In vitro screening of cell bioenergetics to assess mitochondrial dysfunction in drug development

K. Tilmant (Lead / Corresponding author), H. Gerets, P. De Ron, E. Hanon, C. Bento-Pereira, F. A. Atienzar

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Drug-induced mitochondrial toxicity is considered as a common cellular mechanism that can induce a variety of organ toxicities. In the present manuscript, 17 in vitro mitochondrial toxic drugs, reported to induce Drug-Induced Liver Injury (DILI) and 6 non-mitochondrial toxic drugs (3 with DILI and 3 without DILI concern), were tested in HepG2 cells using a bioenergetics system. The 17 mitochondrial toxic drugs represent a wide variety of mitochondrial dysfunctions as well as DILI and include 4 pairs of drugs which are structurally related but associated with different DILI concerns in human. Cell bioenergetics were measured using the XF96e analyzer which simultaneous monitor oxygen consumption rate (OCR) and extracellular acidification rate (ECAR), indirect measurements of oxidative phosphorylation and glycolysis, respectively. OCR associated with ATP production, maximal respiration, proton leak and spare respiratory capacity, were also assessed. Duplicate experiments resulted in a sensitivity of 82% (14/17) and specificity of 83% (5/6). The addition of stressors improved specificity considerably. Cut-offs, statistics and rules are clearly discussed to facilitate the use of this assay for screening purposes. Overall, the authors consider that this assay should be part of the battery of safety screening assays at early stages of drug development.

Original languageEnglish
Pages (from-to)374-383
Number of pages10
JournalToxicology in Vitro
Volume52
Early online date17 Jul 2018
DOIs
Publication statusPublished - 1 Oct 2018

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Chemical and Drug Induced Liver Injury
Energy Metabolism
Screening
Poisons
Pharmaceutical Preparations
Liver
Oxygen Consumption
Assays
Manuscripts
Oxidative Phosphorylation
Hep G2 Cells
Glycolysis
Toxicity
In Vitro Techniques
Protons
Respiration
Adenosine Triphosphate
Oxygen
Safety
Acidification

Keywords

  • bioenergetics
  • HepG2
  • mitochondria
  • OCR/ECAR
  • screening
  • XF96e
  • Drug Evaluation, Preclinical/methods
  • Drug-Related Side Effects and Adverse Reactions
  • Humans
  • Energy Metabolism/drug effects
  • Mitochondria/drug effects
  • Hep G2 Cells

Cite this

Tilmant, K. ; Gerets, H. ; De Ron, P. ; Hanon, E. ; Bento-Pereira, C. ; Atienzar, F. A. / In vitro screening of cell bioenergetics to assess mitochondrial dysfunction in drug development. In: Toxicology in Vitro. 2018 ; Vol. 52. pp. 374-383.
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In vitro screening of cell bioenergetics to assess mitochondrial dysfunction in drug development. / Tilmant, K. (Lead / Corresponding author); Gerets, H.; De Ron, P.; Hanon, E.; Bento-Pereira, C.; Atienzar, F. A.

In: Toxicology in Vitro, Vol. 52, 01.10.2018, p. 374-383.

Research output: Contribution to journalArticle

TY - JOUR

T1 - In vitro screening of cell bioenergetics to assess mitochondrial dysfunction in drug development

AU - Tilmant, K.

AU - Gerets, H.

AU - De Ron, P.

AU - Hanon, E.

AU - Bento-Pereira, C.

AU - Atienzar, F. A.

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Y1 - 2018/10/1

N2 - Drug-induced mitochondrial toxicity is considered as a common cellular mechanism that can induce a variety of organ toxicities. In the present manuscript, 17 in vitro mitochondrial toxic drugs, reported to induce Drug-Induced Liver Injury (DILI) and 6 non-mitochondrial toxic drugs (3 with DILI and 3 without DILI concern), were tested in HepG2 cells using a bioenergetics system. The 17 mitochondrial toxic drugs represent a wide variety of mitochondrial dysfunctions as well as DILI and include 4 pairs of drugs which are structurally related but associated with different DILI concerns in human. Cell bioenergetics were measured using the XF96e analyzer which simultaneous monitor oxygen consumption rate (OCR) and extracellular acidification rate (ECAR), indirect measurements of oxidative phosphorylation and glycolysis, respectively. OCR associated with ATP production, maximal respiration, proton leak and spare respiratory capacity, were also assessed. Duplicate experiments resulted in a sensitivity of 82% (14/17) and specificity of 83% (5/6). The addition of stressors improved specificity considerably. Cut-offs, statistics and rules are clearly discussed to facilitate the use of this assay for screening purposes. Overall, the authors consider that this assay should be part of the battery of safety screening assays at early stages of drug development.

AB - Drug-induced mitochondrial toxicity is considered as a common cellular mechanism that can induce a variety of organ toxicities. In the present manuscript, 17 in vitro mitochondrial toxic drugs, reported to induce Drug-Induced Liver Injury (DILI) and 6 non-mitochondrial toxic drugs (3 with DILI and 3 without DILI concern), were tested in HepG2 cells using a bioenergetics system. The 17 mitochondrial toxic drugs represent a wide variety of mitochondrial dysfunctions as well as DILI and include 4 pairs of drugs which are structurally related but associated with different DILI concerns in human. Cell bioenergetics were measured using the XF96e analyzer which simultaneous monitor oxygen consumption rate (OCR) and extracellular acidification rate (ECAR), indirect measurements of oxidative phosphorylation and glycolysis, respectively. OCR associated with ATP production, maximal respiration, proton leak and spare respiratory capacity, were also assessed. Duplicate experiments resulted in a sensitivity of 82% (14/17) and specificity of 83% (5/6). The addition of stressors improved specificity considerably. Cut-offs, statistics and rules are clearly discussed to facilitate the use of this assay for screening purposes. Overall, the authors consider that this assay should be part of the battery of safety screening assays at early stages of drug development.

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SN - 0887-2333

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