Naslov
Receptor-mediated targeting of toxin results in selection of genetically protected hepatocytes without, bystander toxicity

Autori
Volarević, Martina ; Wu, Catherine H. ; Smolić, Robert ; Andorfer, John H. ; Wu, George Y.

Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, znanstveni

Izvornik
FASEB JOURNAL / Gerald Weissmann, MD - Bethesda (MD) : Federation of American Societies for Experimental Biology (FASEB), 2007, A1137-A1137

Skup
Experimental Biology 2007

Mjesto i datum
Sjedinjene Američke Države, 28.04.2007. - 02.05.2007

Vrsta sudjelovanja
Poster

Vrsta recenzije
Međunarodna recenzija

Ključne riječi
asialoglycoprotein receptors (AsGR) ; G418 ; asialoorosomucoid (AsOR) ; GFP-Huh cells

Sažetak
ABSTRACT AIM To target G418 specifically to liver cells via asialoglycoprotein receptors (AsGR) for selection of stably transfected neomycin resistant hepatocytes (NR+) without collateral damage. METHODS: A novel G418 conjugate was synthesized by peptide coupling to a galactose-terminal asialoorosomucoid (AsOR). Uptake studies were done by labeled-ligand assays, and toxicity by MTT assays on Huh7 (AsGR+, NR– ), GFP-Huh7 (AsGR+, NR+), and 3T3 (AsGR– , NR– ) cell lines. RESULTS: Uptake of conjugate (7– 8 moles) of G418 per AsOR was the same as for AsOR alone. Conjugate inhibited growth of Huh7 cells by 84.3%. This inhibition was blocked by excess free AsOR. Incubation of 3T3 cells with free G418, but not conjugated G418 resulted in toxicity. GFP-Huh cells were not susceptible either to free or conjugated G418. Co-cultures of GFP-Huh7 and 3T3 in G418 alone resulted in only inhibition of 3T3 cells. In contrast, co-cultures with AsOR-G418 resulted in no significant toxicity to either cell type. CONCLUSIONS: G418-protein conjugates retained inhibitory activity targeted against AsGR (+), NR (– ) cells. G418 modification to form a conjugate did not block its activity. Lack of effect of G418 conjugate on ASGR (– ) cells suggests that the effects were specific, and not due to release of free G418 in the medium. The presence of NR gene in AsGR (+) cells protected those cells, and the effect without collateral damage to co-cultured AsGR (– ) cells.

Izvorni jezik
Engleski

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