Project #280

Project #280 - OcuPharm Diagnostics SL - Spain

Date: 2007/04/23 / Deadline: 2010/02/28

Contact

Organisation / OcuPharm Diagnostics SL / Department / Administration
Contact person / Colligris, Mr. Basilio
Email /
Address / Arcos De Jalon sn
Postcode / 28037 / City / Madrid
Country / Spain
Telephone / +34913946859 / Fax / +34913946885
Website /

Familiar with the European Framework Programme? YES

PROJECT

Title: APPROACHING THE TREATMENT of CHONDRODYSPLASIAS by MEANS OF SHORT INTERFERING RNAs / Acronym: BES
Project type / Large-scale integrating collaborative project, Research for the benefit of SMEs
Status / Planned for submission
Call references / Call 2nd
Priorities’ Main Research Areas / This project should aim to exploit emerging gene therapy tools and technologies, such as use of genome editing and repair (RNAi, site specific recombination, etc.) to correct genetic defects, or new gene transfer techniques (novel virus vectors, targeted nanoparticles, bacterial DNA-based vector, bactofection, etc.), which
overcome the limitations of existing tools and for which preclinical proof of concept has already been established.
Workprogramme Topic (according to each priority workprogramme) / Gene therapy - HEALTH-2007-1.4-4: Development of emerging gene therapy tools and technologies for clinical application.
Project description
Chondrodysplasias are a group of pathologies characterized by different mutation in the gene that codifies for the fibroblast growth factor receptor type 3 (FGFR3) in cartilage cells, the chondrocytes, which produce the pathology known as dwarfism. As a consequence of such mutations the FGFR3 receptor is over-activated producing important changes at the intracellular level.
Among them there is an increase in the rate of maturation which leads these cells to apoptosis, changing the biochemical behaviour of these cells as well as the extracellular matrix they produce. The consequence of these processes is a quick ossification which impedes the bone to develop in length properly. Our aim is to study in depth the pathophysiology of chondrodysplasias by means of proteomics and also to investigate the involvement of miRNA in this group of pathologies. Moreover we wish to reduce the increased activity of the mutated FGFR3 receptor by means of small interfering RNA (siRNA) in order to allow a normal bone development.
As a complement to the classical proteomic approaches, investigating the second messenger cascades, we will also study the role of another group of small RNAs (miRNAs) in the natural course of chondrodysplasia. MiRNAs are very similar to siRNAs but are encoded by the genome and are expressed naturally. They ensure proper development by regulating gene expression through the same pathway as siRNAs. At least one miRNA (miR-140) is specifically expressed in cartilage tissue and we have shown that the expression of histone deacetylase 4 (HDAC4) is regulated by miR-140.Since HDCA4 knock-out mice display premature ossification of developing bones due to ectopic and early onset of chondrocyte hypertrophy that is similar to chondrodysplasia, increased expression of miR-140 could lead to chondrodysplasia. We will test whether suppression of miR-140 by antagonists is an alternative approach to ameliorate the symptoms of chondrodysplasia using both in vitro and in vivo approaches. We will also investigate the expression level of all miRNAs using miRNA micro arrays in normal chondrocyte cells and cell lines carrying the mutant FGFR3 gene and also in cartilage tissues from wild type and achondroplasic mice to find out whether any miRNA shows altered accumulation. If any miRNA shows increased or decreased accumulation in the diseased condition, we will test whether targeting or expressing them, respectively, can affect the symptoms.
Furthermore during the project we will use the siRNA technique trying to heal that specific disease. We are going to choose those which contain the mutated regions in their sequences for the chondrodysplasias. Once the oligonucleotides are selected they will be assayed in chondrocytes affected by different mutations or alternatively in transfected cells containing the mutated human FGFR3 for the different chondrodysplasias. The efficacy of the treatment will be checked by means of RT-PCR (to study the levels of nucleic acids), and also by immunocytochemistry and western blot to verify the presence or absence of the protein. Inhibition/reduction of signal transduction as a results of receptor silencing will be assessed in these cells, studying the major pathways related to FGFR3 (ERK, PI3K and STAT).
Complementarily, these cells will be studied in their fate in terms of maturation and also in their ability to produce/modify the composition of the extracellular matrix. Finally when the best siRNA are selected according to the results obtained in the previous experimentation, they will be injected into chondrodisplasic mice. The animals will be followed by measuring the most relevant morphometric parameters and the best siRNA will be recommended for future clinical trial studies. To complement our effort we will intend to develop a treatment by targeting FGFR3 with siRNAs.
Keywords / Chondrodysplasias, miRNA , siRNA, FGFR3 mutation.
Partners already involved / Universitat Di Verona (UDV)-Group of Dr Elio Liboi, GenPharmTox Biotech AG (GPT) (SME)- Dr. Johannes Doehmer, The Hebrew University (THU) -Group of Dr Efrat Monsonego Ornan, Agricultural Biotechnology Center (ABC)- Group of Dr. Burgyán József, University
Project budget (for the running projects) / nc / Budget reserved for SMEs / nc

Research topics

• HEALTH-2007-1.4-4: Development of emerging gene therapy tools and technologies for clinical application.

Profile of SME sought

Role / technology development, research
Country /region / Spain
Start of partnership / start-up phase
Expertise required / Experience in genome editing and repair (RNAi, site specific
recombination, etc.),
Pharmacokinetics