Piano I.,G B Bietti Foundation For Ophthalmology |
Novelli E.,G B Bietti Foundation For Ophthalmology |
Gasco P.,Nanovector srl |
Ghidoni R.,University of Milan |
And 2 more authors.
European Journal of Neuroscience | Year: 2013
The prevention of cone loss during retinal degeneration is a major goal of most therapeutic strategies in retinal degenerative diseases. An intriguing issue in the current research in this field is to understand why a genetic mutation that affects rods eventually leads to cone death. The main objective of the present study was to investigate to what extent rescuing rods from degeneration affects the survival of cones and prevents functional impairment of the visual performance. To this purpose, we compared rod and cone viabilities by both ex vivo and in vivo determinations in the rd10 mutant mouse, a validated model of human retinitis pigmentosa. The ex vivo experiments included morphological and biochemical tests, whereas in vivo studies compared the rod-mediated scotopic with the cone-mediated photopic electroretinogram. We also determined the overall visual performance by behaviorally testing the visual acuity (VA). The electroretinogram measurements showed that the kinetics of the photopic response in rd10 mice was slowed down with respect to the age-paired wild-type at a very early stage of the disease, when rods were still present and responsive. We then tested cone viability and function under a pharmacological scheme previously shown to prolong rod survival. The treatment consisted of eye drop administration of myriocin, an inhibitor of the biosynthesis of ceramide, a powerful proapoptotic messenger. The results of biochemical, morphological and functional assays converged to show that, in treated rd10 mice cone photoreceptors, the inner retina and overall visual performance were preserved well after rod death. © 2013 Federation of European Neuroscience Societies and Blackwell Publishing Ltd.
Minelli R.,University of Turin |
Occhipinti S.,University of Turin |
Gigliotti C.L.,The Interdisciplinary Center |
Barrera G.,University of Turin |
And 8 more authors.
British Journal of Pharmacology | Year: 2013
Background and Purpose Solid lipid nanoparticles containing cholesteryl butyrate (cholbut SLN) can be a delivery system for the anti-cancer drug butyrate. These nanoparticles inhibit adhesion of polymorphonuclear and tumour cells to endothelial cells and migration of tumour cells, suggesting that they may act as anti-inflammatory and anti-tumour agents. Here we have evaluated the effects of cholbut SLN on tumour cell growth using in vitro and in vivo models. Experimental Approach Cholbut SLNs were incubated with cultures of four tumour cell lines, and cell growth was analysed by assessing viability, clonogenic capacity and cell cycle. Effects on intracellular signalling was assessed by Western blot analysis of Akt expression. The in vivo anti-tumour activity was measured in two models of PC-3 cell xenografts in SCID/Beige mice. Key Results Cholbut SLN inhibited tumour cell line viability, clonogenic activity, Akt phosphorylation and cell cycle progression. In mice injected i.v. with PC3-Luc cells and treated with cholbut SLN,. in vivo optical imaging and histological analysis showed no metastases in the lungs of the treated mice. In another set of mice injected s.c. with PC-3 cells and treated with cholbut SLN when the tumour diameter reached 2 mm, analysis of the tumour dimensions showed that treatment with cholbut SLN substantially delayed tumour growth. Conclusion and Implications Cholbut SLN were effective in inhibiting tumour growth in vitro and in vivo. These effects may involve, in part, inhibition of Akt phosphorylation, which adds another mechanism to the activity of this multipotent drug. © 2013 The British Pharmacological Society.
Agency: European Commission | Branch: FP7 | Program: CP-FP | Phase: NMP-2008-1.1-1 | Award Amount: 4.86M | Year: 2009
The aim of this project is the assembly and the fabrication of a new generation of multifunctional nanostructures for performing combined hyperthermia and controlled drug release, specifically targeted to cancer cells. The magnetic nanocontainers we intend to develop can perform at the same time cell recognition, hyperthermia treatment, and, as a consequence of the heat and /or cell environment stimuli, the release of drug with high selectivity for ovarian carcinoma. These multifunctional tasks are made possible due to the inclusion of three main components: a) the magnetic nanoparticles, allowing detection by MRI, cancer treatment by hyperthermia and providing stimuli for drug release; b) the nanocontainers, which allow for drug encapsulation and protection from degradation, facilitate the release of the drug upon application of an external stimulus, such as heat, or an internal one, such as the acidic pH of the tumour cells; c) the antibody fragments attached to the surface of the magnetic nanocontainers to deliver them selectively to the ovarian cancer cells. The individual building blocks and their assemblies will be characterized with respect to physical, chemical, and biological features, followed by dissemination of the newly acquired knowledge. Cell culture experiments will allow to understand the performance of such nano-tools in vitro. Directed towards application in patients, in vivo animal studies will be carried out on the most successful magnetic nanocontainers. The objectives of this proposal cover a wide range of scientific fields, hence a truly interdisciplinary collaboration between chemists, physicists, and biologists is required. To this end, we propose a european network collaboration between academic partners, who will take care of the development of new solutions for nanofabrication, and industrial partners implied in the field of the proposed application who will evaluate/develop the materials and act as advisors for risks arising during the project.
Agency: European Commission | Branch: FP7 | Program: CP-FP | Phase: NMP.2012.1.4-2 | Award Amount: 5.18M | Year: 2013
UNION will develop nanoparticle (NP) assembly techniques, and assembly monitoring technologies to prepare novel hierarchically-ordered nanoparticle clusters (NPCs). By improving control over the synthesis and assembly of NPs we will produce materials with tailored and predictable properties. Furthermore, by incorporating hierarchical control into the assembly (through the type, size and spatial distribution of the NPs) it will be possible to assess the influence of the hierarchy on properties and develop new functionalities. UNION will investigate how the emergent properties of the assemblies are determined by the architecture of the assembly, the extent of order, and the properties of the component NPs. This will enable tuning of the primary NP properties and the assembly processes to develop significant breakthroughs in nano-devices and next generation complex nanotechnology products. As the ultimate aim is commercial exploitation of our results, in each stage of the development process we will use application driven, scalable and cost-effective processes, incorporating EHS assessment and roadmap preparation towards future industrial deployment. UNION will achieve its objectives through a three stage approach. - Improved NP preparation providing optimised surface chemistry for subsequent assembly - Novel NPC formation (hierarchical nanoparticle assembly) methods - Roll-out of NPCs for three application areas NPC applications will be developed within three core areas corresponding to the different hierarchical structural levels; in suspensions of individual NPCs (biomedical), in supported 2D NPC arrays (optical), and in 3D arrays or nanocomposites (thermoelectric). Our consortium is comprised of multidisciplinary research groups involving 8 partners with ex-pertise in preparation and application of nano-materials. It includes significant industrial participation with 4 companies with specific knowledge and testing capability for the target application areas.
Agency: European Commission | Branch: FP7 | Program: CP-IP | Phase: NMP-2007-4.0-4 | Award Amount: 14.37M | Year: 2008
The search for effective therapies and early detection strategies for Alzheimers Disease (AD), the major cause of dementia in Europe, is imperative. It is known that -amyloid (A) peptide plays a central role in neurodegeneration. In AD brain, A is released in a soluble form that progressively becomes insoluble forming aggregates; extracellular plaques mainly composed of A are a hallmark of post-mortem brains. These premises strongly suggest brain A as a possible target for therapy and diagnosis of AD. In addition, it is known that brain and blood A pools are in equilibrium via the blood-brain-barrier (BBB). Accordingly, it has been reported that removal of blood A may withdraw the excess of brain A by a sink effect. Thus, blood A is another potential target. The aim of this project is to utilize nanoparticles (NPs) specifically engineered for targeting brain A, for the combined diagnosis and therapy (theranostics) of AD. NPs (liposomes, solid lipid NPs, polymeric-NPs) will be multiple-functionalized with: i) a large arsenal of molecules (specific lipids, antiamyloidogenic drugs, polyphenols, heteroaromatic compounds, unnatural peptides and peptidomimetics, antibodies) interacting with A in all aggregation forms, ii) PET or MRI contrast agents detecting such interaction, iii) molecules stimulating BBB crossing via the transcytotic route. Several artificial and cellular models will be used to fine-tune such features and to improve NPs biocompatibility, non-immunogenicity, non-toxicity and physical stability. Eventually, absorption, distribution, metabolism and excretion will be studied using animal models of AD. Different routes (i.v., oral, nasal) and protocols (two-step, NPs cocktails, aerosols) of administration will be utilized to boost NPs brain delivery. The prediction is that NPs will detect, disaggregate and remove A brain deposits. In any case, NPs will interact with blood A, withdrawing the excess of brain peptide by a sink effect.
Caretti A.,University of Milan |
Bragonzi A.,San Raffaele Scientific Institute |
Facchini M.,San Raffaele Scientific Institute |
De Fino I.,San Raffaele Scientific Institute |
And 7 more authors.
Biochimica et Biophysica Acta - General Subjects | Year: 2014
Background: Sphingolipids take part in immune response and can initiate and/or sustain inflammation. Various inflammatory diseases have been associated with increased ceramide content, and pharmacological reduction of ceramide diminishes inflammation damage in vivo. Inflammation and susceptibility to microbial infection are two elements in a vicious circle. Recently, sphingolipid metabolism inhibitors were used to reduce infection. Cystic fibrosis (CF) is characterized by a hyper-inflammation and an excessive innate immune response, which fails to evolve into adaptive immunity and to eradicate infection. Chronic infections result in lung damage and patient morbidity. Notably, ceramide content in mucosa airways is higher in CF mouse models and in patients than in control mice or healthy subjects. Methods: The therapeutic potential of myriocin, an inhibitor of the sphingolipid de novo synthesis rate limiting enzyme (Serine Palmitoyl Transferase, SPT),was investigated in CF cells and mice models. Results We treated CF human respiratory epithelial cells with myriocin, This treatment resulted in reduced basal, as well as TNFα-stimulated, inflammation. In turn, TNFα induced an increase in SPT in these cells, linking de novo synthesis of ceramide to inflammation. Furthermore, myriocin-loaded nanocarrier, injected intratrachea prior to P. aeruginosa challenge, enabled a significant reduction of lung infection and reduced inflammation. Conclusions: The presented data suggest that de novo ceramide synthesis is constitutively enhanced in CF mucosa and that it can be envisaged as pharmacological target for modulating inflammation and restoring effective innate immunity against acute infection. General significance Myriocin stands as a powerful immunomodulatory agent for inflammatory and infectious diseases. © 2013 Elsevier B.V. All rights reserved.
Agency: European Commission | Branch: FP7 | Program: MC-IAPP | Phase: FP7-PEOPLE-2013-IAPP | Award Amount: 2.37M | Year: 2013
Antimicrobial agents, such as antibiotics, have dramatically reduced the number of deaths from infectious diseases over the last 70 years. However, through overuse and misuse of these agents, many micro-organisms have developed antimicrobial resistance. Oligonucleotide therapeutics have the potential to become the new class of antibacterials capable of treating a broad range of infections. By acting on novel targets, they circumvent current resistance mechanisms and with judicious use, can suppress the rise of future resistance. DNA-TRAP will build on a platform technology that uses proprietary nucleic acid-based Transcription Factor Decoys (TFDs) that act on novel genomic targets by capturing key regulatory proteins to block essential bacterial genes and defeat infection. Taking forward newly emerging insights and expertise that exists within each of the partners and through the mutual secondment of researchers, the project aims to develop a new class of nanoparticulate antibacterials capable of meeting the clinical challenge of drug-resistant infections such as Clostridium difficile and Pseudomonas aeruginosa. DNA-TRAP will establish a lasting, international partnership for transfer of knowledge between Industry and Academia in the field of nanomedicine. Exchange of knowledge and expertise between the partners is key to establishing the fundamental properties of nanostructured drug delivery systems to treat bacterial infections and through this, provide the basis for building a manufacturing platform to advance the experimental therapeutic into clinical trials. 17 researchers in the field of drug development and delivery from 2 commercial (SME) and 2 non-commercial partners across 2 member states, will have the opportunity to share and acquire new complementary and multidisciplinary knowledge, through inter-sectoral and interdisciplinary exchange, allowing for the development of new solutions and the establishment of further joint research projects.
University of Milan, Nanovector S.R.L., University of Pisa and National Research Council Italy | Date: 2010-02-23
The method for preventing and delaying inherited retinal degenerations using serine palmitoyltransferase inhibitors, and compositions which contain them.
Gobbi M.,Mario Negri Institute for Pharmacological Research |
Re F.,University of Milan Bicocca |
Canovi M.,Mario Negri Institute for Pharmacological Research |
Beeg M.,Mario Negri Institute for Pharmacological Research |
And 8 more authors.
Biomaterials | Year: 2010
The neurotoxic beta-amyloid peptide (Aβ), formed in anomalous amounts in Alzheimer's disease (AD), is released as monomer and then undergoes aggregation forming oligomers, fibrils and plaques in diseased brains. Aβ aggregates are considered as possible targets for therapy and/or diagnosis of AD. Since nanoparticles (NPs) are promising vehicles for imaging probes and therapeutic agents, we realized and characterized two types of NPs (liposomes and solid lipid nanoparticles, 145 and 76 nm average size, respectively) functionalized to target Aβ1-42 with high affinity. Preliminary immunostaining studies identified anionic phospholipids [phosphatidic acid (PA) and cardiolipin (CL)] as suitable Aβ1-42 ligands. PA/CL-functionalized, but not plain, NPs interacted with Aβ1-42 aggregates as indicated by ultracentrifugation experiments, in which binding reaction occurred in solution, and by Surface Plasmon Resonance (SPR) experiments, in which NPs flowed onto immobilized Aβ1-42. All these experiments were carried out in buffered saline. SPR studies indicated that, when exposed on NPs surface, PA/CL display very high affinity for Aβ1-42 fibrils (22-60 nm), likely because of the occurrence of multivalent interactions which markedly decrease the dissociation of PA/CL NPs from Aβ Noteworthy, PA/CL NPs did not bind to bovine serum albumin. The PA/CL NPs described in this work are endowed with the highest affinity for Aβ so far reported. These characteristics make our NPs a very promising vector for the targeted delivery of potential new diagnostic and therapeutic molecules to be tested in appropriate animal models. © 2010 Elsevier Ltd.
PubMed | University of Lausanne, CSIC - Institute of Advanced Chemistry of Catalonia, Nanovector Srl. and University of Milan
Type: Journal Article | Journal: Basic research in cardiology | Year: 2016
The injury caused by myocardial reperfusion after ischemia can be contained by interventions aimed at reducing the inflammation and the oxidative stress that underlie exacerbation of tissue damage. Sphingolipids are a class of structural and signaling lipid molecules; among them, the inflammation mediator ceramide accumulates in the myocardium upon ischemia/reperfusion. Here, we show that, after transient coronary occlusion in mice, an increased de novo ceramide synthesis takes place at reperfusion in the ischemic area surrounding necrosis (area at risk). This correlates with the enhanced expression of the first and rate-limiting enzyme of the de novo pathway, serine palmitoyltransferase (SPT). The intraventricular administration at reperfusion of myriocin, an inhibitor of SPT, significantly protected the area at risk from damage, reducing the infarcted area by 40.9 % relative to controls not treated with the drug. In the area at risk, myriocin downregulated ceramide, reduced the content in other mediators of inflammation and reactive oxygen species, and activated the Nrf2-HO1 cytoprotective response. We conclude that an enhanced ceramide synthesis takes part in ischemia/reperfusion injury and that myriocin treatment can be proposed as a strategy for myocardial pharmacological postconditioning.