title: Statins induce apoptosis in rat and human myotube cultures by inhibiting protein geranylgeranylation but not ubiquinone.
abstract: Statins are widely used to treat lipid disorders. These drugs are safe and well tolerated; however, in <1% of patients, myopathy and/or rhabdomyolysis can develop. To better understand the mechanism of statin-induced myopathy, we examined the ability of structurally distinct statins to induce apoptosis in an optimized rat myotube model. Compound A (a lactone) and Cerivastatin (an open acid) induced apoptosis, as measured by TUNEL and active caspase 3 staining, in a concentration- and time-dependent manner. In contrast, an epimer of Compound A (Compound B) exhibited a much weaker apoptotic response. Statin-induced apoptosis was completely prevented by mevalonate or geranylgeraniol, but not by farnesol. Zaragozic acid A, a squalene synthase inhibitor, caused no apoptosis on its own and had no effect on Compound-A-induced myotoxicity, suggesting the apoptosis was not a result of cholesterol synthesis inhibition. The geranylgeranyl transferase inhibitors GGTI-2133 and GGTI-2147 caused apoptosis in myotubes; the farnesyl transferase inhibitor FTI-277 exhibited a much weaker effect. In addition, the prenylation of rap1a, a geranylgeranylated protein, was inhibited by Compound A in myotubes at concentrations that induced apoptosis. A similar statin-induced apoptosis profile was seen in human myotube cultures but primary rat hepatocytes were about 200-fold more resistant to statin-induced apoptosis. Although the statin-induced hepatotoxicity could be attenuated with mevalonate, no effect was found with either geranylgeraniol or farnesol. In studies assessing ubiquinone levels after statin treatment in rat and human myotubes, there was no correlation between ubiquinone levels and apoptosis. Taken together, these observations suggest that statins cause apoptosis in myotube cultures in part by inhibiting the geranylgeranylation of proteins, but not by suppressing ubiquinone concentration. Furthermore, the data from primary hepatocytes suggests a cell-type differential sensitivity to statin-induced toxicity.
authors.affiliation: Department of Safety Assessment, Merck Research Laboratories, West Point, PA 19486, USA. Timothy_Johnson@merck.com
authors.forename: Timothy E
authors.initials: TE
authors.lastname: Johnson
authors.forename: Xiaohua
authors.initials: X
authors.lastname: Zhang
authors.forename: Kimberly B
authors.initials: KB
authors.lastname: Bleicher
authors.forename: Gary
authors.initials: G
authors.lastname: Dysart
authors.forename: Amy F
authors.initials: AF
authors.lastname: Loughlin
authors.forename: William H
authors.initials: WH
authors.lastname: Schaefer
authors.forename: Diane R
authors.initials: DR
authors.lastname: Umbenhauer
mesh_terms: D019883:Alkyl and Aryl Transferases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D002469:Cell Separation; D002478:Cells, Cultured; D003402:Creatine Kinase / Q000378:metabolism; D006801:Humans; D019161:Hydroxymethylglutaryl-CoA Reductase Inhibitors / Q000494:pharmacology*; D020287:In Situ Nick-End Labeling; D008297:Male; D018485:Muscle Fibers, Skeletal / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D017368:Protein Prenylation; D011506:Proteins / Q000378:metabolism*; D012333:RNA, Messenger / Q000032:analysis / Q000096:biosynthesis; D051381:Rats; D017207:Rats, Sprague-Dawley; D014451:Ubiquinone / Q000378:metabolism*; D020827:rap1 GTP-Binding Proteins / Q000378:metabolism
chemical_list: D019161:Hydroxymethylglutaryl-CoA Reductase Inhibitors; D011506:Proteins; D012333:RNA, Messenger; D014451:Ubiquinone; D019883:Alkyl and Aryl Transferases; C083291:geranylgeranyltransferase type-I; D003402:Creatine Kinase; D020827:rap1 GTP-Binding Proteins