Valentina Russo

Cancer-associated fibroblasts release exosomal microRNAs that dictate an aggressive phenotype in breast cancer.

Cancer-associated fibroblasts (CAFs) are the major components of the tumor microenvironment. They may drive tumor progression, although the mechanisms involved are still poorly understood. Exosomes have emerged as important mediators of intercellular communication in cancer. They mediate horizontal transfer of microRNAs (miRs), mRNAs and proteins, thus affecting breast cancer progression. Differential expression profile analysis identified three miRs (miRs -21, -378e, and -143) increased in exosomes from CAFs as compared from normal fibroblasts. Immunofluorescence indicated that exosomes may be transferred from CAFs to breast cancer cells, releasing their cargo miRs. Breast cancer cells (BT549, MDA-MB-231, and T47D lines) exposed to CAF exosomes or transfected with those miRs exhibited a significant increased capacity to form mammospheres, increased stem cell and epithelial-mesenchymal transition (EMT) markers, and anchorage-independent cell growth. These effects were reverted by transfection with anti-miRs. Similarly to CAF exosomes, normal fibroblast exosomes transfected with miRs -21, -378e, and -143 promoted the stemness and EMT phenotype of breast cancer cells. Thus, we provided evidence for the first time of the role of CAF exosomes and their miRs in the induction of the stemness and EMT phenotype in different breast cancer cell lines. Indeed, CAFs strongly promote the development of an aggressive breast cancer cell phenotype.

Aptamer-miRNA-212 Conjugate Sensitizes NSCLC Cells to TRAIL.

TNF-related apoptosis-inducing ligand (TRAIL) is a promising antitumor agent for its remarkable ability to selectively induce apoptosis in cancer cells, without affecting the viability of healthy bystander cells. The TRAIL tumor suppressor pathway is deregulated in many human malignancies including lung cancer. In human non-small cell lung cancer (NSCLC) cells, sensitization to TRAIL therapy can be restored by increasing the expression levels of the tumor suppressor microRNA-212 (miR-212) leading to inhibition of the anti-apoptotic protein PED/PEA-15 implicated in treatment resistance. In this study, we exploited a previously described RNA aptamer inhibitor of the tyrosine kinase receptor Axl (GL21.T) expressed on lung cancer cells, as a means to deliver miR-212 into human NSCLC cells expressing Axl. We demonstrate efficient delivery of miR-212 following conjugation of the miR to GL21.T (GL21.T-miR212 chimera). We show that the chimera downregulates PED and restores TRAIL-mediate cytotoxicity in cancer cells. Importantly, treatment of Axl+ lung cancer cells with the chimera resulted in (i) an increase in caspase activation and (ii) a reduction of cell viability in combination with TRAIL therapy. In conclusion, we demonstrate that the GL21.T-miR212 chimera can be employed as an adjuvant to TRAIL therapy for the treatment of lung cancer.

MiR-221 promotes stemness of breast cancer cells by targeting DNMT3b

Cancer stem cells (CSCs) are a small part of the heterogeneous tumor cell population possessing self-renewal and multilineage differentiation potential as well as a great ability to sustain tumorigenesis. The molecular pathways underlying CSC phenotype are not yet well characterized. MicroRNAs (miRs) are small noncoding RNAs that play a powerful role in biological processes. Early studies have linked miRs to the control of self-renewal and differentiation in normal and cancer stem cells. We aimed to study the functional role of miRs in human breast cancer stem cells (BCSCs), also named mammospheres. We found that miR-221 was upregulated in BCSCs compared to their differentiated counterpart. Similarly, mammospheres from T47D cells had an increased level of miR-221 compared to differentiated cells. Transfection of miR-221 in T47D cells increased the number of mammospheres and the expression of stem cell markers. Among miR-221s targets, we identified DNMT3b. Furthermore, in BCSCs we found that DNMT3b repressed the expression of various stemness genes, such as Nanog and Oct 3/4, acting on the methylation of their promoters, partially reverting the effect of miR-221 on stemness. We hypothesize that miR-221 contributes to breast cancer tumorigenicity by regulating stemness, at least in part through the control of DNMT3b expression.

miR-340 predicts glioblastoma survival and modulates key cancer hallmarks through down-regulation of NRAS.

Glioblastoma is the most common primary brain tumor in adults; with a survival rate of 12 months from diagnosis. However, a small subgroup of patients, termed long-term survivors (LTS), has a survival rate longer then 12–14 months. There is thus increasing interest in the identification of molecular signatures predicting glioblastoma prognosis and in how to improve the therapeutic approach. Here, we report miR-340 as prognostic tumor-suppressor microRNA for glioblastoma. We analyzed microRNA expression in > 500 glioblastoma patients and found that although miR-340 is strongly down-regulated in glioblastoma overall, it is up-regulated in LTS patients compared to short-term survivors (STS). Indeed, miR-340 expression predicted better prognosis in glioblastoma patients. Coherently, overexpression of miR-340 in glioblastoma cells was found to produce a tumor-suppressive activity. We identified NRAS mRNA as a critical, direct target of miR-340: in fact, miR-340 negatively influenced multiple aspects of glioblastoma tumorigenesis by down-regulating NRAS and downstream AKT and ERK pathways. Thus, we demonstrate that expression of miR-340 in glioblastoma is responsible for a strong tumor-suppressive effect in LTS patients by down-regulating NRAS. miR-340 may thus represent a novel marker for glioblastoma diagnosis and prognosis, and may be developed into a tool to improve treatment of glioblastoma.

MiR-24 induces chemotherapy resistance and hypoxic advantage in breast cancer

Breast cancer remains one of the leading causes of cancer mortality among women. It has been proved that the onset of cancer depends on a very small pool of tumor cells with a phenotype similar to that of normal adult stem cells. Cancer stem cells (CSC) possess self-renewal and multilineage differentiation potential as well as a robust ability to sustain tumorigenesis. Evidence suggests that CSCs contribute to chemotherapy resistance and to survival under hypoxic conditions. Interestingly, hypoxia in turn regulates self-renewal in CSCs and these effects may be primarily mediated by hypoxic inducible factors (HIFs). Recently, microRNAs (miRNAs) have emerged as critical players in the maintenance of pluripotency and self-renewal in normal and cancer stem cells. Here, we demonstrate that miR-24 is upregulated in breast CSCs and that its overexpression increases the number of mammospheres and the expression of stem cell markers. MiR-24 also induces apoptosis resistance through the regulation of BimL expression. Moreover, we identify a new miR-24 target, FIH1, which promotes HIFα degradation: miR-24 increases under hypoxic conditions, causing downregulation of FIH1 and upregulation of HIF1α. In conclusion, miR-24 hampers chemotherapy-induced apoptosis in breast CSCs and increases cell resistance to hypoxic conditions through an FIH1−HIFα pathway.

Aptamer-miR-34c conjugate affects cell proliferation of non-small cell lung cancer cells

MicroRNAs (miRNAs) are key regulators of different human processes that represent a new promising class of cancer therapeutics or therapeutic targets. Indeed, in several tumor types, including non-small-cell lung carcinoma (NSCLC), the deregulated expression of specific miRNAs has been implicated in cell malignancy. As expression levels of the oncosuppressor miR-34c-3p are decreased in NSCLC compared to normal lung, we show that reintroduction of miR-34c-3p reduces NSCLC cell survival in vitro. Further, in order to deliver the miR-34c-based therapeutic selectively to tumor cells, we took advantage of a reported nucleic acid aptamer (GL21.T) that binds and inhibits the AXL transmembrane receptor and is rapidly internalized in the target cells. By applying methods successfully used in our laboratory, we conjugated miR-34c to the GL21.T aptamer as targeting moiety for the selective delivery to AXL-expressing NSCLC cells. We demonstrate that miR-34c-3p and the GL21.T/miR-34c chimera affect NSCLC cell proliferation and are able to overcome acquired RTK-inhibitor resistance by targeting AXL receptor. Thus, the GL21.T/miR-34c chimera exerts dual inhibition of AXL at functional and transcriptional levels and represents a novel therapeutic tool for the treatment of NSCLC.

La «cura» del patrimonio costruito di uso pubblico. istituzioni, regolamenti e prassi a Napoli tra Cinque e Settecento

Tra le piu antiche istituzioni napoletane, il Tribunale della Fortificazione (poi Mattonata e Acqua) ha controllato in ambito urbano l’insieme delle pratiche connesse alla manutenzione delle mura, di bastioni e torri, delle attrezzature idrauliche - acquedotti e fontane - nonche degli spazi aperti di giurisdizione pubblica dal Medioevo agli inizi del diciannovesimo secolo. L’ampia documentazione archivistica riferita all’organizzazione amministrativa, agli interventi sollecitati o imposti e, ancora, alle numerose figure di ingegneri e tavolari coinvolti tra la prima meta del Cinquecento e il 1805 nella progettazione e direzione delle opere, conservata nell’Archivio storico municipale di Napoli, e indagata nello scritto con riferimento al periodo compreso tra la meta del Cinquecento e il primo Settecento. Il puntuale controllo sulle condizioni delle strade e piazze, la costante pratica della manutenzione degli acquedotti, delle fontane cittadine, del corso delle acque piovane e delle pavimentazioni stradali come il mantenimento degli spazi pubblici liberi da costruzioni private trovano ampia dimostrazione nella ricca documentazione conservata e risparmiata dall’incendio che nel 1946 ha distrutto parte del corpus archivistico. Attraverso lo studio dei Banni, Appuntamenti e Conclusioni e analizzato in modo sistematico un tema di ricerca ancora non sufficientemente approfondito in letteratura, riguardante il ruolo e gli obiettivi che il governo vicereale attribuiva alla cura del patrimonio urbano collettivo nonche alle modalita tecniche con cui quest’ultima e condotta in eta barocca.

Torres costeras durante el siglo XVI. Estrategias territoriales y técnicas constructivas en el frente marítimo levantino del Reino de Aragón y Virreinato de Nápoles

The historical strong relationship between the eastern coast of Spain and southern part of Italian Peninsula has been a common issue over different civilizations from Phoenicians, Greeks and Romans up to the most recent modern kingdoms. Considering this set of connections based on trade routes and territorial bilateral interests, the western shores of Mare Nostrum have been key-points of a dense network of military and economical strategies. The dynamics of extensive territorial control have left interesting examples of costal watchtowers both in the Sorrento Peninsula, near Naples (Italy) and in the East coast of Valencia (Spain). The process of towers construction begins at early stages of Middle Age and, from the 16th century it runs up into a systemic control of the coastline, thanks to the reuse of obsolete fortified elements and new building-site procedures. In the paper, the authors aim at highlighting the territorial relationships among a number of towers built during the 16th century, as well as they propose a comparison of their construction techniques in order to gain an historical comparative and a contextual understanding of these fortified systems. DOI: http://dx.doi.org/10.4995/FORTMED2015.2015.1689

Abstract 3126: Survival in glioblastoma cancer patients is predicted by miR-340, that regulates key cancer hallmarks by inhibiting NRAS

Glioblastoma is the most frequent brain tumor in adults and is the most lethal form of human cancer. Despite the improvements in treatments, patients survival remains poor. Nevertheless, a subset of patients survives longer than 3 years and they are classified as long-term survivors (LTS). The molecular and cellular mechanisms underlying the rare phenomenon of LTS are not well known. MicroRNAs (miRNAs or miRs) are a class of endogenous non-coding RNA of 19-24 nucleotides in length that have an important role in the negative regulation of gene expression. Deregulation of miRNAs is linked to the pathogenesis of many types of cancer. We performed a nanostring miRNA analysis on long and short glioblastoma survivors patients to individuate miRs potentially involved in LTS phenotype. Using this approach, we identified miR-340 as a novel tumor-suppressor-miR in glioblastoma. miR-340 expression was increased in LTS compared to short term survivors. Moreover, a log-rank test analysis on both the data collected from TCGA database and from a cohort of glioblastoma patients present in our lab revealed that low levels of miR-340 were a risk indicator for glioblastoma patients survival. miR-340 overexpression decreased cell proliferation, anchorage independent cell growth, cell cycle and response to temozolomide in different glioblastoma cell lines. We identified NRAS as a direct target of miR-340. Rescue experiments showed its essential role in mediating the onco-suppressive activity of miR-340. The overexpression of miR-340 decreased the activation of ERKs and AKT, the main pathways downstream NRAS. Furthermore, glioblastoma cells stably infected with a lentivirus encoding miR-340 exhibited a drastic reduction of tumor growth in nude mice. In conclusion, our findings reveal miR-340 as a new tumor-suppressor miRNA up-regulated in LTS in glioblastoma. Its expression inversely correlates with survival of glioblastoma patients. miR-340 is able to regulate multiple tumorigenic features of glioblastoma cells, offering a novel potential prognostic and therapeutical target for glioblastoma. Citation Format: Danilo Fiore, Cristina Quintavalle, Elvira Donnarumma, Giuseppina Roscigno, Margherita Iaboni, Valentina Russo, Assunta Adamo, Fabio De Martino, Adelaide Greco, Giulia Romano, Soini Ylermi, Arturo Brunetti, Carlo Maria Croce, Gerolama Condorelli. Survival in glioblastoma cancer patients is predicted by miR-340, that regulates key cancer hallmarks by inhibiting NRAS. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3126. doi:10.1158/1538-7445.AM2015-3126

Interdisciplinary Research about Bourbon's Architectural Heritage: the Case-study of the Immacolatella in Naples

The Immacolatella building in the port of Naples is one of the last examples of the modern planning supported by Charles III in order to make Naples the new capital of the Spanish Reign of Bourbon in mid-18th century. It is a very important symbol in the Neapolitan memory of the emigration to the United States in early 1900s. Despite its historical value, the building is affected by severe decay due to both negligence and weathering, which are caused by its onshore location. This paper shows the results of interdisciplinary research undertaken in the University of Naples with the aim of giving a preliminary knowledge for its preservation. By means of historical sources and mensio-chronological analyses, the building development was retraced. Moreover, laboratory tests were carried out in order to collect information about geomaterials. The collected data show the high vulnerabilities of this Royal building and the need to create a multidisciplinary strategy for its preservation.