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2017-05-16 - Colloque/Présentation - communication orale - Anglais - 1 page(s)

Andre Séverine , Daldal Fatima, Delcroix M., Verteneuil Sébastien, Muller Robert , Vander Elst Luce , Laurent Sophie , Burtea Carmen , "Blood-brain barrier crossing for improving brain access using a peptide able to bind a receptor-mediated transcytosis" in BMIC, Bruxelles, Belgique, 2017

  • Codes CREF : Histologie (DI3212), Neurophysiologie (DI3224), Sciences biomédicales (DI3200), Biologie moléculaire (DI3111), Imagerie médicale, radiologie, tomographie (DI3243), Biologie cellulaire (DI311D)
  • Unités de recherche UMONS : Chimie générale, organique et biomédicale (M108)
  • Instituts UMONS : Institut des Sciences et Technologies de la Santé (Santé), Institut des Biosciences (Biosciences)
  • Centres UMONS : Centre de Recherche en Microscopie et Imagerie Médicale (CMMI)

Abstract(s) :

(Anglais) Introduction The diagnosis and treatment of brain diseases is complicated by the presence of the blood-brain barrier (BBB), protecting against xenobiotics and limiting the access of most molecules, including potential therapeutic agents. This barrier, composed of endothelial cells joined by tight junctions, is estimated to restrict brain access of 98% of the blood-circulating molecules. Non-invasive crossing strategies are thus indispensable to accede to the central nervous system (CNS) without BBB disruption. The use of transcytosis-mediated receptors is a non-invasive natural pathway allowing the delivery of molecules across the BBB. In this context, LDL receptor (LDLR) turns out to be an attractive shuttling strategy for drug delivery because of its involvement in LDL transcytosis. Methods The phage display technology allows the identification of specific peptides against a target, here the LDLR. After selection steps, the potential as vector of the hit peptide, called LRP2, was evaluated. The endocytosis of LRP2 was assessed on human umbilical vein endothelial cells (HUVEC) and human brain endothelial cells (ACBRI376) by fluorescence. The colocalization of LRP2 with LDLR was validated by immunofluorescence on mouse brain slices and ACBRI376. The endocytosis pathway followed by LRP2 was investigated by fluorescent detection of caveolae and lysosomes in ACBRI376, astrocytoma 1321N1 and neuroblastoma N18(H) cells. Finally, LR-P2 was grafted to Ultrasmall Superparamagnetic Particles of Iron Oxide (USPIO-LRP2) and injected in mice to evaluate the BBB crossing by Magnetic Resonance Imaging (MRI) and histology. Results Peptide LRP2 showed a good endocytosis in both endothelial cell types and colocalized with LDLR on mouse brain slices, as well as in ACBRI376. LRP2 is endocytosed in ACBRI376 via a caveolae-mediated non-degradation pathway, whereas the lysosome degradation is bypassed. In 1321N1 and N18(H), LRP2 seems to be present in both compartments, although caveolae remains predominant. In vivo MRI experiments and contrast analysis performed on images revealed that USPIO-LRP2 is found in mouse brains compared to NSP (non-specific peptide) vectorized nanoparticles. These results were confirmed by Perls/DAB staining where USPIO-LRP2 was found within brain parenchyma, around the 3rd ventricle and brain capillaries. Conclusions Peptide LRP2 possesses a good potential to be used as vector through the BBB. It could operate as a delivery vehicle of various agents, including the therapeutic and diagnostic ones. In the near future, it will be used combined to a therapeutic peptide identified in the context of Alzheimer’s disease in order to improve its in vivo efficacy.