Thymidine Analogues Suppress Autophagy and Adipogenesis in Cultured Adipocytes

Antimicrob Agents Chemother. 2013 Jan;57(1):543-51. doi: 10.1128/AAC.01560-12. Epub 2012 Nov 12.

Abstract

Lipoatrophy in HIV patients can result from prolonged exposure to thymidine analogues. Mitochondrial toxicity leading to dysregulated adipogenesis and increased cell death has been proposed as a leading factor in the etiology of peripheral fat loss. We hypothesized that thymidine analogues interfere with autophagy, a lysosomal degradation pathway, which is important for mitochondrial quality control, cellular survival, and adipogenesis. We assessed the effects of zidovudine (AZT), stavudine (d4T), and lamivudine (3TC) on autophagy in eukaryotic cells and adipocytes (3T3-F442A) by fluorescence microscopy and flow cytometry. The effects were compared to interventions with established genetic and pharmacological inhibitors of autophagy and correlated to assessments of cell viability, proliferation, and differentiation. AZT and d4T, but not 3TC, inhibited both constitutive and induced autophagic activity in adipocytes. This inhibition was associated with accumulation of dysfunctional mitochondria, increased reactive oxygen species (ROS) production, increased apoptosis, decreased proliferation, and impaired adipogenic conversion. Autophagy inhibition was dose and time dependent and detectable at therapeutic drug concentrations. Similar phenotypic changes were obtained when genetic or pharmacological inhibition of autophagy was employed. Our data suggest that thymidine analogues disturb adipocyte function through inhibition of autophagy. This novel mechanism potentially contributes to peripheral fat loss in HIV-infected patients.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adipocytes / cytology
  • Adipocytes / drug effects*
  • Adipocytes / metabolism
  • Adipogenesis / drug effects*
  • Androstadienes / pharmacology
  • Animals
  • Autophagy / drug effects*
  • Autophagy-Related Protein 5
  • Cell Differentiation / drug effects
  • Cell Line
  • Cell Proliferation / drug effects
  • Cell Survival / drug effects
  • Chromones / pharmacology
  • Flow Cytometry
  • Humans
  • Lamivudine / pharmacology*
  • Mice
  • Microscopy, Fluorescence
  • Microtubule-Associated Proteins / antagonists & inhibitors
  • Microtubule-Associated Proteins / genetics
  • Mitochondria / drug effects
  • Mitochondria / metabolism
  • Morpholines / pharmacology
  • Nocodazole / pharmacology
  • Phagosomes / drug effects
  • Phagosomes / metabolism
  • RNA, Small Interfering / genetics
  • Reverse Transcriptase Inhibitors / pharmacology*
  • Stavudine / pharmacology*
  • Wortmannin
  • Zidovudine / pharmacology*

Substances

  • ATG5 protein, human
  • Androstadienes
  • Autophagy-Related Protein 5
  • Chromones
  • Microtubule-Associated Proteins
  • Morpholines
  • RNA, Small Interfering
  • Reverse Transcriptase Inhibitors
  • Lamivudine
  • 2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one
  • Zidovudine
  • Stavudine
  • Nocodazole
  • Wortmannin