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Mechanisms of in vivo Electro-Permeabilization

Electropermeabilization of tissues in living mice

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Tissues in living mice such as tumors, muscles and skin are more complex than cells or spheroids. We have already shown in vivo an efficient electrotransfer of different types of molecules (DNA, shRNA, siRNA, morpholinos, cis-platinum, bleomycin…)

 

Following of tumor growth and treatment evaluation Electro-chemotherapy: S.C. tumors. Golzio M et al., Biochem Biophys Res Commun. 2006 

 

In this context, we have studied gene electrotransfer in mouse muscle by EP (effect of the electric parameters, expression and bio-distribution of the transgene upon the polarity of the field)

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Following of gene transfert (GFP) in muscle. Faurie C., Golzio M., et al. DNA and Cell Biology 2003

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Gene expression regulation by RNA interference (Subcutaneous B16F10-GFP tumor / electrotransfer). Golzio M. et al., Gene Ther 2007 

Challenges

Tissues in living mice such as tumors, muscles and skin are of interest for therapeutic purposes in cutaneous cancers (melanoma and sarcoids) but also for DNA vaccination. The bio-distribution, the stability and the route of administration of these molecules are crucial aspects that must be determined for clinical applications as well as inflammatory response induced by electric field and the implication of the immune system. In this context, the development of DNA vaccine strategies will be evaluated after intra muscular or intradermal injection.

Investigators : Postdoc: L. Pasquet / Project leader: M. Golzio

Collaborations : B. Couderc (ICR, Toulouse), M. Cemazar (Institut of Oncology, Ljubljana)

Blood vessels permeability induced by EP

In vivo EP has also blood flow modifying effect. A decreased blood flow (vascular lock) was observed in muscles and tumors. Our aim is to evaluate the effects of EP on subcutaneous blood vessels morphology (diameter) and dynamics (vasomotricity, permeability and recovery). These features are measured using fluorescently labeled dextran via dorsal skin fold window chamber, intravital fluorescence microscopy imaging and specific image analysis.

 

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Vascular lock induced by electroporation compared with normal vessels filling in control

We also observed that the permeability of the blood vessels is affected by the electric field.

 

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Blood vessels permeability induced by electroporation compared with control. (Bellard et al., J Control Release 2012) 

Challenges

Further experiments will be done to characterize the life-time of these processes in normal but also tumor vessels. In order to study these processes, we developed and used fluorescence macro-scopes combined or not with intravital microscopy (conventional or multiphoton microscopy). Thus, we can follow cellular or molecular events on the living animals (from minutes, to months).

Investigators : Postdoc: B. Markelc / Engineer: E. Bellard / Project leader: M. Golzio

Collaborations : M. Cemazar (Institut of Oncology, Ljubljana)