N-Homocysteinylation of HMGB1/2 Promotes Corpus Cavernosum Endothelial Senescence in Erectile Dysfunction

Int J Biol Sci. 2025 Oct 20;21(15):6723-6744. doi: 10.7150/ijbs.119514. eCollection 2025.

Abstract

Homocysteine (Hcy) is an age-related risk factor for erectile dysfunction (ED), with enhanced vascular toxicity in middle-aged and elderly individuals. However, folate-based Hcy-lowering therapies have shown limited efficacy, necessitating a reevaluation of its age-dependent pathogenic mechanism. Here, we demonstrate that senescent endothelial cells exhibit heightened responsiveness of methionyl-tRNA synthetase 1 (MARS1) to Hcy, promoting the production of homocysteine thiolactone (HTL) and widespread N-homocysteinylation (K-Hcy) of proteins. K-Hcy, rather than acetylation, drives cytoplasmic translocation and extracellular release of high mobility group box proteins 1 and 2 (HMGB1/2), amplifying the senescence-associated secretory phenotype (SASP). Competitive inhibition of MARS1 with N-acetylcysteine (NAC) attenuates endothelial senescence and improves erectile function in middle-aged individuals with hyperhomocysteinemia by reducing HTL, rather than Hcy itself, while synergizing with tadalafil. Collectively, our findings highlight the pivotal role of the age-dependent MARS1-HTL axis in the pathogenesis of homocysteine-induced ED, offering a promising therapeutic strategy for ED in the aging population.

Keywords: High mobility group box 1 and 2; N-homocysteinylation; cellular senescence; erectile dysfunction; methionyl-tRNA synthetase 1.

MeSH terms

  • Animals
  • Cellular Senescence / physiology
  • Endothelial Cells / metabolism
  • Erectile Dysfunction* / metabolism
  • HMGB1 Protein* / metabolism
  • Homocysteine* / analogs & derivatives
  • Homocysteine* / metabolism
  • Humans
  • Hyperhomocysteinemia / metabolism
  • Male
  • Middle Aged
  • Penis* / metabolism

Substances

  • Homocysteine
  • HMGB1 Protein
  • homocysteine thiolactone