Involvement of BRD4 in Alcoholic Liver Injury: Autophagy Modulation via Regulation of the SIRT1/Beclin1 Axis

Lab Invest. 2024 Nov;104(11):102134. doi: 10.1016/j.labinv.2024.102134. Epub 2024 Sep 20.

Abstract

Alcoholic liver disease (ALD) caused by chronic alcohol abuse involves complex processes from steatosis to fibrosis, cirrhosis, and hepatocellular carcinoma, posing a global health issue. Bromodomain protein 4 (BRD4) typically serves as a "reader" modulating the functions of transcription factors involved in various biological processes and disease progression. However, the specific mechanisms underlying alcoholic liver injury remain unclear. In this study, we detected aberrant BRD4 expression in the alcohol-induced ALD mouse model of chronic and binge ethanol feeding developed by the National Institute on Alcohol Abuse and Alcoholism, consistent with the in vitro results in Aml-12 mouse hepatocytes. Blocking and inhibiting BRD4 restored the impaired autophagic flux and lysosomal functions in alcohol-treated Aml-12 cells, whereas BRD4 overexpression reduced the expression levels of autophagy marker and lysosomal genes. Furthermore, mouse BRD4 knockdown, mediated by a short hairpin RNA carried by the adeno-associated virus serotype 8, significantly attenuated the alcohol-induced hepatocyte damage, including lipid deposition and inflammatory cell infiltration. Mechanistically, BRD4 overexpression in alcoholic liver injury inhibited the expression of sirtuin (SIRT)1 in Aml-12 cells. Chromatin immunoprecipitation and dual-luciferase reporter assays revealed that BRD4 functions as a transcription factor and suppressor, actively binding to the SIRT1 promoter region and inhibiting its transcription. SIRT1 activated autophagy, which was suppressed in alcoholic liver injury via Beclin1 deacetylation. In conclusion, our study revealed that BRD4 negatively regulated the SIRT1/Beclin1 axis and that its deficiency alleviated alcohol-induced liver injury in mice, thus providing a new strategy for ALD treatment.

Keywords: BRD4; SIRT1; alcoholic liver injury; autophagy; epigenetics.

MeSH terms

  • Animals
  • Autophagy*
  • Beclin-1* / metabolism
  • Bromodomain Containing Proteins
  • Cell Line
  • Hepatocytes / metabolism
  • Liver / metabolism
  • Liver / pathology
  • Liver Diseases, Alcoholic* / metabolism
  • Male
  • Mice
  • Mice, Inbred C57BL*
  • Nuclear Proteins / genetics
  • Nuclear Proteins / metabolism
  • Sirtuin 1* / genetics
  • Sirtuin 1* / metabolism
  • Transcription Factors* / genetics
  • Transcription Factors* / metabolism

Substances

  • Sirtuin 1
  • Brd4 protein, mouse
  • Transcription Factors
  • Beclin-1
  • Sirt1 protein, mouse
  • Nuclear Proteins
  • Becn1 protein, mouse
  • Bromodomain Containing Proteins