title: Mevalonate cascade regulation of airway mesenchymal cell autophagy and apoptosis: a dual role for p53.
abstract: Statins inhibit the proximal steps of cholesterol biosynthesis, and are linked to health benefits in various conditions, including cancer and lung disease. We have previously investigated apoptotic pathways triggered by statins in airway mesenchymal cells, and identified reduced prenylation of small GTPases as a primary effector mechanism leading to p53-mediated cell death. Here, we extend our studies of statin-induced cell death by assessing endpoints of both apoptosis and autophagy, and investigating their interplay and coincident regulation. Using primary cultured human airway smooth muscle (HASM) and human airway fibroblasts (HAF), autophagy, and autophagosome formation and flux were assessed by transmission electron microscopy, cytochemistry (lysosome number and co-localization with LC3) and immunoblotting (LC3 lipidation and Atg12-5 complex formation). Chemical inhibition of autophagy increased simvastatin-induced caspase activation and cell death. Similarly, Atg5 silencing with shRNA, thus preventing Atg5-12 complex formation, increased pro-apoptotic effects of simvastatin. Simvastatin concomitantly increased p53-dependent expression of p53 up-regulated modulator of apoptosis (PUMA), NOXA, and damage-regulated autophagy modulator (DRAM). Notably both mevalonate cascade inhibition-induced autophagy and apoptosis were p53 dependent: simvastatin increased nuclear p53 accumulation, and both cyclic pifithrin-α and p53 shRNAi partially inhibited NOXA, PUMA expression and caspase-3/7 cleavage (apoptosis) and DRAM expression, Atg5-12 complex formation, LC3 lipidation, and autophagosome formation (autophagy). Furthermore, the autophagy response is induced rapidly, significantly delaying apoptosis, suggesting the existence of a temporally coordinated p53 regulation network. These findings are relevant for the development of statin-based therapeutic approaches in obstructive airway disease.
authors.affiliation: Department of Physiology, University of Manitoba, Winnipeg, Manitoba, Canada.
authors.forename: Saeid
authors.initials: S
authors.lastname: Ghavami
authors.forename: Mark M
authors.initials: MM
authors.lastname: Mutawe
authors.forename: Pawan
authors.initials: P
authors.lastname: Sharma
authors.forename: Behzad
authors.initials: B
authors.lastname: Yeganeh
authors.forename: Karol D
authors.initials: KD
authors.lastname: McNeill
authors.forename: Thomas
authors.initials: T
authors.lastname: Klonisch
authors.forename: Helmut
authors.initials: H
authors.lastname: Unruh
authors.forename: Hessam H
authors.initials: HH
authors.lastname: Kashani
authors.forename: Dedmer
authors.initials: D
authors.lastname: Schaafsma
authors.forename: Marek
authors.initials: M
authors.lastname: Los
authors.forename: Andrew J
authors.initials: AJ
authors.lastname: Halayko
mesh_terms: D017209:Apoptosis / Q000187:drug effects*; D001343:Autophagy / Q000187:drug effects*; D002478:Cells, Cultured; D006801:Humans; D019161:Hydroxymethylglutaryl-CoA Reductase Inhibitors / Q000494:pharmacology; D008648:Mesoderm / Q000166:cytology* / Q000187:drug effects; D008798:Mevalonic Acid / Q000494:pharmacology*; D012137:Respiratory System / Q000166:cytology* / Q000187:drug effects; D019821:Simvastatin / Q000494:pharmacology; D016159:Tumor Suppressor Protein p53 / Q000502:physiology*
chemical_list: D019161:Hydroxymethylglutaryl-CoA Reductase Inhibitors; C495901:TP53 protein, human; D016159:Tumor Suppressor Protein p53; D019821:Simvastatin; D008798:Mevalonic Acid