PARTNERSHIP IN PRIORITARY DOMAINS
The period covered by the project: 2008 – 2011
Project Budget : 2.000.000
lei (Project financed by CNMP)
Coordinator: National Institute of Pathology “Victor Babes”
Project Director: Dr. Bogdan O. POPESCU, CS I
Biology Department, Molecular Medicine Laboratory, INCD “Victor Babes”
Tel.: 021-319.4528 e-mail: bogdan_ovidiu_popescu@yahoo.com
; bogdan.popescu@ivb.ro
Partners:
P1 – University of Bucharest, Biochemistry Dept., MURC-BM - Dr. Gisela Gaina, CS II
P2 – Emergency University Hospital from Bucharest - Prof. Dr. Ovidiu Alexandru Bajenaru
Key words: peripheral neuropathies, CMT, Schwann proteins, inflammtory neuropathies, demyelination
Peripheral neuropathies are peripheral nervous system diseases, with painful manifestations, stress, invalidity and even death. The diagnosis is many times difficult, and effective therapies are limited. Knowing the complexity of peripheral neuropathies pathology and the difficult diagnossis, or aim in the project is to investigate the peripheral nerve biopsy by new molecular methods beside the classical ones. We would like to bring new data concerning the mutations in the Schwann cell proteins gene, proteins implied in the maintaing of myelin sheath functioning. Thease data will improve the diagnosis, but would have a scientific value too.
We propose to study the interactions between these proteins in normal and pathological conditions, the litterature data being focusent on one or another problem, but not in a complex

view. An important aspect of the project will be the correlations between genotype and phenotype modifications, respectivelyvthe investigated protein expression in different peripheral neuropathies with demyelinating component. Because of the great importance of the myelin sheath component of the peripheral nerve, knowing that it is the first affected in the disease process and because the Schwann cells are the first that react in the axon regeneration and repair process, we thought to study the main Schwann proteins implied in inflammatory and hereditary demyelinating peripheral neuropathies: peripheral myelin protein 22 (PMP22) , myelin protein zero (MPZ or P0) , connexin 32 (Cx32), early growth response gene 2 (Egr 2), periaxin (PRX). Every of these proteins is essential for a normal myelination, and for the axon and Schwann cells interrelation. The protein expression is possibly modificated in axonal neuropathies and in the intermediate forms too, aspect that we propose to verify in this project.
Erythropoietin (Epo) is a protein that is not implied in a disease process, but we are interested in its study because it has a role in peripheral nerve reparatory action, in both normal and pathological state. The goal of our project is to find new molecular diagnosis methods for peripheral neuropaties and to study thoroughly some aspects of the interaction between the glial proteins in the peripheral nervous system for better understanding the pathological modifications occurrence and the nerve remyelination by Schwann cells. As a result of the study, valuable data which lead to a correct diagnostic in a very complex group of diseases will be obtained. The classical clinical and paraclinical methods are not sufficient to know the cause of demyelinating peripheral neuropathies. In this context, we appreciate that our project will clarify some aspects concerning at least the involvement of Schwann cell in the pathological processes of degenescence, but mostly regenerescence of the peripheral nerve in peripheral neuropathies.
Besides the practical appliance results, we anticipate scientific results concerning the interractions between the Schwann proteins for a normal cell function.
General objective: Schwann cell proteins in demyelinating peripheral neuropathies in scientific scope to investigate the connexion between them and in medical scope to improve the diagnosis in peripheral neuropathies patients. Oor study will help us to establish new directions in a future therapeutical research.

Specific Objectives:
1. To establish an experimental model of study the peripheral neuropathies with a demyelinating component by: determining the tasks for every partner in work meetings with the project partners leaders; establishing the subjects groups of study. It will be ellucidate ethical aspects regarding the subjects protection and the modality to take the nerve and muscle biopsy according with the current legislation.
2. Anatomopathological investigations of the nerve and muscle biopsies. The biopsied nerve is sural nerve, a sensitive one. Indirect data about the nerve will be given by the muscle fiber biopsy. It will be done also morphometrical investigations on nerve semithin sections. The teasing of nerve fibers will give us informations about myelin sheath. Electon microscpy studies on nerve ultrathin sections will be done also.
3. To introduce the fluorescent immunohistochemical technique (IHC), for establishing phenotype expression of the studied Schwann cell proteins from different patients. We will investigate the presence of erythropoietin in myelin sheath.
4. To introduce Western blotting technique (WB) for Schwann cell proteins expression identification (amount and quality).
5. To introduce MLPA technique for genetic analysis of main Schwann cell proteins genes.
6. To validate the experimental model. After all anatomopathological investigation methods and all molecular analysis methods for proteins were utilised, it will be done a complex study regarding the connexions between the modifications occuring in ageing muscle and a conclusion will be drawn regarding the experimental model proposed. The model will be implemented in diagnosis and in pathology investigations.
7. The development of knowledges in medical sciences, genetics and molecular biology.
8. The human resources formation.
9. To remove the differences between our country and european level in medical services domain, objectiv joining the 4th Program objectiv concerning the acceleration of the integration program in UE.
10. To form, improve, integrate and consolidate of a research network with an excellence level, according with international standards.
SCIENTIFIC MEETINGS

1.
Gisela Gaina, Aura Mihalcea, Emilia Manole, Elena Ionica - Molecular diagnoses of dystrophin gene mutation using MLPA , 1st International Conference 2008 of the
Cyprus Society of Human Genetics, Nicosia, Cyprus.
2.
Gisela Gaina , Emilia Manole, Elena Ionica
–
Molecular analyses of sarcolemal proteins in LGMD, al 6-lea Simpozion National de Patologie, “Patologie celulara si moleculara” si Sesiunea Anuala a Institutului National “Victor Babes”, 3-5 noiembrie
2009, Bucuresti
3.
E. Manole, G. Gaina, B. O. Popescu - General indications for the muscle biopsy, al
VIII-lea Congres al Societatii de Neurologie din Romania, 19-22 mai 2010, Bucuresti