Kexin 9 (PCSK9) plays a major role in the regulation of LDL-receptor function and therapy with PCSK9 inhibitors have shown efficacy in reducing LDL-cholesterol levels whereas their effects on platelets are unknown.

Background: Medical advances in the treatment of Acute and Chronic Leukemia in the last decade were exciting, thanks to the discovery of new drugs and transplantation of hematopoietic stem cells; but so that these weapons can be used in an optimal way, it is necessary that the leukemia is diagnosed as quickly as possible and accurately to limit the spread of leukemia cells in the patient. The ability to detect fusion genes characteristics of these diseases has become essential but laborious, time consuming and generally comprises karyotype analysis, followed by FISH (Fluorescent in situ hybridization) and molecular analysis (RT-PCR). New molecular diagnostic technologies are being developed that can identify disease markers with extreme accuracy in a few hours (2-3 hours). The Leukemia Fusion Genes (Q30) Screening Kit (QuanDx, San Jose, CA 95131 USA; Distr. Resnova S.r.l.) is a real-time multiplex RT-qPCR based assay for detection of leukemia associated fusion gene transcripts in total RNA from bone marrow or whole blood samples, it is a qualitative test for the simultaneous detection of 30 characteristic fusion genes of acute leukemias. For Acute Myeloid Leukemia (AML) transcripts we can expect tested 18 fusion genes with 72 different breakpoints: BCR-ABL1, MLL-AF9, CBFbeta-MYH11, PML-RARalpha, AML-MDS1, NPM-MLF1, AML-ETO, NPM-RARalpha, PLZF-RARalpha, DEK-CAN, MLL-ELL, AML1-EAP, MLL-AF10, ST-CAN, TEL-ABL1, TLS-ERG, FIP1L1-PDGFR alpha, TEL-PDGFRbeta. For Acute Lymphoid Leukemia (ALL) transcripts we will detected 15 fusion genes with 71 different breakpoints: MLL-AF4, TEL-AML1, MLL-ENL, E2APBX1, BCR/ABL1, SIL-TAL1, E2A-HLF, MLL-AF6, CALM-AF10, HOX11, HOX11L2, MLL-AF10, HOX11, HOX11L2, MLL-AF10, SET-CAN, TEL-ABL1, TLS-ERG.

Summary Glucagon-like peptide 1 (GLP-1) is object of intensive investigation for not only its metabolic effects but also the protective vascular actions. Since platelets exert a primary role in the pathogenesis of atherosclerosis, inflammation and vascular complications, we investigated whether GLP-1 directly influences platelet reactivity. For this purpose, in platelets from 72 healthy volunteers we evaluated GLP-1 receptor (GLP-1R) expression and the effects of a 15-minute incubation with the native form GLP-1(7–36), the N-terminally truncated form GLP-1(9–36) and the GLP-1 analogue Liraglutide (100 nmol/l) on: i) aggregation induced by collagen or arachidonic acid (AA); ii) platelet function under shear stress; iii) cGMP and cAMP synthesis and cGMP-dependent protein kinase (PKG)-induced Vasodilator-Stimulated-Phosphoprotein (VASP) phosphorylation; iv) activation of the signalling molecules Phosphatidylinositol 3-Kinase (PI3-K)/Akt and Mitogen Activated Protein Kinase (MAPK)/ERK-1/2; and v) oxidative stress. Experiments were repeated in the presence of the nitric oxide donor Na–nitroprusside. We found that platelets constitutively express GLP-1R and that, independently of GLP-1R, GLP-1(7–36), GLP-1(9–36) and Liraglutide exert platelet inhibitory effects as shown by: a) increased NO-antiaggregating effects, b) increased the activation of the cGMP/PKG/VASP pathway, c) reduced the activation of PI3-K/Akt and MAPK/ERK-2 pathways, d) reduced the AA-induced oxidative stress. When the experiments were repeated in the presence of the antagonist of GLP-1R Exendin(9–39), the platelet inhibitory effects were maintained, thus indicating a mechanism independent of GLP-1R. In conclusion, GLP-1(7–36), its degradation product GLP-1(9–36) and Liraglutide exert similar inhibitory effects on platelet activation, suggesting a potential protective effect on the cardiovascular system.

Chronic Lymphocytic Leukemia (CLL) is a lymphoproliferative disorder with either indolent or aggressive clinical course. Current treatment regiments have significantly improved the overall outcomes even if higher risk subgroups -those harboring TP53 mutations or deletions of the short arm of chromosome 17 (del17p) -remain highly challenging. In the present work, we identified USP7, a known de-ubiquitinase with multiple roles in cellular homeostasis, as a potential therapeutic target in CLL. We demonstrated that in primary CLL samples and in CLL cell lines USP7 is: i) overexpressed through a mechanism involving miR-338-3p and miR-181b deregulation; ii) functionally activated by Casein Kinase 2 (CK2), an upstream interactor known to be deregulated in CLL; iii) effectively targeted by the USP7 inhibitor P5091. Treatment of primary CLL samples and cell lines with P5091 induces cell growth arrest and apoptosis, through the restoration of PTEN nuclear pool, both in TP53-wild type and -null environment. Importantly, PTEN acts as the main tumor suppressive mediator along the USP7-PTEN axis in a p53 dispensable manner. In conclusion, we propose USP7 as a new druggable target in CLL.