Eliciting the low-activity aldehyde dehydrogenase Asian phenotype by an antisense mechanism results in an aversion to ethanol.
A mutation in the gene encoding for the liver mitochondrial aldehyde dehydrogenase (ALDH2-2), present in some Asian populations, lowers or abolishes the activity of this enzyme and results in elevations in blood acetaldehyde upon ethanol consumption, a phenotype that greatly protects against alcohol abuse and alcoholism. We have determined whether the administration of antisense phosphorothioate oligonucleotides (ASOs) can mimic the low-activity ALDH2-2 Asian phenotype. Rat hepatoma cells incubated for 24 h with an antisense oligonucleotide (ASO-9) showed reductions in ALDH2 mRNA levels of 85% and ALDH2 (half-life of 22 h) activity of 55% equivalent to a >90% inhibition in ALDH2 synthesis. Glutamate dehydrogenase mRNA and activity remained unchanged. Base mismatches in the oligonucleotide rendered ASO-9 virtually inactive, confirming an antisense effect. Administration of ASO-9 (20 mg/kg/day for 4 d) to rats resulted in a 50% reduction in liver ALDH2 mRNA, a 40% inhibition in ALDH2 activity, and a fourfold (P < 0.001) increase in circulating plasma acetaldehyde levels after ethanol (1 g/kg) administration. Administration of ASO-9 to rats by osmotic pumps led to an aversion (-61%, P < 0.02) to ethanol. These studies provide a proof of principle that specific inhibition of gene expression can be used to mimic the protective effects afforded by the ALDH2-2 phenotype.
Garver
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Department of Pathology, Anatomy and Cell Biology, and the Alcohol Research Center, Thomas Jefferson University, Philadelphia PA 19107, USA. emg002@jefferson.edu
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Cao
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Aini
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Zhou
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Israel
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D000079:Acetaldehyde / Q000097:blood; D000428:Alcohol Drinking / Q000097:blood / Q000235:genetics*; D000444:Aldehyde Dehydrogenase / Q000235:genetics* / Q000378:metabolism*; D000071396:Aldehyde Dehydrogenase, Mitochondrial; D000818:Animals; D001208:Asia / Q000208:ethnology; D044466:Asian People / Q000235:genetics*; D003513:Cycloheximide / Q000494:pharmacology; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects*; D005969:Glutamate Dehydrogenase / Q000378:metabolism; D006801:Humans; D008099:Liver / Q000201:enzymology; D008114:Liver Neoplasms, Experimental; D008297:Male; D008930:Mitochondria, Liver / Q000201:enzymology*; D020319:Oligodeoxyribonucleotides, Antisense / Q000494:pharmacology*; D010641:Phenotype; D012333:RNA, Messenger / Q000235:genetics; D051381:Rats; D011917:Rats, Inbred Lew; D017207:Rats, Sprague-Dawley; D011994:Recombinant Proteins / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014158:Transcription, Genetic / Q000187:drug effects; D014162:Transfection; D014407:Tumor Cells, Cultured; D014868:Water Deprivation
D020319:Oligodeoxyribonucleotides, Antisense; D012333:RNA, Messenger; D011994:Recombinant Proteins; D003513:Cycloheximide; C096127:ALDH2 protein, human; D000444:Aldehyde Dehydrogenase; D000071396:Aldehyde Dehydrogenase, Mitochondrial; D005969:Glutamate Dehydrogenase; D000079:Acetaldehyde