Development of lentiviral vectors for genetical engineering of stem cells and endothelial progenitor cells

Résumé

DEVELOPMENT OF LENTIVIRAL VECTORS FOR GENETICAL ENGINEERING OF STEM CELLS AND ENDOTHELIAL PROGENITOR CELLS #RESUMEN: Since the Endothelial Progenitor cells (EPCs) and Multipotential Adult Progenitor cells (MAPCs) are endowed with the capacity to home to the tumor vasculature, they might be used for cellular vehicles to deliver toxic material inside tumor and to exert antitumoural activity. For this goal, we developed an endothelial cells specific single inducible lentiviral vector, we generated an endothelial cells lineage-specific promoter (murine vascular Endothelial Cadherin promoter) to control trans-activator (rtTA2s-M2) expression, and synthesized a minimal monodirectional Poly (A)(60PA) site for the single inducible lentiviral vector. A reporter gene firefly Luciferase was driven by PTREalb inducible promoter. Two distinct expression cassettes were inserted to the late-generation, self-inactivating lentiviral vector. We compared several inducible lentiviral vector structures, we considered this structure (Two distinct expression cassettes )optimal because of its absence of leakiness in the uninduced state, and could achieve high luciferase expression (17.7~73-fold induction) in the resence of Doxycycline in several endotheüal cellules, when mVEcad specific promoter replaced by hPGK constitutive promoter, we could obtain slightly better induction factor (88~345-fold) but very low background expression. The lineage-specific single regulatable lentiviral vector may thus be useful for many applications in gene therapy research, could open the way of cell-gene therapy. The combination of cell with gene therapy may provide a new promising strategy for potential treatment of cancer.