This Article Full Text Full Text (PDF) Submit a response Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Liu, K. Articles by Arakawa, K. Search for Related Content PubMed PubMed Citation Articles by Liu, K. Articles by Arakawa, K. Pubmed/NCBI databases Substance via MeSH Related Collections Gene therapy Lipid and lipoprotein metabolism Receptor pharmacology

(Arteriosclerosis, Thrombosis, and Vascular Biology. 1999;19:2207-2213.)
© 1999 American Heart Association, Inc.

Atherosclerosis and Lipoproteins

Efficient Nuclear Delivery of Antisense Oligodeoxynucleotides and Selective Inhibition of CETP Expression by Apo E Peptide in a Human CETP–Stably Transfected CHO Cell Line

Kan Liu; Jiafu Ou; Keijiro Saku; Shiro Jimi; David P. Via; James T. Sparrow; Bo Zhang; Henry J. Pownall; Louis C. Smith; Kikuo Arakawa

From the Departments of Internal Medicine (K.L., J.O., K.S., B.Z., K.A.) and Pathology (S.J.), Fukuoka University School of Medicine, Fukuoka, Japan, and the Department of Internal Medicine (D.P.V., J.T.S., H.J.P. L.C.S.), Baylor College of Medicine, Houston, Tex.

Correspondence to Keijiro Saku, MD, PhD, FACP, Department of Internal Medicine, Fukuoka University School of Medicine, 7-45-1 Nanakuma Jonan-ku, Fukuoka 814-80, Japan. E-mail hh035399{at}msat.fukuoka-u.ac.jp

Abstract—N,N-Dipalmitylglycyl–apolipoprotein E (129–169) peptide (dpGapoE) is an efficient gene delivery system for both plasmids and antisense oligodeoxynucleotides (ODNs). To develop a new and efficient approach to the regulation of cholesteryl ester transfer protein (CETP) expression, we used dpGapoE to transfect phosphorothioate antisense ODNs against nucleotides 329 to 349 of human CETP cDNA into a human CETP–stably transfected Chinese hamster ovary (CHO) cell line (hCETP-CHO). After transfection, translocation to the nuclei and concentration in nuclear structures were observed in >95% of the cells at 6 and 12 hours by fluorescence microscopy. No membrane disruption was observed after transfection of ODNs by dpGapoE. Although the translocation stability of phosphorothioate ODNs in the nuclei continued for >48 hours, it had weakened after 24 hours. Cellular CETP mRNA levels gradually declined, and the maximum reduction in the mRNA level (>50%) was observed at 36 hours, after which the mRNA level started to recover. CETP activity in the culture medium declined over 72 hours. The maximum reduction in CETP activity was observed at 36 hours (53.8% of control). Neither CETP mRNA nor CETP activities changed throughout the experiment after the transfection of sense phosphorothioate ODNs delivered by dpGapoE complex or naked antisense ODNs. We conclude that (1) the novel synthetic dpGapoE was a highly effective and nontoxic vehicle for the nuclear delivery of antisense ODNs into hCETP-CHO cells and (2) antisense ODNs selectively inhibited both CETP expression and activity in an hCETP-CHO cell line. This approach may enable gene regulation in vivo and could possibly be used as an antiatherosclerotic agent to alter high density lipoprotein metabolism.

Key Words: peptide vectors • receptor-mediated gene transfer • LDL receptor • gene therapy • in vitro studies

This article has been cited by other articles:

				R. W. Mahley, K. H. Weisgraber, and Y. Huang Inaugural Article: Apolipoprotein E4: A causative factor and therapeutic target in neuropathology, including Alzheimer's disease PNAS, April 11, 2006; 103(15): 5644 - 5651. [Abstract] [Full Text] [PDF]

				S. Chang, T. r. Ma, R. D. Miranda, M. E. Balestra, R. W. Mahley, and Y. Huang Lipid- and receptor-binding regions of apolipoprotein E4 fragments act in concert to cause mitochondrial dysfunction and neurotoxicity PNAS, December 20, 2005; 102(51): 18694 - 18699. [Abstract] [Full Text] [PDF]