title: A polyethylenimine-linoleic acid conjugate for antisense oligonucleotide delivery.
abstract: A novel antisense oligonucleotide (ASO) carrier, polyethylenimine conjugated to linoleic acid (PEI-LA), was synthesized and evaluated for delivery of LOR-2501 to tumor cells. LOR-2501 is an ASO targeting ribonucleotide reductase R1 subunit (RRM1). In this study, PEI-LA was synthesized by reacting PEI (Mw ~ 800) with linoleoyl chloride. Gel retardation assay showed complete complexation between PEI-LA and LOR-2501 at N/P ratio above 8. No significant cytotoxicity was observed with these complexes at the tested dosage levels. Interestingly, at N/P ratio of >6, levels of cellular uptake of PEI-LA/LOR-2501 were double that of PEI/LOR-2501 complexes of the same N/P ratio. PEI-LA/LOR-2501 induced downregulation of 64% and 70% of RRM1 at mRNA and protein levels, respectively. The highest transfection activity was shown by PEI-LA/LOR-2501 complexes at N/P ratio of 10. Finally, using pathway specific inhibitors, clathrin-mediated endocytosis was shown to be the principle mechanism of cellular internalization of these complexes. In conclusion, PEI-LA is a promising agent for the delivery of ASOs and warrants further investigation.
authors.affiliation: Institute of Life Sciences, Jilin University, Changchun, Jilin 130023, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Xie
authors.forename: Lesheng
authors.initials: L
authors.lastname: Teng
authors.forename: Zhaogang
authors.initials: Z
authors.lastname: Yang
authors.forename: Chenguang
authors.initials: C
authors.lastname: Zhou
authors.forename: Yang
authors.initials: Y
authors.lastname: Liu
authors.forename: Bryant C
authors.initials: BC
authors.lastname: Yung
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Lee
mesh_terms: D015153:Blotting, Western; D016923:Cell Death / Q000187:drug effects; D002460:Cell Line; D015536:Down-Regulation / Q000187:drug effects / Q000235:genetics; D004587:Electrophoresis, Agar Gel; D005434:Flow Cytometry; D018014:Gene Transfer Techniques; D006801:Humans; D042541:Intracellular Space / Q000187:drug effects / Q000378:metabolism; D019787:Linoleic Acid / Q000737:chemistry*; D018613:Microscopy, Confocal; D016376:Oligonucleotides, Antisense / Q000378:metabolism* / Q000494:pharmacology; D011094:Polyethyleneimine / Q000737:chemistry*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D055672:Static Electricity
chemical_list: D016376:Oligonucleotides, Antisense; D012333:RNA, Messenger; D011094:Polyethyleneimine; D019787:Linoleic Acid