Objetivo
For gene therapy, the use of non viral DNA offers the advantage of lack of immunogenicity, absence of DNA size limit, simpler GMP production, and improved safety/toxicity profiles. However, the efficient, precise and safe delivery of plasmids or of other forms of non viral DNA remains to be improved. Two different in vivo non viral DNA delivery techniques have been recently introduced, which are presently the most efficient: -Electrotransfer, using electric pulses (ET), -Laser beam gene transfer (LBGT). Our aim is to increase precision and selectivivity of non viral DNA transfer, by introducing molecular strategies to improve LT and LBGT. This optimization will be performed in skeletal muscle and skin, by assessing: -for each delivery mode (ET or LBGT), what is the best promoter (ubiquitous or tissue-specific), -the preferential form of non viral DNA: 1) conventional plasmid; 2) prokaryotic-backbone deleted "miniplasmid" devoid of antibiotic resistance gene; or 3) linear PCR-produced expression cassette, -the usefulness of an optimized secretion sequence for secreted transgenic proteins, -what is the best DNA formulation: "naked" versus associated to cationic or to non cationic lipid, -if pre- or post-iontophoresis is beneficial. This study will be performed in the context of 3 different gene therapy paradigms, and for 4 different medical applications: olong term intracellular expression of dystrophin gene in skeletal muscle, for the therapy of Duchenne muscular dystrophy, olong term blood secretion of circulating protein: erythropoietin (EPO) for chronic renal failure, and secreted monoclonal antibodies for antitumour passive immunization, oshort term transgene expression in skin for raising humoral and cellular immune response in antitumour vaccination. Our overall objective is to develop non viral DNA technology into a pre-clinical phase. Consequently, the safety issue (tissue damage, inflammation, integration, etc...) will be particularly...
Ámbito científico
- medical and health sciencesmedical biotechnologygenetic engineeringgene therapy
- medical and health sciencesbasic medicineimmunologyimmunisation
- natural sciencesbiological sciencesgeneticsDNA
- medical and health sciencesbasic medicineneurologymuscular dystrophiesduchenne muscular dystrophy
- medical and health sciencesbasic medicinepharmacology and pharmacydrug resistanceantibiotic resistance
Palabras clave
Convocatoria de propuestas
FP6-2003-LIFESCIHEALTH-I
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Régimen de financiación
IP - Integrated ProjectCoordinador
PARIS
Francia