       Document 0594
 DOCN  M9550594
 TI    Sequence-specific inhibition of gene expression by a novel antisense
       oligodeoxynucleotide phosphorothioate directed against a nonregulatory
       region of the human immunodeficiency virus type 1 genome.
 DT    9505
 AU    Anazodo MI; Wainberg MA; Friesen AD; Wright JA; Manitoba Institute of
       Cell Biology, University of Manitoba,; Winnipeg, Canada.
 SO    J Virol. 1995 Mar;69(3):1794-801. Unique Identifier : AIDSLINE
       MED/95156611
 AB    Previous studies have demonstrated that oligodeoxynucleotide
       phosphorothioates complementary to human immunodeficiency virus type 1
       (HIV-1) RNA are more nuclease resistant and are effective inhibitors of
       HIV-1 replication than their unmodified counterpart. In this study,
       antisense oligodeoxynucleotide sequences were evaluated for therapeutic
       potential in the treatment of HIV infections. The use of HIV-infected
       lymphocytes to test the efficacy of a drug is very complex, and
       therefore it is difficult to draw conclusions about the mechanism. We
       used a COS-like Monkey kidney cell line (CMT3) stably transfected with
       plasmids pCMVgagpol-rre-r (containing gag and pol genes) and pCMVrev
       (containing the rev gene of HIV-1), derived from cDNA clone BH10, as a
       model. A biologically active provirus that transcribes and translates
       their nucleotide sequences into viral proteins p24, p39/41, p55, and
       p160 was generated. Sequence-specific and dose-dependent inhibition of
       HIV-1 viral protein synthesis and significant inhibition at the mRNA
       level were demonstrated by antisense construct GPI2A, directed against a
       nonregulatory region of the HIV-1 genome. Also, our studies demonstrated
       enhancement of the antisense effect through encapsulation in a cationic
       lipid preparation. The observed attenuation of HIV-1 mRNA levels
       suggests that, at least in part, the mechanism of action of GPI2A was at
       the transcript level. Further studies have also shown antiviral activity
       of this construct as determined by the reverse transcriptase assay using
       acutely and chronically infected cells of lymphoid origin (H9 cells).
       Toxicological studies involving cell growth characteristics,
       colony-forming ability, effects on cellular proteins, specific
       activities of labeled proteins, and DNA synthesis in cell culture showed
       no cytotoxic effects of GPI2A.
 DE    Animal  *Antiviral Agents  Base Sequence  Cations  Cell Line
       Cercopithecus aethiops  Gene Expression Regulation, Viral/*DRUG EFFECTS
       *Genes, gag  HIV Infections/*THERAPY  HIV-1/GROWTH &
       DEVELOPMENT/*GENETICS  In Vitro  Liposomes  Molecular Sequence Data
       Oligonucleotides, Antisense/*CHEMISTRY/PHARMACOLOGY
       Phosphatidylethanolamines/CHEMISTRY  RNA, Messenger/GENETICS  RNA,
       Viral/GENETICS  Support, Non-U.S. Gov't  Thionucleotides  Virus
       Replication/DRUG EFFECTS  JOURNAL ARTICLE

       SOURCE: National Library of Medicine.  NOTICE: This material may be
       protected by Copyright Law (Title 17, U.S.Code).

