Inhibition of Human Immunodeficiency Virus Replication by Cell Membrane-Crossing Oligomers
Although rapidly becoming a valuable tool for gene silencing, regulation or editing in vitro, the direct transfer of small interfering ribonucleic acids (siRNAs) into cells is still an unsolved problem for in vivo applications. For the first time, we show that specific modifications of antisense oligomers allow autonomous passage into cell lines and primary cells without further adjuvant or coupling to a cell-penetrating peptide. For this reason, we termed the specifically modified oligonucleotides “cell membrane-crossing oligomers” (CMCOs). CMCOs targeted to various conserved regions of human immunodeficiency virus (HIV)-1 were tested and compared with nontargeting CMCOs. Analyses of uninfected and infected cells incubated with labeled CMCOs revealed that the compounds were enriched in infected cells and some of the tested CMCOs exhibited a potent antiviral effect. Finally, the CMCOs did not exert any cytotoxicity and did not inhibit proliferation of the cells. In vitro, our CMCOs are promising candidates as biologically active anti-HIV reagents for future in vivo applications.
The authors were supported by the Austrian Science Found (FWF; project P22165 to D Wilflingseder), the Tyrolean Science Fund (TWF; project D-155140-016-011 to W Posch) and the Bridge Project (Austrian Research Promotion Agency [FFG]; project 815463 to H Stoiber). We would like to thank Christoph Gassner from the local blood bank in Innsbruck, Austria, and CIT Safety and Health Research Laboratories, France, for performing the CMCO stability tests. The reagent ARP033 (M8166 cells) was obtained from the Programme EVA Center for AIDS Reagents (NIBSC, UK) supported by the EC FP6/7 Europrise Network of Excellence, AIDS Vaccine Integrated Project (AVIP) and “Next Generation HIV-1 Immunogens inducing broadly reactive Neutralising antibodies” (NGIN) consortia and the Bill and Melinda Gates GHRC-CAVD Project and was donated by P Clapham. The CMCOs used in this study were already patented (Lindhorst et al. , WO 2008/009470; see http://www.wipo.int/patentscope/search/en/WO2008009470). T Lindhorst and H Bock are owners in ugichem GmbH.
- 18.Lindhorst T, Werner B, Bock H, inventors; ugichem GmbH, Lindhorst T, Werner B, Bock H, assignees. Chiral compounds substituted with phosphonate ester or phosphonic acid. World Intellectual Property Organization patent WO/2008/009470. 2008 Jan 24.Google Scholar
- 23.Schöllkopf U, Busse U, Lonsky R, Hinrichs R. (1986) Asymmetric syntheses via heterocyclic intermediates, XXXI: Asymmetric synthesis of various non-proteinogenic amino acid methyl esters (functionalized in the carbon chain) and amino acids by the bislactim ether method. Liebigs Annalen der Chemie. 12:2150–63.CrossRefGoogle Scholar
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