Stroma-derived miR-214 coordinates tumor dissemination

BackgroundTumor progression is based on a close interaction between cancer cells and Tumor MicroEnvironment (TME). Here, we focus on the role that Cancer Associated Fibroblasts (CAFs), Mesenchymal Stem Cells (MSCs) and microRNAs (miRs) play in breast cancer and melanoma malignancy.MethodsWe used public databases to investigate miR-214 expression in the stroma compartment of primary human samples and evaluated tumor formation and dissemination following tumor cell injections in miR-214 overexpressing (miR-214(over)) and knock out (miR-214(ko)) mice. In addition, we dissected the impact of Conditioned Medium (CM) or Extracellular Vesicles (EVs) derived from miR-214-rich or depleted stroma cells on cell metastatic traits.ResultsWe evidence that the expression of miR-214 in human cancer or metastasis samples mostly correlates with stroma components and, in particular, with CAFs and MSCs. We present data revealing that the injection of tumor cells in miR-214(over) mice leads to increased extravasation and metastasis formation. In line, treatment of cancer cells with CM or EVs derived from miR-214-enriched stroma cells potentiate cancer cell migration/invasion in vitro. Conversely, dissemination from tumors grown in miR-214(ko) mice is impaired and metastatic traits significantly decreased when CM or EVs from miR-214-depleted stroma cells are used to treat cells in culture. Instead, extravasation and metastasis formation are fully re-established when miR-214(ko) mice are pretreated with miR-214-rich EVs of stroma origin. Mechanistically, we also show that tumor cells are able to induce miR-214 production in stroma cells, following the activation of IL-6/STAT3 signaling, which is then released via EVs subsequently up-taken by cancer cells. Here, a miR-214-dependent pro-metastatic program becomes activated.ConclusionsOur findings highlight the relevance of stroma-derived miR-214 and its release in EVs for tumor dissemination, which paves the way for miR-214-based therapeutic interventions targeting not only tumor cells but also the TME

RNA delivery by extracellular vesicles in mammalian cells and its applications.

The term 'extracellular vesicles' refers to a heterogeneous population of vesicular bodies of cellular origin that derive either from the endosomal compartment (exosomes) or as a result of shedding from the plasma membrane (microvesicles, oncosomes and apoptotic bodies). Extracellular vesicles carry a variety of cargo, including RNAs, proteins, lipids and DNA, which can be taken up by other cells, both in the direct vicinity of the source cell and at distant sites in the body via biofluids, and elicit a variety of phenotypic responses. Owing to their unique biology and roles in cell-cell communication, extracellular vesicles have attracted strong interest, which is further enhanced by their potential clinical utility. Because extracellular vesicles derive their cargo from the contents of the cells that produce them, they are attractive sources of biomarkers for a variety of diseases. Furthermore, studies demonstrating phenotypic effects of specific extracellular vesicle-associated cargo on target cells have stoked interest in extracellular vesicles as therapeutic vehicles. There is particularly strong evidence that the RNA cargo of extracellular vesicles can alter recipient cell gene expression and function. During the past decade, extracellular vesicles and their RNA cargo have become better defined, but many aspects of extracellular vesicle biology remain to be elucidated. These include selective cargo loading resulting in substantial differences between the composition of extracellular vesicles and source cells; heterogeneity in extracellular vesicle size and composition; and undefined mechanisms for the uptake of extracellular vesicles into recipient cells and the fates of their cargo. Further progress in unravelling the basic mechanisms of extracellular vesicle biogenesis, transport, and cargo delivery and function is needed for successful clinical implementation. This Review focuses on the current state of knowledge pertaining to packaging, transport and function of RNAs in extracellular vesicles and outlines the progress made thus far towards their clinical applications

Endocrine disruptors and nuclear receptors in the control of neural progenitors proliferation

Endocrine-disrupting chemicals (EDCs), in particular plasticizers present in food wrapping materials, are very relevant to human health. Several studies have shown that EDCs may pose the greatest risk during prenatal and early postnatal development, especially when binding to members of the nuclear receptor (NR) superfamily. EDCs can mimic/suppress estrogen actions in the developing brain by binding to estrogen receptors (ERs), and also interfere with neural progenitors (NPs) homeostasis through activation of peroxisome proliferator-activated receptors (PPARs) and retinoid X receptors (RXRs). We used the ST14A immortalized neural progenitor cell line to simulate the effects of prenatal NPs exposure to EDCs. We found by RT-PCR that the following NRs were expressed: ERα, ERβ, PPARα, PPARβ, PPARγ, RXR. Plasticizers were chosen based on their computational affinity for these receptors. NPs were exposed for 24-48 hours to endogenous and synthetic estrogens (17-β-estradiol and ethinyl estradiol) and to the following plasticizers: Bisphenol A, Diisononyl phthalate, Diisodecyl phthalate, Diethileneglicol benzoate. Both estrogens were able to increase cell proliferation by about 30% at 24hrs. Bisphenol-A (10nM) had an estrogen-like behavior in enhancing cell number, but its action was still significant at 48hrs. At 200nM concentration, all plasticizers determined an early increase in proliferation, however only DiDP and DGB were able to sustain this positive modulation for 48hrs. Ongoing studies in our lab will determine the cellular pathways activated by these EDCs in NPs, in order to earn greater insight into how these molecules may influence and perturb the neurogenic process

Combined in silico and in vitro studies of phthalates and organophosphorus compounds: effects on peroxisome proliferator activated receptors (PPARs) signalling pathways

Increasing evidence suggests that some environmental contaminants, including known endocrine disrupting chemicals (EDCs), are able to interfere with metabolic pathways by interacting with nuclear receptors. Recently, it has been demonstrated that phthalates and organometallic compounds bind to peroxisome proliferator activated receptors (PPARs) resulting in modulation of lipid metabolism at both the systemic and peripheral level. However, little is known about the metabolic impact of these pollutants on fish. In this context, we performed an in silico docking screen of an EDCs database to identify a set of ligands with conveniently high affinity for the PPARs. Kd values in the nanomolar to micromolar range, generated by the in silico model, suggest that piscine PPARs may be activated by phthalates (e.g. di-isononylphpthalate -DiNP; Di-isodecyl phthalate-DiDP) and some organophosphorus compounds (e.g. tri-m-cresyl phosphate-TmCP) at concentrations similar to those activating the homologous mammalian receptors. Because natural endogenous ligands for PPARs are involved in lipid homeostasis, we assessed the effects of compounds identified using in silico screening on Sparus aurata hepatocytes primary cultures. Generally, exposure of hepatocytes to 0.1, 1 or 10 microM of DiNP, DiDP or TmCP consistently increased both PPAR and its heterodimeric partner Retinoid X Receptor (RXR) mRNA levels at 48 h. In addition, all compounds investigated produced significant increases in the expression of the PPAR target genes, carnitine palmitoyltransferase (CPT) isoforms. In general our data show that phthalates and TmCP modulated PPAR signaling in the seabream in vitro system. The results also suggest the potential involvement of these pollutants in the modulation of mitochondrial fatty acid oxidation

Il Il problema della mandibola nella chirurgia dei tumori tonsillari: proposta di una tecnica.

The Authors report a technique of a conservative transmandibular approach for the surgical treatment of tonsillar region malignancies. The procedure is based upon the preparation of a vascular mandibular flap through two osteotomies, which are performed, the first above the mandibular foramen and the second below the mental foramen. The main advantages of present technique are: lip-chin-splitting is avoided and, due to the preservation of mandibular vascular and nervous supply, it is possible to accomplish, without risk, postoperative radiotherapy. Moreover, if dictated by oncologic reasons, this conservative procedure may easily be transformed into a radical surgery (commando or neck-jaw operation)

Endocrine disruptors present in food packages do act as lipid metabolism modulators.

Endocrine-disrupting chemicals (EDCs) may cause several noxious problems to both humans and wildlife and their modulations on energy balance and lipid metabolism are nowadays of great concern. Among environmental EDCs, plasticizers employed in food wrapping materials are very relevant due to their widespread distribution and persistence in the food chain. In the present work we first identify by in silico screening compounds, among the permitted plasticizers for food packaging (EU Regulation, 10/2011), with high predicted affinity for nuclear receptors known to mediate obesogenic effects, such as PPARs, LXR and RXR. The following molecules were therefore chosen for the subsequent studies: the well-known, bisphenol-A, three phthalates (di-isononylphpthalate, DiNP; Di-isodecyl phthalate, DiDP; diethylene glycol dibenzoate, DiGD) and one organophosphorus (tri-m-cresyl phosphate,TmCP).The predicted affinity for the nuclear receptors was then verified in vitro with a luciferase assay: DiNP, DiDP and DiGD were shown to have activity on PPAR α/γ at 10-5-10-7M and to induce the expression of downstream target genes (FABP4, PDK4, FGF21, CPT2). The potential obesogenic effects of the selected compounds were then tested on 3T3-L1 murine preadipocytes cells, a widely used model for adipogenesis in vitro. All the molecules, when used in a range of 10-8-10-6M, were able to significantly enhance lipid droplets deposition, both when administered during the two-days differentiation induction, as well as when maintained for all the eight-days post-differentiation period. Food plastic packages are mostly composed by a mixture of plasticizers so that many molecules at the same time may migrate and contaminate food. We consequently treated 3T3-L1 cells with mixtures of the selected plasticizers (10-8M each), coherently with some commercial plastics (pvc, polypropylene, polyethylene teraphthalate), and we observed in all the cases a significant positive modulation on lipid deposition. The obesogenic effects of the compounds were also investigated by performing qRT-PCR on target genes such as PPARs, LXR, RXR and their downstream effectors. Taken together, our data enforce the emerging awareness on energy balance and lipid metabolism modifications following environmental exposure that could be of concern in vulnerable periods, such as early postnatal life

Endocannabinoids are involved in male vertebrate reproduction: regulatory mechanisms at central and gonadal level

Endocannabinoids (eCBs) are natural lipids regulating a large array of physiological functions and behaviors in vertebrates. The eCB system is highly conserved in evolution and comprises several specific receptors (type-1 and type-2 cannabinoid receptors), their endogenous ligands (e.g., anandamide and 2-arachidonoylglycerol), and a number of biosynthetic and degradative enzymes. In the last few years, eCBs have been described as critical signals in the control of male and female reproduction at multiple levels: centrally, by targeting hypothalamic gonadotropin-releasing-hormone-secreting neurons and pituitary, and locally, with direct effects on the gonads. These functions are supported by the extensive localization of cannabinoid receptors and eCB metabolic enzymes at different levels of the hypothalamic–pituitary–gonadal axis in mammals, as well as bonyfish and amphibians. In vivo and in vitro studies indicate that eCBs centrally regulate gonadal functions by modulating the gonadotropin-releasing hormone–gonadotropin–steroid network through direct and indirect mechanisms. Several proofs of local eCB regulation have been found in the testis and male genital tracts, since eCBs control Sertoli and Leydig cells activity, germ cell progression, as well as the acquisition of sperm functions. A comparative approach usually is a key step in the study of physiological events leading to the building of a general model.Thus, in this review, we summarize the action of eCBs at different levels of the male reproductive axis, with special emphasis, where appropriate, on data from non-mammalian vertebrates

Plasticizers present in food packaging significantly affect lipid metabolism

Recent data suggest that endocrine-disrupting chemicals (EDCs) may alter lipid metabolism and energy balance. Among environmental EDCs, plasticizers employed in food packaging are of great concern due to their widespread distribution and high persistency in the food chain. We first selected by in silico screening several compounds from a list of plasticizers approved for food packaging (EU Regulation 10/2011) showing high affinity for lipogenic nuclear receptors (PPARs, LXR and RXR). The following plasticizers were chosen: the well-known bisphenol-A, three phthalates (diisononylphpthalate, DiNP; Di-isodecyl phthalate, DiDP; diethylene glycol dibenzoate, DiGD) and one organophosphorus compound (tri-m-cresylphosphate,TmCP).Their binding capacity to nuclear receptors was verified on HEPG2 cells transfected with receptor-luciferase reporter constructs; all phtalates (10-5-10-7 M) were shown to activate PPAR and to induce the expression of downstream target genes (FABP4, PDK4, FGF21, CPT2). The potential obesogenic effects of these compounds were then tested on differentiating 3T3-L1 preadipocyte cells. All molecules (10-8-10-6 M) were able to significantly enhance lipid droplet deposition when administered both in the two-days differentiation induction, or in the eight-days post differentiation period. Food plastic packaging is mostly composed by mixtures of plasticizers, so multiple molecules may migrate simultaneously into food. We therefore treated 3T3-L1 cells with mixtures of plasticizers (10-8 M each), mimicking commercial plastics (pvc, polypropylene, polyethylene teraphthalate); in all cases we observed a significant positive modulation of lipid deposition. The obesogenic effects of all compounds were also investigated at the molecular level by performing qRT-PCR on target genes such as PPARs, LXR, RXR and their downstream effectors. Taken together, our data enforce the emerging awareness on alteration of lipid metabolism following environmental exposure to plasticizers. This effect could be of special concern in vulnerable periods such as pregnancy and early postnatal life