Estrogen-dependent dynamic profile of eNOS-DNA associations in prostate cancer

In previous work we have documented the nuclear translocation of endothelial NOS (eNOS) and its participation in combinatorial complexes with Estrogen Receptor Beta (ERβ) and Hypoxia Inducible Factors (HIFs) that determine localized chromatin remodeling in response to estrogen (E2) and hypoxia stimuli, resulting in transcriptional regulation of genes associated with adverse prognosis in prostate cancer (PCa). To explore the role of nuclear eNOS in the acquisition of aggressive phenotype in PCa, we performed ChIP-Sequencing on chromatin-associated eNOS from cells from a primary tumor with poor outcome and from metastatic LNCaP cells. We found that: 1. the eNOS-bound regions (peaks) are widely distributed across the genome encompassing multiple transcription factors binding sites, including Estrogen Response Elements. 2. E2 increased the number of peaks, indicating hormone-dependent eNOS re-localization. 3. Peak distribution was similar with/without E2 with ≈ 55% of them in extragenic DNA regions and an intriguing involvement of the 5′ domain of several miRs deregulated in PCa. Numerous potentially novel eNOS-targeted genes have been identified suggesting that eNOS participates in the regulation of large gene sets. The parallel finding of downregulation of a cluster of miRs, including miR-34a, in PCa cells associated with poor outcome led us to unveil a molecular link between eNOS and SIRT1, an epigenetic regulator of aging and tumorigenicity, negatively regulated by miR-34a and in turn activating eNOS. E2 potentiates miR-34a downregulation thus enhancing SIRT1 expression, depicting a novel eNOS/SIRT1 interplay fine-tuned by E2-activated ER signaling, and suggesting that eNOS may play an important role in aggressive PCa

Realizzazione di un nuovo sito web per l’INAF - Istituto di Radioastronomia

Questo report illustra l'attivita` del gruppo di lavoro incaricato della progettazione e sviluppo del nuovo sito web dell'INAF - Istituto di Radioastronomia. L'utilizzo di tecnologie di sviluppo web basate su CMS ha consentito di superare le difficolta` nell'aggiornamento e gestione del precedente sito basato su pagine html statiche. Con il nuovo approccio si è ottenuto da un lato un notevole miglioramento dell'aspetto grafico e della coerenza tra le pagine, dall’altro una notevole semplificazione nella produzione e nell’aggiornamento delle pagine stesse. Cio` consentira` ai diversi gruppi di ricerca, servizi e uffici di farsi carico delle pagine di competenza e di poter facilmente apportare modifiche ai contenuti via via che queste si renderanno necessarie, garantendo allo stesso tempo una maggiore sicurezza informatica. Nel report vengono descritte le varie fasi dell'attivita` del gruppo di lavoro: dalla iniziale identificazione delle linee guida e progettazione di struttura e contenuti, sino alla realizzazione vera e propria delle pagine web e ai successivi step di verifica conclusi con la pubblicazione online del sito

Myc and Omomyc functionally associate with the Protein Arginine Methyltransferase 5 (PRMT5) in glioblastoma cells

The c-Myc protein is dysregulated in many human cancers and its function has not been fully elucitated yet. The c-Myc inhibitor Omomyc displays potent anticancer properties in animal models. It perturbs the c-Myc protein network, impairs c-Myc binding to the E-boxes, retaining transrepressive properties and inducing histone deacetylation. Here we have employed Omomyc to further analyse c-Myc activity at the epigenetic level. We show that both Myc and Omomyc stimulate histone H4 symmetric dimethylation of arginine (R) 3 (H4R3me2s), in human glioblastoma and HEK293T cells. Consistently, both associated with protein Arginine Methyltransferase 5 (PRMT5)-the catalyst of the reaction-and its co-factor Methylosome Protein 50 (MEP50). Confocal experiments showed that Omomyc co-localized with c-Myc, PRMT5 and H4R3me2s-enriched chromatin domains. Finally, interfering with PRMT5 activity impaired target gene activation by Myc whereas it restrained Omomyc-dependent repression. The identification of a histone-modifying complex associated with Omomyc represents the first demonstration of an active role of this miniprotein in modifying chromatin structure and adds new information regarding its action on c-Myc targets. More importantly, the observation that c-Myc may recruit PRMT5-MEP50, inducing H4R3 symmetric di-methylation, suggests previously unpredictable roles for c-Myc in gene expression regulation and new potential targets for therapy

Telomerase Mediates Vascular Endothelial Growth Factor-dependent Responsiveness in a Rat Model of Hind Limb Ischemia *

Telomere dysfunction contributes to reduced cell viability, altered differentiation, and impaired regenerative/proliferative responses. Recent advances indicate that telomerase activity confers a pro-angiogenic phenotype to endothelial cells and their precursors. We have investigated whether telomerase contributes to tissue regeneration following hind limb ischemia and vascular endothelial growth factor 165 (VEGF(165)) treatment. VEGF delivery induced angiogenesis and increased expression of the telomerase reverse transcriptase (TERT) and telomerase activity in skeletal muscles and satellite and endothelial cells. Adenovirus-mediated transfer of wild type TERT but not of a dominant negative mutant, TERTdn, significantly induced capillary but not arteriole formation. However, when co-delivered with VEGF, TERTdn abrogated VEGF-dependent angiogenesis, arteriogenesis, and blood flow increase. This effect was paralleled by in vitro evidence that telomerase inhibition by 3'-azido-3'-deoxythymidine in VEGF-treated endothelial cells strongly reduced capillary density and promoted apoptosis in the absence of serum. Similar results were obtained with adenovirus-mediated expression of TERTdn and AKTdn, both reducing endogenous TERT activity and angiogenesis on Matrigel. Mechanistically, neo-angiogenesis in our system involved: (i) VEGF-dependent activation of telomerase through the nitric oxide pathway and (ii) telomerase-dependent activation of endothelial cell differentiation and protection from apoptosis. Furthermore, detection of TERT in activated satellite cells identified them as VEGF targets during muscle regeneration. Because TERT behaves as an angiogenic factor and a downstream effector of VEGF signaling, telomerase activity appears required for VEGF-dependent remodeling of ischemic tissue at the capillaries and arterioles level

Radio data archiving system

Radio Astronomical Data models are becoming very complex since the huge possible range of instrumental configurations available with the modern Radio Telescopes. What in the past was the last frontiers of data formats in terms of efficiency and flexibility is now evolving with new strategies and methodologies enabling the persistence of a very complex, hierarchical and multi-purpose information. Such an evolution of data models and data formats require new data archiving techniques in order to guarantee data preservation following the directives of Open Archival Information System and the International Virtual Observatory Alliance for data sharing and publication. Currently, various formats (FITS, MBFITS, VLBI's XML description files and ancillary files) of data acquired with the Medicina and Noto Radio Telescopes can be stored and handled by a common Radio Archive, that is planned to be released to the (inter)national community by the end of 2016. This state-of-the-art archiving system for radio astronomical data aims at delegating as much as possible to the software setting how and where the descriptors (metadata) are saved, while the users perform user-friendly queries translated by the web interface into complex interrogations on the database to retrieve data. In such a way, the Archive is ready to be Virtual Observatory compliant and as much as possible user-friendly

A public data archive for the Italian radio telescopes

The amount of data delivered by modern instrumentation and observing techniques is bringing radio astronomy in the era of Big Data, and the nowadays widely adopted Open Data policies allow free and open access to data from many radio astronomy facilities. A fundamental ingredient to enable Open Science in the radio astronomical community and to engage also public participation (the so called Citizen Science) is thus the availability of public archives in which data can be accessed and searched with modern software tools. A web-based, VO-compliant public archive has been built to host data from the Italian radio telescopes managed by the National Institute for Astrophysics (INAF). The archive main features consist in the capability to handle the various types of data coming from the different observing instrumentation at the telescopes; the adoption of a policy to guarantee the data proprietary period; the accessibility of data through a web interface and the adoption of VO standards to allow for successful scientific exploitation of the archive itself in the data mining era. We present the progress status of the public Data Archive for the Italian radio telescopes being developed to provide the international community with a state-of-the-art archive for radio astronomical data

Sildenafil normalizes MALAT1 level in diabetic cardiomyopathy

Introduction A large body of evidence recently highlighted the involvement of long non-coding RNAs (lncRNAs) in cardiovascular disease [1] and some dysregulated lncRNAs have been associated with diabetic cardiomyopathy [2\u20135]. Among them, a higher expression of the lncRNA metastasis associated lung adenocarcinoma transcript 1 (MALAT1) has been observed in diabetic cardiomyopathy [6, 7]. However, a clear understanding of the molecular mechanisms leading to pathological regulation of lncRNAs in diabetic cardiomyopathy is still missing. Our prior work by Barbati et al. [8], established that, in the presence of high glucose, nitric oxide (NO) signaling derangement might alter the epigenetic landscape of cardiac cells, both in vitro and in vivo, via transcription factor CREM activation. Aim The present study is aimed at investigating the role of high glucose (HG) and NO pathway in the regulation of MALAT1 in the heart of mice after 6 months of prolonged hyperglycemia and in two cellular models of cardiomyocytes exposed to HG

MALAT1 as a Regulator of the Androgen-Dependent Choline Kinase A Gene in the Metabolic Rewiring of Prostate Cancer

Simple Summary Despite the rapid advance in cancer therapies, treatment-resistant relapse remains a significant challenge in cancer treatment. Acquired resistance arises during or after treatment administration, and is usually the main contributor to relapse. For example, prostate cancer, the most frequent type of cancer in the elderly male population, frequently develops into aggressive forms resistant to chemical and hormonal therapies. In this condition, the so-called "cholinic phenotype" that is characterized by the overexpression of choline kinase alpha (CHKA) and increased phosphocholine levels leads to aberrant lipid metabolism. Our work demonstrates that CHKA, which is necessary for membrane phospholipid synthesis, is a target of the long non-coding RNA MALAT1. This study helps to further decipher how MALAT1 affects the regulation of crucial phospholipid/sphingolipid metabolic enzymes, as well as how the androgen receptor pathway is involved in MALAT1-dependent transcriptional regulation. Background. Choline kinase alpha (CHKA), an essential gene in phospholipid metabolism, is among the modulated MALAT1-targeted transcripts in advanced and metastatic prostate cancer (PCa). Methods. We analyzed CHKA mRNA by qPCR upon MALAT1 targeting in PCa cells, which is characterized by high dose-responsiveness to the androgen receptor (AR) and its variants. Metabolome analysis of MALAT1-depleted cells was performed by quantitative High-resolution 1 H-Nuclear Magnetic Resonance (NMR) spectroscopy. In addition, CHKA genomic regions were evaluated by chromatin immunoprecipitation (ChIP) in order to assess MALAT1-dependent histone-tail modifications and AR recruitment. Results. In MALAT1-depleted cells, the decrease of CHKA gene expression was associated with reduced total choline-containing metabolites compared to controls, particularly phosphocholine (PCho). Upon MALAT1 targeting a significant increase in repressive histone modifications was observed at the CHKA intron-2, encompassing relevant AR binding sites. Combining of MALAT1 targeting with androgen treatment prevented MALAT1-dependent CHKA silencing in androgen-responsive (LNCaP) cells, while it did not in hormone-refractory cells (22RV1 cells). Moreover, AR nuclear translocation and its activation were detected by confocal microscopy analysis and ChIP upon MALAT1 targeting or androgen treatment. Conclusions. These findings support the role of MALAT1 as a CHKA activator through putative association with the liganded or unliganded AR, unveiling its targeting as a therapeutic option from a metabolic rewiring perspective

Chandra and Magellan/FIRE follow-up observations of PSO167-13: an X-ray weak QSO at z=6.515z=6.515

The discovery of hundreds of QSOs in the first Gyr of the Universe powered by already grown SMBHs challenges our knowledge of SMBH formation. In particular, investigations of $z>6$ QSOs presenting notable properties can provide unique information on the physics of fast SMBH growth in the early universe. We present the results of follow-up observations of the $z=6.515$ radio-quiet QSO PSO167-13, which is interacting with a close companion galaxy. The PSO167-13 system has been recently proposed to host the first heavily obscured X-ray source at high redshift. We observed PSO167-13 with Chandra/ACIS-S (177 ks), and obtained new spectroscopic observations (7.2 h) with Magellan/FIRE. No significant X-ray emission is detected from the PSO167-13 system, suggesting that the obscured X-ray source previously tentatively detected was either due to a strong background fluctuation or is highly variable. The upper limit (90% confidence level) on the X-ray emission of PSO167-13 ($L_{2-10\,\mathrm{keV}}<8.3\times10^{43}\,\mathrm{erg s^{-1}}$) is the lowest available for a $z>6$ QSO. The ratio between the X-ray and UV luminosity of $\alpha_{ox}<-1.95$ makes PSO167-13 a strong outlier from the $\alpha_{ox}-L_{UV}$ and $L_X-L_{\mathrm{bol}}$ relations. In particular, its X-ray emission is $>6$ times weaker than the expectation based on its UV luminosity. The new Magellan/FIRE spectrum of PSO167-13 is strongly affected by the unfavorable sky conditions, but the tentatively detected C IV and Mg II emission lines appear strongly blueshifted. The most plausible explanations for the X-ray weakness of PSO167-13 are intrinsic weakness or small-scale absorption by Compton-thick material. The possible strong blueshift of its emission lines hints at the presence of nuclear winds, which could be related to its X-ray weakness.Comment: Accepted for publication on A&