Drug-Induced Alterations of Mitochondrial DNA Homeostasis in Steatotic and Nonsteatotic HepaRG Cells

Dounia Le Guillou (1) , Simon Bucher (1) , Karima Begriche (1) , Delphine Hoët (2) , Anne Lombès (3) , Gilles Labbe (2) , Bernard Fromenty (1)

1 NuMeCan - Nutrition, Métabolismes et Cancer
2 SANOFI Recherche
3 IC UM3 (UMR 8104 / U1016) - Institut Cochin

Although mitochondriotoxicity plays a major role in drug-induced hepatotoxicity, alteration of mitochondrial DNA (mtDNA) homeostasis has been described only with a few drugs. Because it requires long drug exposure, this mechanism of toxicity cannot be detected with investigations performed in isolated liver mitochondria or cultured cells exposed to drugs for several hours or a few days. Thus, a first aim of this study was to determine whether a 2-week treatment with nine hepatotoxic drugs could affect mtDNA homeostasis in HepaRG cells. Previous investigations with these drugs showed rapid toxicity on oxidative phosphorylation but did not address the possibility of delayed toxicity secondary to mtDNA homeostasis impairment. The maximal concentration used for each drug induced about 10% cytotoxicity. Two other drugs, zalcitabine and linezolid, were used as positive controls for their respective effects on mtDNA replication and translation. Another goal was to determine whether drug-induced mitochondriotoxicity could be modulated by lipid overload mimicking nonalcoholic fatty liver. Among the nine drugs, imipramine and ritonavir induced mitochondrial effects suggesting alteration of mtDNA translation. Ritonavir toxicity was stronger in nonsteatotic cells. None of the nine drugs decreased mtDNA levels. However, increased mtDNA was observed with five drugs, especially in nonsteatotic cells. The mtDNA levels could not be correlated with the expression of key factors involved in mitochondrial biogenesis, such as peroxisome proliferator-activated receptor-γ coactivator 1α (PGC1α), PGC1β, and AMP-activated protein kinase α-subunit. Hence, drug-induced impairment of mtDNA translation might not be rare, and increased mtDNA levels could be a frequent adaptive response to slight energy shortage. Nevertheless, this adaptation could be impaired by lipid overload.

Mots clés: fatty liver-disease; respiratory-chain; hepg2 cells; injury; dysfunction; biogenesis; mechanisms; inhibition; ritonavir; toxicity

Domaines: Sciences pharmaceutiques; Toxicologie

Dounia Le Guillou, Simon Bucher, Karima Begriche, Delphine Hoët, Anne Lombès, et al.. Drug-Induced Alterations of Mitochondrial DNA Homeostasis in Steatotic and Nonsteatotic HepaRG Cells. Journal of Pharmacology and Experimental Therapeutics, 2018, 365 (3), pp.711-726. ⟨10.1124/jpet.117.246751⟩. ⟨hal-01808097⟩