title: Delivery of therapeutic AGT shRNA by PEG-Bu for hypertension therapy.
abstract: Gene silencing by RNA interference (RNAi) is a promising approach for gene therapy. However, up to today, it is still a major challenge to find safe and efficient non-viral vectors. Previously, we reported PEI-Bu, a small molecular weight PEI derivative, as an efficient non-viral carrier. However, like many PEI-based polymers, PEI-Bu was too toxic. In order to reduce cytotoxicity while maintain or even enhance transfecion efficiency, we modified PEI-Bu with poly(ethylene glycol) (PEG) to obtain PEG-Bu, and used it to delivery a theraputic short hairpin RNA (shRNA) targeting angiotensinogen (AGT) to normal rat liver cells (BRL-3A), which was a key target for the treatment of hypertension. The structure of PEG-Bu was confirmed by proton nuclear magnetic resonance ((1)H-NMR). Gel permeation chromatography (GPC) showed that the weight average molecular weight (Mw) of PEG-Bu was 5880 Da, with a polydispersity of 1.58. PEG-Bu could condense gene cargo into spherical and uniform nanoparticles with particle size (65-88 nm) and zeta potential (7.3-9.6 mV). Interestingly and importantly, PEG-Bu displayed lower cytotoxicity and enhanced tranfection efficiency than PEI-Bu after PEGylation in both normal cells BRL-3A and tumor cells HeLa. Moreover, PEG-Bu could efficiently delivery AGT shRNA to knockdown the AGT expression. To sum up, PEG-Bu would be a promising non-viral vector for delivering AGT shRNA to BRL-3A cells for hypertension therapy.
authors.affiliation: Department of Geriatrics, Xinhua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai, China.
authors.forename: Yu-Qiang
authors.initials: YQ
authors.lastname: Wang
authors.forename: Fei
authors.initials: F
authors.lastname: Wang
authors.forename: Xiao-Qin
authors.initials: XQ
authors.lastname: Deng
authors.forename: Jing
authors.initials: J
authors.lastname: Sheng
authors.forename: Shu-Yan
authors.initials: SY
authors.lastname: Chen
authors.forename: Jing
authors.initials: J
authors.lastname: Su
mesh_terms: D000808:Angiotensinogen / Q000235:genetics*; D000818:Animals; D002460:Cell Line; D020868:Gene Silencing; D018014:Gene Transfer Techniques; D006801:Humans; D006973:Hypertension / Q000235:genetics* / Q000628:therapy; D007097:Imines / Q000737:chemistry / Q000633:toxicity; D008970:Molecular Weight; D010957:Plasmids / Q000737:chemistry / Q000235:genetics; D011095:Polyethylenes / Q000737:chemistry / Q000633:toxicity; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000008:administration & dosage / Q000235:genetics*; D051381:Rats; D014162:Transfection
chemical_list: D007097:Imines; D011095:Polyethylenes; D034741:RNA, Small Interfering; C505405:poly(ethylene imine); D000808:Angiotensinogen
references.citation: Eur J Cell Biol. 2004 Apr;83(3):97-111
references.pmid: 15202568
references.citation: Drug Des Devel Ther. 2013;7:211-21
references.pmid: 23576866
references.citation: Hypertension. 1997 Dec;30(6):1325-30
references.pmid: 9403548
references.citation: Proc Natl Acad Sci U S A. 2007 Oct 9;104(41):16050-5
references.pmid: 17913877
references.citation: J Drug Target. 2000;8(1):51-66
references.pmid: 10761645
references.citation: J Biol Chem. 1994 Dec 16;269(50):31334-7
references.pmid: 7989296
references.citation: Gene Ther. 1997 Sep;4(9):950-60
references.pmid: 9349432
references.citation: Biotechniques. 2001 Jan;30(1):74-7, 80-1
references.pmid: 11196323
references.citation: EMBO J. 1992 Mar;11(3):821-7
references.pmid: 1547785
references.citation: J Control Release. 2007 Dec 4;124(1-2):69-80
references.pmid: 17897749
references.citation: Biotechnol Appl Biochem. 2003 Jun;37(Pt 3):267-71
references.pmid: 12597775
references.citation: Curr Opin Mol Ther. 2009 Apr;11(2):165-78
references.pmid: 19330722
references.citation: J Control Release. 2005 Nov 28;108(2-3):557-67
references.pmid: 16253376
references.citation: Biochem Biophys Res Commun. 2008 Mar 21;367(4):874-80
references.pmid: 18201560
references.citation: J Biomed Mater Res A. 2008 Mar 1;84(3):795-804
references.pmid: 17635020
references.citation: Int J Nanomedicine. 2012;7:693-704
references.pmid: 22359448
references.citation: Biomaterials. 2008 Jun;29(16):2535-47
references.pmid: 18316120
references.citation: Bioconjug Chem. 2002 Jul-Aug;13(4):845-54
references.pmid: 12121141
references.citation: Biomaterials. 2012 Sep;33(27):6520-32
references.pmid: 22695070
references.citation: J Control Release. 2010 Aug 3;145(3):272-80
references.pmid: 20403398
references.citation: Biomacromolecules. 2005 Nov-Dec;6(6):3440-8
references.pmid: 16283777
references.citation: Bioconjug Chem. 2003 Mar-Apr;14(2):473-9
references.pmid: 12643759
references.citation: Chem Rev. 2009 Feb;109(2):259-302
references.pmid: 19053809
references.citation: Curr Drug Targets. 2010 Mar;11(3):345-60
references.pmid: 20210759
references.citation: Pharm Res. 2006 Aug;23(8):1868-76
references.pmid: 16845585
references.citation: Adv Drug Deliv Rev. 1998 Mar 2;30(1-3):85-95
references.pmid: 10837604
references.citation: J Renin Angiotensin Aldosterone Syst. 2012 Jun;13(2):282-8
references.pmid: 22156763