Plasmid size up to 20 kbp does not limit effective in vivo lung gene transfer using compacted DNA nanoparticles

被引:110
作者
Fink, T. L.
Klepcyk, P. J.
Oette, S. M.
Gedeon, C. R.
Hyatt, S. L.
Kowalczyk, T. H.
Moen, R. C.
Cooper, M. J.
机构
[1] Copernicus Therapeut Inc, Cleveland, OH 44106 USA
[2] Case Western Reserve Univ, Cleveland, OH 44106 USA
关键词
non-viral; nanoparticle; polyplex; lung; cystic fibrosis;
D O I
10.1038/sj.gt.3302761
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Nanoparticles consisting of single molecules of DNA condensed with polyethylene glycol-substituted lysine 30-mers efficiently transfect lung epithelium following intrapulmonary administration. Nanoparticles formulated with lysine polymers having different counterions at the time of DNA mixing have distinct geometric shapes: trifluoroacetate or acetate counterions produce ellipsoids or rods, respectively. Based on intracytoplasmic microinjection studies, nanoparticle ellipsoids having a minimum diameter less than the 25 nm nuclear membrane pore efficiently transfect nondividing cells. This 25 nm size restriction corresponds to a 5.8 kbp plasmid when compacted into spheroids, whereas the 8-11 nm diameter of rod-like particles is smaller than the nuclear pore diameter. In mice, up to 50% of lung cells are transfected after dosing with a rod-like compacted 6.9 kbp lacZ expression plasmid, and correction of the CFTR chloride channel was observed in humans following intranasal administration of a rod-like compacted 8.3 kbp plasmid. To further investigate the potential size and shape limitations of DNA nanoparticles for in vivo lung delivery, reporter gene activity of ellipsoidal and rod-like compacted luciferase plasmids ranging in size between 5.3 and 20.2 kbp was investigated. Equivalent molar reporter gene activities were observed for each formulation, indicating that microinjection size limitations do not apply to the in vivo gene transfer setting.
引用
收藏
页码:1048 / 1051
页数:4
相关论文
共 8 条
[1]   INHIBITION OF NUCLEAR ACCUMULATION OF KARYOPHILIC PROTEINS IN LIVING CELLS BY MICROINJECTION OF THE LECTIN WHEAT-GERM AGGLUTININ [J].
DABAUVALLE, MC ;
SCHULZ, B ;
SCHEER, U ;
PETERS, R .
EXPERIMENTAL CELL RESEARCH, 1988, 174 (01) :291-296
[2]   Compacted DNA nanoparticles administered to the nasal mucosa of cystic fibrosis subjects are safe and demonstrate partial to complete cystic fibrosis transmembrane regulator reconstitution [J].
Konstan, MW ;
Davis, PB ;
Wagener, JS ;
Hilliard, KA ;
Stern, RC ;
Milgram, LJH ;
Kowalczyk, TH ;
Hyatt, SL ;
Fink, TL ;
Gedeon, CR ;
Oette, SM ;
Payne, JM ;
Muhammad, O ;
Ziady, AG ;
Moen, RC ;
Cooper, MJ .
HUMAN GENE THERAPY, 2004, 15 (12) :1255-1269
[3]  
Kowalczyk T, 2001, MOL THER, V3, pS359
[4]  
Kube D, 2003, MOL THER, V7, pS371
[5]   Nanoparticles of compacted DNA transfect postmitotic cells [J].
Liu, G ;
Li, DS ;
Pasumarthy, MK ;
Kowalczyk, TH ;
Gedeon, CR ;
Hyatt, SL ;
Payne, JM ;
Miller, TJ ;
Brunovskis, P ;
Fink, TL ;
Muhammad, O ;
Moen, RC ;
Hanson, RW ;
Cooper, MJ .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2003, 278 (35) :32578-32586
[6]   Size-dependent DNA mobility in cytoplasm and nucleus [J].
Lukacs, GL ;
Haggie, P ;
Seksek, O ;
Lechardeur, D ;
Freedman, N ;
Verkman, AS .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2000, 275 (03) :1625-1629
[7]   NUCLEAR-PROTEIN IMPORT - SPECIFICITY FOR TRANSPORT ACROSS THE NUCLEAR-PORE [J].
WOLFF, B ;
WILLINGHAM, MC ;
HANOVER, JA .
EXPERIMENTAL CELL RESEARCH, 1988, 178 (02) :318-334
[8]   Transfection of airway epithelium by stable PEGylated poly-L-lysine DNA nanoparticles in vivo [J].
Ziady, AG ;
Gedeon, CR ;
Miller, T ;
Quan, W ;
Payne, JM ;
Hyatt, SL ;
Fink, TL ;
Muhammad, O ;
Oette, S ;
Kowalczyk, T ;
Pasumarthy, MK ;
Moen, RC ;
Cooper, MJ ;
Davis, PB .
MOLECULAR THERAPY, 2003, 8 (06) :936-947