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Area of Science:

  • Ophthalmology
  • Molecular Biology
  • Cell Biology

Background:

  • Corneal neovascularization (CoNV) is a pathological process involving new blood vessel growth in the cornea.
  • Understanding the molecular regulators of CoNV is crucial for developing effective treatments.
  • Runt-related transcription factor 1 (RUNX1) has emerged as a potential key player in angiogenesis.

Purpose of the Study:

  • To investigate the regulatory role of RUNX1 in corneal neovascularization (CoNV).
  • To elucidate the underlying molecular mechanisms, including the involvement of P300 and autophagy.

Main Methods:

  • In vitro studies using VEGF-induced human umbilical vein endothelial cells (HUVECs) with RUNX1/P300 manipulation.
  • Assays for cell proliferation (EdU), migration (Transwell), and tube formation.
  • Chromatin immunoprecipitation (ChIP) and co-immunoprecipitation (Co-IP) to assess protein interactions.
  • Autophagic flux monitoring using RFP-GFP-LC3 reporters.
  • In vivo validation using a rat model of alkali burn-induced CoNV.

Main Results:

  • RUNX1 and P300 levels were elevated in CoNV and VEGF-induced HUVECs, correlating with inhibited autophagy.
  • P300 directly binds to the RUNX1 promoter, enhancing RUNX1 transcription.
  • RUNX1 overexpression counteracted the anti-angiogenic effects of P300 silencing.
  • RUNX1 knockdown attenuated VEGF-induced HUVEC proliferation, migration, and tube formation by activating autophagy.

Conclusions:

  • P300 enhances RUNX1 transcription by binding to its promoter.
  • RUNX1 plays a critical role in CoNV progression by regulating autophagy-related genes.
  • Targeting the RUNX1-P300-autophagy axis presents a potential therapeutic strategy for CoNV.