title: Polymeric gene carrier for insulin secreting cells: poly(L-lysine)-g-sulfonylurea for receptor mediated transfection.
abstract: Ex vivo transfer of therapeutic genes to cells is one of the potential strategies to prolong the life span of cell transplants. However, relatively safe non-viral carriers have not been extensively investigated due to their lower transfection efficiency. In this study, poly(L-lysine)-g-sulfonylurea varying SU content (PLL-SU) was synthesized to promote gene delivery efficacy to an insulin secreting cell line, RINm5F, which is known to express sulfonylurea receptor (SUR). The polymer formed complexes with a model reporter gene of pCMV-Luc (DNA) and the size of resulting particles was around 100 nm. The transfection efficiency of a polymer synthesized with 5 mol% of SU in the reaction feed (PLL-SU5%) to RINm5F cell was at least 5 times higher than that of PLL. The cytotoxicity of PLL-SU5%/DNA complex was equivalent to that of PLL/DNA complex. PLL-SU5% showed less transfection efficiency than PLL to NIH3T3 and HepG2 cells which are SUR negative. In RINm5F cells, the addition of free SU decreased the transfection efficiency of PLL-SU5%/DNA complex, suggesting that the complex shares the same receptors for SU. The PLL-SU5%/DNA complex seems to be internalized via SUR-mediated endocytosis pathway as suggested by vacuolar ATPases inhibition by Bafilomycin A1. It is noted that RINm5F cells treated with PLL-SU5%/DNA complex secreted more insulin than control, untreated cells, suggesting the insulinotropic effect of SU in PLL-SU5%. In conclusion, PLL-SU may be useful for transfer of therapeutic genes into insulin secreting cells.
authors.affiliation: Department of Pharmaceutics and Pharmaceutical Chemistry, The University of Utah, 421 Wakara way, Suite 318, Salt Lake City, UT 84108, USA.
authors.forename: Han Chang
authors.initials: HC
authors.lastname: Kang
authors.forename: Sungwon
authors.initials: S
authors.lastname: Kim
authors.forename: Minhyung
authors.initials: M
authors.lastname: Lee
authors.forename: You Han
authors.initials: YH
authors.lastname: Bae
mesh_terms: D000818:Animals; D002470:Cell Survival / Q000187:drug effects; D004247:DNA / Q000737:chemistry / Q000235:genetics; D004337:Drug Carriers; D004563:Electrochemistry; D018390:Gene Targeting; D007515:Islets of Langerhans / Q000166:cytology / Q000378:metabolism*; D010316:Particle Size; D011107:Polylysine / Q000737:chemistry*; D011108:Polymers; D051381:Rats; D015203:Reproducibility of Results; D013453:Sulfonylurea Compounds / Q000737:chemistry*; D014162:Transfection
chemical_list: D004337:Drug Carriers; D011108:Polymers; D013453:Sulfonylurea Compounds; C505376:poly(L-lysine)-g-sulfonylurea; D011107:Polylysine; D004247:DNA