Brain-targeting gene delivery and cellular internalization mechanisms for modified rabies virus glycoprotein RVG29 nanoparticles.

A 29 amino-acid peptide derived from the rabies virus glycoprotein (RVG29) was exploited as a ligand for efficient brain-targeting gene delivery. RVG29 was modified on polyamidoamine dendrimers (PAMAM) through bifunctional PEG, then complexed with DNA, yielding PAMAM-PEG-RVG29/DNA nanoparticles (NPs). The NPs were observed to be uptaken by brain capillary endothelial cells (BCECs) through a clathrin and caveolae mediated energy-depending endocytosis. The specific cellular uptake can be inhibited by free RVG29 and GABA but not by nicotinic acetylcholine receptor (nAchR) agonists/antagonists, indicating RVG29 probably relates to the GABA(B) receptor besides nAchR reported previously. PAMAM-PEG-RVG29/DNA NPs showed higher blood-brain barrier (BBB)-crossing efficiency than PAMAM/DNA NPs in an in vitro BBB model. In vivo imaging showed that the NPs were preferably accumulated in brain. The report gene expression of the PAMAM-PEG-RVG29/DNA NPs was observed in brain, and significantly higher than unmodified NPs. Thus, PAMAM-PEG-RVG29 provides a safe and noninvasive approach for the gene delivery across the BBB.

[1]  J. Baker,et al.  DNA complexing with polyamidoamine dendrimers: implications for transfection. , 1999, Bioconjugate chemistry.

[2]  D. Luo,et al.  Poly(ethylene glycol)-Conjugated PAMAM Dendrimer for Biocompatible, High-Efficiency DNA Delivery , 2002 .

[3]  R. Pagano,et al.  Glycosphingolipids Internalized via Caveolar-related Endocytosis Rapidly Merge with the Clathrin Pathway in Early Endosomes and Form Microdomains for Recycling* , 2003, The Journal of Biological Chemistry.

[4]  B. Davidson,et al.  Transvascular delivery of small interfering RNA to the central nervous system , 2007, Nature.

[5]  Xiao-bin Zhang,et al.  [Establishment of an in vitro model of brain-blood barrier]. , 2004, Beijing da xue xue bao. Yi xue ban = Journal of Peking University. Health sciences.

[6]  E. Giralt,et al.  The role of peptides in blood‐brain barrier nanotechnology , 2008, Journal of peptide science : an official publication of the European Peptide Society.

[7]  W. Pardridge,et al.  Comparison of in vitro and in vivo models of drug transcytosis through the blood-brain barrier. , 1990, The Journal of pharmacology and experimental therapeutics.

[8]  M. Lesniak.  Gene therapy for malignant glioma , 2006, Expert review of neurotherapeutics.

[9]  A. Florence,et al.  Release of DNA from dendriplexes encapsulated in PLGA nanoparticles. , 2005, International journal of pharmaceutics.

[10]  H. Mochizuki,et al.  Advances in gene therapy for movement disorders , 2008, Neurotherapeutics.

[11]  V. Labhasetwar,et al.  TAT-conjugated nanoparticles for the CNS delivery of anti-HIV drugs. , 2008, Biomaterials.

[12]  C. Gotti,et al.  Neuronal nicotinic receptors: from structure to pathology , 2004, Progress in Neurobiology.

[13]  M. Honer,et al.  γ-Aminobutyric Acid Type B Receptor Splice Variant Proteins GBR1a and GBR1b Are Both Associated with GBR2 in Situ and Display Differential Regional and Subcellular Distribution* , 1999, The Journal of Biological Chemistry.

[14]  T. Nagai,et al.  Transport of nerve growth factor encapsulated into liposomes across the blood-brain barrier: in vitro and in vivo studies. , 2005, Journal of controlled release : official journal of the Controlled Release Society.

[15]  A. Salem,et al.  Conjugation of polyamidoamine dendrimers on biodegradable microparticles for nonviral gene delivery. , 2007, Bioconjugate chemistry.

[16]  M. Gumbleton,et al.  Endocytosis at the blood–brain barrier: From basic understanding to drug delivery strategies , 2006, Journal of drug targeting.

[17]  S. Laffray,et al.  Dissociation and trafficking of rat GABAB receptor heterodimer upon chronic capsaicin stimulation , 2007, The European journal of neuroscience.

[18]  J. Glorioso,et al.  Herpes simplex virus RNAi and neprilysin gene transfer vectors reduce accumulation of Alzheimer's disease-related amyloid-β peptide in vivo , 2006, Gene Therapy.

[19]  T. Smart,et al.  Tracking Cell Surface GABAB Receptors Using an α-Bungarotoxin Tag* , 2008, Journal of Biological Chemistry.

[20]  Rongqin Huang,et al.  Efficient gene delivery targeted to the brain using a transferrin-conjugated polyethyleneglycol-modified polyamidoamine dendrimer , 2007 .

[21]  J. Glorioso,et al.  Herpes simplex virus RNAi and neprilysin gene transfer vectors reduce accumulation of Alzheimer's disease-related amyloid-beta peptide in vivo. , 2006, Gene therapy.

[22]  Wim E Hennink,et al.  Biodegradable polymers as non-viral carriers for plasmid DNA delivery. , 2008, Journal of controlled release : official journal of the Controlled Release Society.

[23]  M. Gassmann,et al.  Independent maturation of the GABAB receptor subunits GABAB1 and GABAB2 during postnatal development in rodent brain , 2004, The Journal of comparative neurology.

[24]  Shu Wang,et al.  An endosomolytic Tat peptide produced by incorporation of histidine and cysteine residues as a nonviral vector for DNA transfection. , 2008, Biomaterials.

[25]  T. Lentz,et al.  Is the acetylcholine receptor a rabies virus receptor? , 1982, Science.

[26]  J. Tolan,et al.  MDCK (Madin-Darby canine kidney) cells: A tool for membrane permeability screening. , 1999, Journal of pharmaceutical sciences.

[27]  V. Meskenaite,et al.  GABAB‐receptor splice variants GB1a and GB1b in rat brain: developmental regulation, cellular distribution and extrasynaptic localization , 1999, The European journal of neuroscience.

[28]  J. Huwyler,et al.  Uptake of Cationized Albumin Coupled Liposomes by Cultured Porcine Brain Microvessel Endothelial Cells and Intact Brain Capillaries , 2002, Journal of drug targeting.

[29]  Hideyoshi Harashima,et al.  Effect of transferrin receptor-targeted liposomal doxorubicin in P-glycoprotein-mediated drug resistant tumor cells. , 2007, International journal of pharmaceutics.

[30]  M. Ogris,et al.  Oligoethylenimine-grafted polypropylenimine dendrimers as degradable and biocompatible synthetic vectors for gene delivery. , 2008, Journal of controlled release : official journal of the Controlled Release Society.

[31]  K. Kent,et al.  Biodegradable arginine-based poly(ester-amide)s as non-viral gene delivery reagents. , 2008, Biomaterials.

[32]  R. Boado.  Blood–brain Barrier Transport of Non-viral Gene and RNAi Therapeutics , 2007, Pharmaceutical Research.

[33]  Chen Jiang,et al.  The use of lactoferrin as a ligand for targeting the polyamidoamine-based gene delivery system to the brain. , 2008, Biomaterials.

[34]  W. Pardridge,et al.  Gene therapy of the brain , 2004, Neurology.

[35]  Leaf Huang,et al.  Non-viral is superior to viral gene delivery. , 2007, Journal of controlled release : official journal of the Controlled Release Society.

[36]  M. Lafon.  Rabies virus receptors , 2008 .

[37]  W. Pardridge,et al.  Drug and Gene Delivery to the Brain The Vascular Route , 2002, Neuron.