Saltar al contenido
Merck

Inhibiting cytosolic translation and autophagy improves health in mitochondrial disease.

Human molecular genetics (2015-06-05)
Min Peng, Julian Ostrovsky, Young Joon Kwon, Erzsebet Polyak, Joseph Licata, Mai Tsukikawa, Eric Marty, Jeffrey Thomas, Carolyn A Felix, Rui Xiao, Zhe Zhang, David L Gasser, Yair Argon, Marni J Falk
RESUMEN

Mitochondrial respiratory chain (RC) disease therapies directed at intra-mitochondrial pathology are largely ineffective. Recognizing that RC dysfunction invokes pronounced extra-mitochondrial transcriptional adaptations, particularly involving dysregulated translation, we hypothesized that translational dysregulation is itself contributing to the pathophysiology of RC disease. Here, we investigated the activities, and effects from direct inhibition, of a central translational regulator (mTORC1) and its downstream biological processes in diverse genetic and pharmacological models of RC disease. Our data identify novel mechanisms underlying the cellular pathogenesis of RC dysfunction, including the combined induction of proteotoxic stress, the ER stress response and autophagy. mTORC1 inhibition with rapamycin partially ameliorated renal disease in B6.Pdss2(kd/kd) mice with complexes I-III/II-III deficiencies, improved viability and mitochondrial physiology in gas-1(fc21) nematodes with complex I deficiency, and rescued viability across a variety of RC-inhibited human cells. Even more effective was probucol, a PPAR-activating anti-lipid drug that we show also inhibits mTORC1. However, directly inhibiting mTORC1-regulated downstream activities yielded the most pronounced and sustained benefit. Partial inhibition of translation by cycloheximide, or of autophagy by lithium chloride, rescued viability, preserved cellular respiratory capacity and induced mitochondrial translation and biogenesis. Cycloheximide also ameliorated proteotoxic stress via a uniquely selective reduction of cytosolic protein translation. RNAseq-based transcriptome profiling of treatment effects in gas-1(fc21) mutants provide further evidence that these therapies effectively restored altered translation and autophagy pathways toward that of wild-type animals. Overall, partially inhibiting cytosolic translation and autophagy offer novel treatment strategies to improve health across the diverse array of human diseases whose pathogenesis involves RC dysfunction.

MATERIALES
Product Number
Marca
Descripción del producto

Sigma-Aldrich
Bicarbonato de sodio, powder, BioReagent, Molecular Biology, suitable for cell culture, suitable for insect cell culture
Sigma-Aldrich
Azul de tripano solution, 0.4%, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
Cicloheximida, from microbial, ≥94% (TLC)
Sigma-Aldrich
Cycloheximide solution, Ready-Made Solution, microbial, 100 mg/mL in DMSO, Suitable for cell culture
Sigma-Aldrich
Fluoruro de fenilmetansulfonilo, ≥98.5% (GC)
Sigma-Aldrich
Carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone, ≥98% (TLC), powder
Sigma-Aldrich
Rotenone, ≥95%
Sigma-Aldrich
Selenium, powder, −100 mesh, 99.99% trace metals basis
Sigma-Aldrich
Uridine, powder, BioReagent, suitable for cell culture
Sigma-Aldrich
Cycloheximide, ≥95% (HPLC)
Sigma-Aldrich
Uridine, ≥99%
Sigma-Aldrich
Fluoruro de fenilmetansulfonilo, ≥99.0% (T)
Sigma-Aldrich
Trypan Blue, powder, BioReagent, suitable for cell culture
Sigma-Aldrich
Selenium, powder, −100 mesh, ≥99.5% trace metals basis
Sigma-Aldrich
L-(−)-Glucose, ≥99%
Sigma-Aldrich
Bicarbonato de sodio, BioXtra, 99.5-100.5%
Sigma-Aldrich
Cicloheximida, Biotechnology Performance Certified
Sigma-Aldrich
Trypan Blue, ≥80% (HPLC), Dye content 60 %
Sigma-Aldrich
tert-Butyl acetoacetate, reagent grade, 98%
Sigma-Aldrich
Selenium, pellets, <5 mm particle size, ≥99.999% trace metals basis
Sigma-Aldrich
Selenium, pellets, <5 mm, ≥99.99% trace metals basis
Sigma-Aldrich
Uridine, BioUltra, ≥99%
Sigma-Aldrich
Sodium bicarbonate-12C, 99.9 atom % 12C
Selenium, foil, 25x25mm, thickness 3mm, 99.95%
Selenium, pellets, < 5mm, ≥99.999%