title: Rabies virus envelope glycoprotein targets lentiviral vectors to the axonal retrograde pathway in motor neurons.
abstract: Rabies pseudotyped lentiviral vectors have great potential in gene therapy, not least because of their ability to transduce neurons following their distal axonal application. However, very little is known about the molecular processes that underlie their retrograde transport and cell transduction. Using multiple labeling techniques and confocal microscopy, we demonstrated that pseudotyping with rabies virus envelope glycoprotein (RV-G) enabled the axonal retrograde transport of two distinct subtypes of lentiviral vector in motor neuron cultures. Analysis of this process revealed that these vectors trafficked through Rab5-positive endosomes and accumulated within a non-acidic Rab7 compartment. RV-G pseudotyped vectors were co-transported with both the tetanus neurotoxin-binding fragment and the membrane proteins thought to mediate rabies virus endocytosis (neural cell adhesion molecule, nicotinic acetylcholine receptor, and p75 neurotrophin receptor), thus demonstrating that pseudotyping with RV-G targets lentiviral vectors for transport along the same pathway exploited by several toxins and viruses. Using motor neurons cultured in compartmentalized chambers, we demonstrated that axonal retrograde transport of these vectors was rapid and efficient; however, it was not able to transduce the targeted neurons efficiently, suggesting that impairment in processes occurring after arrival of the viral vector in the soma is responsible for the low transduction efficiency seen in vivo, which suggests a novel area for improvement of gene therapy vectors.
authors.affiliation: From Gene Therapy, Centre for Neuroinflammation and Neurodegeneration, Division of Brain Sciences, Department of Medicine, Imperial College London, Du Cane Road, London W12 0NN, United Kingdom.
authors.forename: James N
authors.initials: JN
authors.lastname: Hislop
authors.affiliation: From Gene Therapy, Centre for Neuroinflammation and Neurodegeneration, Division of Brain Sciences, Department of Medicine, Imperial College London, Du Cane Road, London W12 0NN, United Kingdom.
authors.forename: Tarin A
authors.initials: TA
authors.lastname: Islam
authors.affiliation: From Gene Therapy, Centre for Neuroinflammation and Neurodegeneration, Division of Brain Sciences, Department of Medicine, Imperial College London, Du Cane Road, London W12 0NN, United Kingdom.
authors.forename: Ioanna
authors.initials: I
authors.lastname: Eleftheriadou
authors.affiliation: From Gene Therapy, Centre for Neuroinflammation and Neurodegeneration, Division of Brain Sciences, Department of Medicine, Imperial College London, Du Cane Road, London W12 0NN, United Kingdom.
authors.forename: David C J
authors.initials: DC
authors.lastname: Carpentier
authors.affiliation: From Gene Therapy, Centre for Neuroinflammation and Neurodegeneration, Division of Brain Sciences, Department of Medicine, Imperial College London, Du Cane Road, London W12 0NN, United Kingdom.
authors.forename: Antonio
authors.initials: A
authors.lastname: Trabalza
authors.affiliation: Molecular NeuroPathoBiology Laboratory, Cancer Research UK London Research Institute, Lincoln's Inn Fields Laboratories, 44 Lincoln's Inn Fields, London WC2A 3LY, United Kingdom, and.
authors.forename: Michael
authors.initials: M
authors.lastname: Parkinson
authors.affiliation: Molecular NeuroPathoBiology Laboratory, Cancer Research UK London Research Institute, Lincoln's Inn Fields Laboratories, 44 Lincoln's Inn Fields, London WC2A 3LY, United Kingdom, and Sobell Department of Motor Neuroscience and Movement Disorders, UCL Institute of Neurology, University College London, Queen Square, London WC1N 3BG, United Kingdom.
authors.forename: Giampietro
authors.initials: G
authors.lastname: Schiavo
authors.affiliation: From Gene Therapy, Centre for Neuroinflammation and Neurodegeneration, Division of Brain Sciences, Department of Medicine, Imperial College London, Du Cane Road, London W12 0NN, United Kingdom, n.mazarakis@imperial.ac.uk.
authors.forename: Nicholas D
authors.initials: ND
authors.lastname: Mazarakis
mesh_terms: D000818:Animals; D001370:Axonal Transport; D004705:Endocytosis; D005822:Genetic Vectors; D057809:HEK293 Cells; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D009046:Motor Neurons / Q000378:metabolism* / Q000821:virology; D011820:Rabies virus / Q000378:metabolism*; D051381:Rats; D014759:Viral Envelope Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D014759:Viral Envelope Proteins
keywords: Axon; Endosome; Glycoprotein; Lentivirus; Rab Proteins; Trafficking
references.citation: J Neurochem. 2012 May;121(4):575-86
references.pmid: 22243278
references.citation: PLoS Pathog. 2012 Dec;8(12):e1003087
references.pmid: 23300443
references.citation: Nat Med. 2005 Apr;11(4):429-33
references.pmid: 15768029
references.citation: Biochem J. 2012 May 1;443(3):603-18
references.pmid: 22507128
references.citation: Curr Top Microbiol Immunol. 2013;364:91-113
references.pmid: 23239350
references.citation: J Virol. 1993 Mar;67(3):1365-72
references.pmid: 8437221
references.citation: Curr Opin Cell Biol. 2008 Aug;20(4):445-53
references.pmid: 18495455
references.citation: J Virol. 1998 Sep;72(9):7181-90
references.pmid: 9696812
references.citation: Curr Opin Neurobiol. 2002 Jun;12(3):268-74
references.pmid: 12049932
references.citation: Dev Dyn. 1992 Jul;194(3):209-21
references.pmid: 1467557
references.citation: Future Virol. 2012 Jul 1;7(7):687-698
references.pmid: 23616788
references.citation: Nat Rev Microbiol. 2010 Sep;8(9):645-55
references.pmid: 20706281
references.citation: Virus Res. 2005 Jul;111(1):55-60
references.pmid: 15896402
references.citation: J Biol Eng. 2009 Jun 02;3:8
references.pmid: 19490632
references.citation: Neuron. 2006 Oct 19;52(2):293-305
references.pmid: 17046692
references.citation: Mol Ther. 2014 Jul;22(7):1285-1298
references.pmid: 24670531
references.citation: Infect Disord Drug Targets. 2012 Feb;12(1):38-58
references.pmid: 22034934
references.citation: J Cell Biol. 2002 Jan 21;156(2):233-9
references.pmid: 11807088
references.citation: PLoS Pathog. 2009 May;5(5):e1000442
references.pmid: 19461877
references.citation: Curr Gene Ther. 2005 Aug;5(4):387-98
references.pmid: 16101513
references.citation: Gene Ther. 2013 Jul;20(7):723-32
references.pmid: 23171919
references.citation: Eur J Neurosci. 2003 Feb;17(4):673-80
references.pmid: 12603257
references.citation: J Biol Chem. 2005 Dec 23;280(51):42336-44
references.pmid: 16236708
references.citation: Pflugers Arch. 2001 Sep;442(6):859-66
references.pmid: 11680618
references.citation: J Virol. 2008 Jan;82(1):237-45
references.pmid: 17928343
references.citation: Gene Ther. 2012 Jul;19(7):761-74
references.pmid: 21900965
references.citation: J Virol. 2000 Nov;74(21):10212-6
references.pmid: 11024151
references.citation: J Neurosci Methods. 2006 Oct 30;157(2):208-17
references.pmid: 16725205
references.citation: J Am Chem Soc. 2002 May 29;124(21):6063-76
references.pmid: 12022841
references.citation: Exp Cell Res. 2002 Jan 1;272(1):8-14
references.pmid: 11740860
references.citation: Exp Mol Pathol. 2011 Dec;91(3):723-32
references.pmid: 21819978
references.citation: Hum Gene Ther. 2010 May;21(5):589-602
references.pmid: 20039782
references.citation: Traffic. 2007 Dec;8(12):1736-1749
references.pmid: 17897318
references.citation: J Neurovirol. 2005 Feb;11(1):82-7
references.pmid: 15804965
references.citation: J Cell Biol. 2008 Feb 11;180(3):549-61
references.pmid: 18250195
references.citation: Gene. 1997 Feb 20;186(1):29-35
references.pmid: 9047341
references.citation: J Virol. 2014 Mar;88(5):2877-90
references.pmid: 24371049
references.citation: J Clin Invest. 2004 Dec;114(12):1726-31
references.pmid: 15599397
references.citation: Hum Gene Ther. 2006 Jan;17(1):1-9
references.pmid: 16409120
references.citation: EMBO J. 1998 Dec 15;17(24):7250-9
references.pmid: 9857182
references.citation: PLoS One. 2013 Jun 28;8(6):e67400
references.pmid: 23840690
references.citation: Gene. 1989 Apr 15;77(1):61-8
references.pmid: 2744488
references.citation: Cell. 2005 Sep 9;122(5):735-49
references.pmid: 16143105
references.citation: Proc Natl Acad Sci U S A. 2012 Oct 23;109(43):17627-32
references.pmid: 23047692
references.citation: J Virol. 2004 Oct;78(19):10803-13
references.pmid: 15367647
references.citation: J Cell Biol. 2006 Jul 31;174(3):459-71
references.pmid: 16880274
references.citation: Nature. 2004 May 27;429(6990):413-7
references.pmid: 15164063
references.citation: J Virol. 2005 Apr;79(7):4055-65
references.pmid: 15767407
references.citation: EMBO J. 2011 Aug 31;30(17):3527-39
references.pmid: 21878994
references.citation: Biol Chem. 2009 Aug;390(8):753-9
references.pmid: 19361275
references.citation: J Cell Biol. 2007 Feb 12;176(4):459-71
references.pmid: 17283181
references.citation: Hum Mol Genet. 2001 Sep 15;10(19):2109-21
references.pmid: 11590128
references.citation: Yale J Biol Med. 1983 Jul-Aug;56(4):315-22
references.pmid: 6367238
references.citation: J Cell Biol. 2002 Nov 11;159(3):441-52
references.pmid: 12417576
references.citation: J Gene Med. 2004 Feb;6 Suppl 1:S83-94
references.pmid: 14978753
references.citation: Nat Rev Microbiol. 2010 Jan;8(1):51-61
references.pmid: 19946287
references.citation: Nat Rev Mol Cell Biol. 2001 Feb;2(2):107-17
references.pmid: 11252952
references.citation: Mol Biol Cell. 2007 Aug;18(8):3204-13
references.pmid: 17567954
references.citation: Science. 1998 Jul 10;281(5374):269-72
references.pmid: 9657724
references.citation: Mol Membr Biol. 1999 Jan-Mar;16(1):21-31
references.pmid: 10332734
references.citation: Virology. 2007 Dec 5;369(1):78-91
references.pmid: 17707875