title: Construction of a pIX-modified Adenovirus Vector Able to Effectively Bind to Nanoantibodies for Targeting.
abstract: Current targeting strategies for genetic vectors imply the creation of a specific vector for every targeted receptor, which is time-consuming and expensive. Therefore, the development of a universal vector system whose surface can specifically bind molecules to provide efficient targeting is of particular interest. In this study, we propose a new approach in creating targeted vectors based on the genome of human adenovirus serotype 5 carrying the modified gene of the capsid protein pIX (Ad5-EGFP-pIX-ER): recombinant pseudoadenoviral nanoparticles (RPANs). The surfaces of such RPANs are able to bind properly modified chimeric nanoantibodies that specifically recognize a particular target antigen (carcinoembryonic antigen (CEA)) with high affinity. The efficient binding of nanoantibodies (aCEA-RE) to the RPAN capsid surfaces has been demonstrated by ELISA. The ability of the constructed vector to deliver target genes has been confirmed by experiments with the tumor cell lines A549 and Lim1215 expressing CEA. It has been shown that Ad5-EGFP-pIX-ER carrying aCEA-RE on its surface penetrates into the tumor cell lines A549 and Lim1215 via the CAR-independent pathway three times more efficiently than unmodified RPAN and Ad5-EGFP-pIX-ER without nanoantibodies on the capsid surface. Thus, RPAN Ad5-EGFP-pIX-ER is a universal platform that may be useful for targeted gene delivery in specific cells due to "nanoantibody-modified RPAN" binding.
authors.affiliation: N.F. Gamaleya Research Institute of Epidemiology and Microbiology, Ministry of Health of the Russian Federation; Gamaleya Str., 18, Moscow, Russia, 123098.
authors.forename: M N
authors.initials: MN
authors.lastname: Garas
authors.affiliation: Institute of Gene Biology, Russian Academy of Sciences; Vavilova Str., 34/5, Moscow, Russia, 119334.
authors.forename: S V
authors.initials: SV
authors.lastname: Tillib
authors.affiliation: N.F. Gamaleya Research Institute of Epidemiology and Microbiology, Ministry of Health of the Russian Federation; Gamaleya Str., 18, Moscow, Russia, 123098.
authors.forename: O V
authors.initials: OV
authors.lastname: Zubkova
authors.affiliation: N.F. Gamaleya Research Institute of Epidemiology and Microbiology, Ministry of Health of the Russian Federation; Gamaleya Str., 18, Moscow, Russia, 123098 ; K.I. Skryabin Moscow State Academy of Veterinary Medicine and Biotehnology, Akademik Skraybin Str., 23, Moscow, Russia, 109472.
authors.forename: V N
authors.initials: VN
authors.lastname: Rogozhin
authors.affiliation: Institute of Gene Biology, Russian Academy of Sciences; Vavilova Str., 34/5, Moscow, Russia, 119334.
authors.forename: T I
authors.initials: TI
authors.lastname: Ivanova
authors.affiliation: Institute of Gene Biology, Russian Academy of Sciences; Vavilova Str., 34/5, Moscow, Russia, 119334.
authors.forename: L A
authors.initials: LA
authors.lastname: Vasilev
authors.affiliation: N.F. Gamaleya Research Institute of Epidemiology and Microbiology, Ministry of Health of the Russian Federation; Gamaleya Str., 18, Moscow, Russia, 123098.
authors.forename: D Yu
authors.initials: DY
authors.lastname: Logunov
authors.affiliation: N.F. Gamaleya Research Institute of Epidemiology and Microbiology, Ministry of Health of the Russian Federation; Gamaleya Str., 18, Moscow, Russia, 123098.
authors.forename: M M
authors.initials: MM
authors.lastname: Shmarov
authors.affiliation: N.F. Gamaleya Research Institute of Epidemiology and Microbiology, Ministry of Health of the Russian Federation; Gamaleya Str., 18, Moscow, Russia, 123098.
authors.forename: I L
authors.initials: IL
authors.lastname: Tutykhina
authors.affiliation: N.F. Gamaleya Research Institute of Epidemiology and Microbiology, Ministry of Health of the Russian Federation; Gamaleya Str., 18, Moscow, Russia, 123098.
authors.forename: I B
authors.initials: IB
authors.lastname: Esmagambetov
authors.affiliation: N.F. Gamaleya Research Institute of Epidemiology and Microbiology, Ministry of Health of the Russian Federation; Gamaleya Str., 18, Moscow, Russia, 123098.
authors.forename: I Yu
authors.initials: IY
authors.lastname: Gribova
authors.affiliation: N.F. Gamaleya Research Institute of Epidemiology and Microbiology, Ministry of Health of the Russian Federation; Gamaleya Str., 18, Moscow, Russia, 123098.
authors.forename: A S
authors.initials: AS
authors.lastname: Bandelyuk
authors.affiliation: N.F. Gamaleya Research Institute of Epidemiology and Microbiology, Ministry of Health of the Russian Federation; Gamaleya Str., 18, Moscow, Russia, 123098.
authors.forename: B S
authors.initials: BS
authors.lastname: Naroditsky
authors.affiliation: N.F. Gamaleya Research Institute of Epidemiology and Microbiology, Ministry of Health of the Russian Federation; Gamaleya Str., 18, Moscow, Russia, 123098.
authors.forename: A L
authors.initials: AL
authors.lastname: Gintsburg
keywords: CEA; adenoviral vector; leucine zipper; nanobody; pIX
references.citation: J Gen Virol. 1979 Sep;44(3):783-800
references.pmid: 93618
references.citation: Mol Gen Mikrobiol Virusol. 2002;(2):30-5
references.pmid: 12180023
references.citation: J Virol. 2004 Apr;78(7):3470-9
references.pmid: 15016870
references.citation: J Virol. 2003 Dec;77(24):12931-40
references.pmid: 14645549
references.citation: Nat Methods. 2006 Nov;3(11):887-9
references.pmid: 17060912
references.citation: EMBO J. 1987 Jun;6(6):1733-9
references.pmid: 3038533
references.citation: J Virol. 1997 Aug;71(8):6204-7
references.pmid: 9223518
references.citation: PLoS One. 2011 Feb 09;6(2):e16792
references.pmid: 21347423
references.citation: Anticancer Res. 2009 Aug;29(8):2997-3001
references.pmid: 19661307
references.citation: Gene Ther. 2005 Jan;12(2):187-93
references.pmid: 15526007
references.citation: J Bacteriol. 1999 Aug;181(15):4611-6
references.pmid: 10419961
references.citation: Nature. 1993 Jun 3;363(6428):446-8
references.pmid: 8502296
references.citation: Curr Opin Biotechnol. 1999 Oct;10(5):440-7
references.pmid: 10508634
references.citation: J Biol Chem. 2004 Dec 10;279(50):51965-72
references.pmid: 15459193
references.citation: Antiviral Res. 2013 Mar;97(3):318-28
references.pmid: 23274786
references.citation: Acta Naturae. 2010 Jul;2(3):85-93
references.pmid: 22649655
references.citation: J Gen Virol. 2005 Jun;86(Pt 6):1581-1588
references.pmid: 15914835
references.citation: J Virol. 2002 Jul;76(14):6893-9
references.pmid: 12072490
references.citation: Anticancer Res. 2012 Aug;32(8):3137-40
references.pmid: 22843884
references.citation: J Virol. 2005 Mar;79(6):3382-90
references.pmid: 15731232
references.citation: J Gene Med. 2001 Nov-Dec;3(6):550-9
references.pmid: 11778901
references.citation: Acta Naturae. 2011 Jul;3(3):64-70
references.pmid: 22649695
references.citation: Mol Ther. 2003 Nov;8(5):813-21
references.pmid: 14599815
references.citation: Protein Sci. 2001 Mar;10(3):649-55
references.pmid: 11344333
references.citation: PLoS One. 2009 Dec 21;4(12):e8355
references.pmid: 20027223
references.citation: Antiviral Res. 2013 Mar;97(3):245-54
references.pmid: 23274623
references.citation: Mol Ther. 2004 Jun;9(6):942-54
references.pmid: 15194061
references.citation: Mol Cell Biol. 1991 Oct;11(10):4863-75
references.pmid: 1922023
references.citation: Adv Immunol. 2001;79:261-96
references.pmid: 11680009
references.citation: J Virol. 2005 Oct;79(19):12125-31
references.pmid: 16160140
references.citation: Endocr Relat Cancer. 2011 Apr 02;18(3):311-21
references.pmid: 21389059
references.citation: J Invest Dermatol. 1998 Jun;110(6):867-71
references.pmid: 9620291
references.citation: J Virol. 1999 Jun;73(6):5156-61
references.pmid: 10233980
references.citation: Virology. 2008 Aug 1;377(2):391-400
references.pmid: 18570971