Investigation of the oncogenic role of MAF in multiple myeloma

Multiple myeloma (MM) is an incurable malignancy of bone marrow plasma cells (PC), affecting over 5,600 new patients per year in the UK. Translocation of the transcription factor MAF to the IgH enhancer, t(14;16), is a myeloma-initiating event in 3-5% of MM cases. As well as in t(14;16), MAF is over-expressed in another 30% of MM, mostly in association with the MMSET t(4;14) translocation. The mechanisms by which aberrant expression of MAF promotes myelomagenesis, including the oncogenic transcriptome it regulates as well as the regulatory regions it impacts are unknown. In this work, with reference to the normal PC, and using both primary myeloma cells and myeloma cell lines, I have applied a multi-omics approach to define the MAF regulome and determine the impact of ectopic MAF expression on MM cells. First, analysis of the MAF transcriptome in primary myeloma, as compared to normal PC, revealed upregulated genes that are shared with other MM subgroups or are MAF-unique. Having established MAF as a t(14;16) myeloma cell dependency using knock down assays, I obtained and integrated the transcriptome following MAF depletion with the MAF cistrome using RNA-seq and ChIP-seq respectively. This identified 2,112 genes directly bound and regulated by MAF, the majority of which were predicted to be activated by the transcription factor. Intersecting these direct MAF targets with the genes differentially upregulated in MAF primary MM cells, revealed a core transcriptional MAF programme involved in cell cycle regulation and cell to cell interactions such as cell adhesion, migration, cytokine and chemokine signaling pathways. Identification of the MAF cistrome revealed that MAF preferentially binds enhancers, prompting further exploration of the MAF-associated distal regulatory genome. Compared to normal PC, t(14;16) primary MM cells displayed enhanced chromatin accessibility as assessed by ATAC-seq, with 33% of those regions being over-accessible uniquely in the MAF MM subgroup. Indeed, the chromatin accessibility profile of MAF MM was able to distinctly cluster the samples separately from other genetic MM groups. Remarkably, a quarter of the over-accessible MAF regions were also bound by the MAF (in total 1,661 regions), consistent with a role of MAF in the regulation of chromatin accessibility. Juxtaposing these regions against chromatin mark maps of the MAF translocated MM.1S cell line and normal PC revealed 44% (726/1,661) of these regions, acquire activatory marks in MAF-translocated cells having been inactive in normal PC and the majority also inactive throughout B cell development. This finding suggested the possible implication of MAF in the activation of these regions. Indeed, following overexpression of MAF in a MAF-negative myeloma cell line, over 50% of these regions acquired enhanced accessibility, demonstrating MAF as an activator of specific regulatory regions in t(14;16) MM. This enabled identification of MAF-activated enhancers, such as in the case of a CCR1 enhancer peak, validated to be a CCR1 regulatory region by CRISPRi. Finally, using ChIP-seq data and chromatin proteomics I find IRF4 to be co-bound in 70-80% of MAF-bound genomic regions, suggesting possible cooperation as well as cross-regulation between the two transcription factors. In addition, several other proteins are identified as candidate partners of the MAF interactome. In conclusion, this work has provided a comprehensive description of the role of oncogenic MAF in MM and given insights into events related to ectopic MAF expression in PC occurring during the process of myelomagenesis. It has revealed the full spectrum of directly MAF-regulated genes and the consequent cellular pathways affected in MM. In addition, it has established MAF’s role in modifying the PC regulatory genome by activating specific regulatory regions, and has enabled the identification of enhancers regulating key MAF-dependent genes, critical for myeloma cell biology.Open Acces

The discrete ability of the interaction of psychological factors and dierary habits in the assessing the likelihood of having a first acute acute coronary syndrome or stroke: a case control study

Atherosclerotic cardiovascular disease remains the leading cause of premature death in Europe, although cardiovascular mortality has been markedly decreased across European countries during the last decades. Except from the already known and thoroughly documented traditional cardiovascular disease risk factors (i.e. central adiposity, smoking, low Mediterranean diet adherence etc), research interest focuses on novel risk factors and their role in coronary heart disease manifestation is examined. For instance, several biopsychosocial factors (i.e. depression, anxiety, anger, hostility, work stress and stress in general) have been studied for their role in cardiovascular disease development and onset. Actually, perceived stress presents a global, comprehensive and integrated framework, but its role in coronary heart disease incidence is not clear yet. The aim of the current work was the evaluation of the independent role of perceived stress on the likelihood of having an ACS. Perceived stress was measured by the PSS-14. Moreover, any potential mediating or moderating role of the Mediterranean diet adherence, as well as certain stress related eating behaviors (i.e. eating alone, stress eating, and eating large meals) on the relationship between perceived stress and CVD incidence was also evaluated. A Greek version PSS-14 was validated in a sample of adults from the general population in order to allow the conduct of the work. In the current study 250 patients with a first ACS event and without any suspicion of previous CVD (60±11 years old, 78% men), as well as 250 healthy individuals, were enrolled (60±8.6 years old, 77.6% men). Perceived Stress was assessed using the Perceived Stress Scale, a 14-item self-reported questionnaire (range 0-56) and adherence to the Mediterranean diet was evaluated using an 11-item large-scale, composite index, the MedDietScore (range 0-55). Eating behaviors, such as stress eating, eating large and heavy meals and eating alone were examined as well. PSS-14 was positively associated (OR: 1.147, 95%CI 1.11-1.18) with the likelihood of having an ACS even after adjusting for several related variables. Moreover, each unit of increase of the MedDietScore led to a 54% lower likelihood of having an ACS (95%CI 0.27-0.79). Sensitivity analysis by level of adherence to the Mediterranean diet revealed a similar association between perceived stress and the likelihood of having an ACS in the low adherence group (OR=1.15, 95%CI 1.09, 1.21) and moderate-to-high adherence group respectively (OR=1.13, %CI 1.07, 1.80). Stress eating and eating heavy meals were positively associated with the likelihood of having an ACS (OR=1.91, 95%CI 1.07, 3.41 and OR=5.30, 95%CI 2.97, 9.45 respectively).The current study confirmed the relationship between perceived stress and presence of ACS with the use of PSS-14. This significant impact of perceived stress on the likelihood of having an ACS was not mediated nor moderated by the level of Mediterranean diet adherence or other eating behaviors, albeit the documented effect of this particular psychological disposition on the likelihood of having an ACS. In a public health perspective, it is suggested that levels of perceived stress should not remain undiagnosed in populations that may be characterized by a potential cardiovascular risk. Additionally, higher adherence to Mediterranean diet principles, irrespective of stress levels, shall be emphasized. Further, the independent role – irrespective of stress related behaviors - of Mediterranean diet on cardiovascular risk may present a novel research hypothesis, as well.Η αθηροσκληρωτική καρδιαγγειακή νόσος παραμένει η σοβαρότερη αιτία πρόωρου θανάτου στην Ευρώπη, ακόμα και αν η καρδιαγγειακή θνησιμότητα έχει μειωθεί αξιοσημείωτα στις δεκαετίες στις περισσότερες Ευρωπαϊκές χώρες. Πέρα από τους κλασσικούς και εξαιρετικά μελετημένους καρδιαγγειακούς παράγοντες κινδύνου (π.χ. κεντρική παχυσαρκία, κάπνισμα, διατροφή μακριά από το μεσογειακό πρότυπο κ.ά.), νεώτεροι παράγοντες διερευνώνται επίσης ως προς το ρόλο τους στην ανάπτυξη και την εκδήλωση στεφανιαίας νόσου. Για παράδειγμα, διάφοροι ψυχοκοινωνικοί παράγοντες (π.χ. κατάθλιψη, άγχος, θυμός, επιθετικότητα, η εργασιακή ψυχοσωματική φόρτιση (εργασιακό στρες) και, γενικότερα, η ψυχοσωματική φόρτιση (στρες) έχουν διερευνηθεί αρκετά για το ρόλο τους στην ανάπτυξη και την εκδήλωση της στεφανιαίας νόσου. Η εξατομικευμένη συνολική ψυχοσωματική αντίδραση του κάθε οργανισμού σε ερεθίσματα του εξωτερικού ή/και εσωτερικού περιβάλλοντος (στη βιβλιογραφία ο όρος αναφέρεται ως perceived stress, υποδηλώνοντας την ψυχοσωματική φόρτιση που γίνεται αντιληπτή από το άτομο) αποτελεί μία σφαιρική, ολοκληρωμένη και περιεκτική έννοια αλλά ο ρόλος του στην επίπτωση της στεφανιαίας νόσου δεν έχει καταστεί ξεκάθαρος ακόμη. Σκοπό της παρούσας μελέτης ασθενών-μαρτύρων αποτέλεσε η εκτίμηση της ανεξάρτητης φύσης της εξατομικευμένης ψυχοσωματικής φόρτισης (αντιληπτού στρες) ως καρδιαγγειακού παράγοντα κινδύνου, μετρημένου με το εργαλείο – ερωτηματολόγιο PSS-14, στην πιθανότητα εμφάνισης οξέος εμφράγματος του μυοκαρδίου. Επιπλέον, διερευνήθηκε ο πιθανός διαμεσολαβητικός ή/και τροποποιητικός ρόλος της υιοθέτησης της Μεσογειακής διατροφής, αλλά και διατροφικών συμπεριφορών που σχετίζονται με την ψυχοσωματική φόρτιση (στρες) (π.χ. κατανάλωση τροφής χωρίς παρέα, υπό πίεση ή κατανάλωση μεγάλων γευμάτων που προκαλούν δυσφορία), στη σχέση μεταξύ της εξατομικευμένης ψυχοσωματικής φόρτισης (αντιληπτού στρες) και της πιθανότητας εμφάνισης οξέος εμφράγματος του μυοκαρδίου. Μάλιστα, για την εξυπηρέτηση του σκοπού της παρούσας εργασίας, εκτιμήθηκε η εγκυρότητα της ελληνικής έκδοσης του PSS-14 σε ένα δείγμα ενηλίκων από τον ελληνικό γενικό πληθυσμό. Στη μελέτη συμμετείχαν 250 ασθενείς με πρωτοεμφανιζόμενη στεφανιαία νόσο (60±11 ετών, 78% άνδρες) και 250 υγιείς (60±8.6 ετών, 77.6% άνδρες). Τα επίπεδα εξατομικευμένης ψυχοσωματικής φόρτισης (αντιληπτού στρες) εκτιμήθηκαν με το ερωτηματολόγιο PSS-14 (εύρος 0-56) και το επίπεδο υιοθέτησης της Μεσογειακής διατροφής εκτιμήθηκε με το Μεσογειακό διατροφικό σκορ (εύρος 0-55). Διερευνήθηκαν επίσης διατροφικές συμπεριφορές όπως η κατανάλωση τροφής υπό πίεση (στρες), η κατανάλωση βαριών γευμάτων και η κατανάλωση τροφής χωρίς συνδαιτυμόνες. Το PSS-14 συσχετίστηκε θετικά με την πιθανότητα εμφάνισης οξέος στεφανιαίου συνδρόμου και ειδικότερα, για κάθε μία μονάδα αύξησης του σκορ του PSS-14 (95%ΔΕ 1.11-1.18), η πιθανότητα ύπαρξης οξέος εμφράγματος του μυοκαρδίου αυξήθηκε κατά 15%, ακόμα και μετά την προσαρμογή για σχετικές κλινικές παραμέτρους. Επίσης, για κάθε μονάδα αύξησης στο βαθμό υιοθέτησης της Μεσογειακής Διατροφής, η πιθανότητα παρουσίας οξέος εμφράγματος μειωνόταν κατά 54% (95%ΔΕ 0.27-0.79). Η ανάλυση ευαισθησίας ανά επίπεδο υιοθέτησης της μεσογειακής δίαιτας αποκάλυψε παρόμοια συσχέτιση μεταξύ εξατομικευμένης ψυχοσωματικής φόρτισης (αντιληπτού στρες) και παρουσίας ΟΕΜ μεταξύ της ομάδας χαμηλής υιοθέτησης και της ομάδας μέτριας-υψηλής (π.χ. ΣΛ=1.15, 95%ΔΕ 1.09, 1.21, και ΣΛ=1.13, 95%ΔΕ 1.07, 1.80, αντίστοιχα). Η κατανάλωση τροφής υπό στρες και η κατανάλωση βαριών γευμάτων συσχετίστηκαν επίσης με την παρουσία ΟΣΣ (ΣΛ=1.91, 95%ΔΕ 1.07, 3.41 και ΣΛ=5.30, 95%ΔΕ 2.97, 9.45 αντίστοιχα).Η παρούσα εργασία επιβεβαίωσε τη σχέση της εξατομικευμένης ψυχοσωματικής φόρτισης (αντιληπτού στρες) και της παρουσίας ΟΕΜ με τη χρήση του εργαλείου PSS-14. Η ισχυρά σημαντική επίδραση της εξατομικευμένης ψυχοσωματικής φόρτισης (αντιληπτού στρες) στην πιθανότητα εμφάνισης ΟΕΜ δεν διαμεσολαβήθηκε ή τροποποιήθηκε από το επίπεδο υιοθέτησης της μεσογειακής δίαιτας ή από άλλες διατροφικές συμπεριφορές, δεδομένης της ισχυρής επίδρασης αυτής της ψυχολογικής διαταραχής στην πιθανότητα εμφάνισης ΟΕΜ. Τα επίπεδα εξατομικευμένης ψυχοσωματικής φόρτισης (αντιληπτού στρες) δε θα πρέπει να παραμένουν αδιάγνωστα σε πληθυσμούς με πιθανό καρδιαγγειακό κίνδυνο. Επιπλέον, μεγαλύτερη υιοθέτηση στις αρχές της μεσογειακής διατροφής, ανεξάρτητα από τις στρεσογόνες εμπειρίες του ατόμου, δικαιούται κατάλληλης έμφασης και μπορεί να αποτελεί άλλη μία ερευνητική υπόθεση σχετικά με τον - ανεξάρτητο από στρεσογόνες συμπεριφορές - ρόλο της μεσογειακής δίαιτας στον καρδιαγγειακό κίνδυνο

Effect of sodium valproate monotherapy on serum uric acid concentrations in ambulatory epileptic children: A prospective long-term study

Purpose: Hyperuricemia has been shown to be related to cardiovascular morbidity and mortality. There is controversial data about the effect of sodium valproate (VPA) monotherapy on serum uric acid concentrations. The purpose of this study was to investigate by a long-term, prospective method, whether treatment with VPA monotherapy may alter serum uric acid concentrations and liver function tests in ambulatory epileptic children. Material and methods: Serum uric acid concentrations were determined in 28 ambulatory epileptic children before and at 6, 12 and 24 months of VPA monotherapy. Serum concentrations of biochemical markers of liver and renal function, such as alanine aminotransferase (ALT), aspartate aminotransferase (AST), lactate dehydrogenase (LDH), gamma-glutamyltransferase (gamma-GT) and creatinine (Cr) were also measured before and at 6, 12 and 24 months of VPA monotherapy. Serum VPA concentrations remained within the therapeutic range (50-100 mg/L) during the period of study. Results: No statistically significant changes in serum Uric acid concentrations were found at 6, 12 or 24 months of treatment. Serum ALT concentrations were significantly increased at 6 and 12 months of treatment, AST concentrations at 6 and 12 months of treatment and LDH concentrations at 12 months of treatment. Conclusions: VPA monotherapy does not have a significant effect on serum uric acid concentrations in ambulatory epileptic children. Further studies are needed to definitively address whether it would be useful for physicians to routinely check for elevated serum uric acid levels in children treated with VPA. (C) 2006 European Paediatric Neurology Society. Published by Elsevier Ltd. All rights reserved

Stress management and dietary counseling in hypertensive patients: a pilot study of additional effect

Background: In Western societies, cardiovascular (CV) disease is the primary cause of mortality, and high blood pressure (BP) is the main reversible factor leading to CV disease. Dietary habits and psychosocial stress contribute to the establishment of hypertension, while its role in the control of high BP is currently examined. In this study, we examined the effect and feasibility of a combined intervention of dietary education and stress management on the control of hypertension. Methodology: A randomized, controlled pilot study was designed to evaluate the effect of combined education on stress management techniques and dietary habits (Mediterranean diet principle) on office BP after eight weeks. Results: Of the 45 randomized subjects, 36 were included in the final analysis (control group = 20 (age: 67 +/- 12 years, 31.8%, males) and intervention group516 (age: 62 +/- 12 years, 47%, males)). CV disease risk factors (except smoking), BP, dietary habits, perceived stress and physical activity (all assessed with validated questionnaires) were similar between the two groups at baseline. After eight weeks, office BP (systolic and diastolic) and perceived stress were significantly reduced, whereas the adherence in Mediterranean diet principle was significantly increased, but only in the intervention group. Conclusions: A combined intervention of stress management techniques and Mediterranean diet education seems to be beneficial for BP reduction. Such interventions could possibly serve as a complementary treatment along with drug therapy or in the early treatment of high normal BP. A call to action for designing epidemiological studies and evaluating the efficacy of such non-pharmacological treatment strategies is therefore warranted

Increased body mass and depressive symptomatology are associated with hypercholesterolemia, among elderly individuals; results from the MEDIS study

Abstract Background Hypercholesterolemia is one of the most important factors causing cardiovascular disease (CVD). The aim of the present work was to evaluate the relationships between socio-demographic, clinical, lifestyle and depression status and the presence of hypercholesterolemia, among elderly individuals without known CVD. Methods During 2005–2007, 1190 elderly (aged 65 to 100 years) men and women (from Cyprus, Mitilini, Samothraki, Cephalonia, Crete, Lemnos, Corfu and Zakynthos) were enrolled. Socio-demographic, clinical and lifestyle factors were assessed through standard procedures. Symptoms of depression were evaluated using the short-form of the Geriatric Depression Scale (GDS, range 0–15). Dietary habits were assessed through a semi-quantitative food frequency questionnaire. Hypercholesterolemia was defined as total serum cholesterol > 200 mg/dL or use of lipids lowering medication. Results 44.6% of males and 61.9% of females had hypercholesterolemia (p Conclusion A considerable proportion of our elderly sample had hypercholesterolemia, while 1/3 of them were untreated. Furthermore, presence of hypercholesterolemia was correlated with depressive symptomatology and increased BMI.</p