Modulating in vitro lung fibroblast activation via senolysis of senescent human alveolar epithelial cells

Aging (Albany NY). 2024 Jun 29;16(13):10694-10723. doi: 10.18632/aging.205994. Epub 2024 Jun 29.

Abstract

Idiopathic pulmonary fibrosis (IPF) is an age-related disease with poor prognosis and limited therapeutic options. Activation of lung fibroblasts and differentiation to myofibroblasts are the principal effectors of disease pathology, but damage and senescence of alveolar epithelial cells, specifically type II (ATII) cells, has recently been identified as a potential trigger event for the progressive disease cycle. Targeting ATII senescence and the senescence-associated secretory phenotype (SASP) is an attractive therapeutic strategy; however, translatable primary human cell models that enable mechanistic studies and drug development are lacking. Here, we describe a novel system of conditioned medium (CM) transfer from bleomycin-induced senescent primary alveolar epithelial cells (AEC) onto normal human lung fibroblasts (NHLF) that demonstrates an enhanced fibrotic transcriptional and secretory phenotype compared to non-senescent AEC CM treatment or direct bleomycin damage of the NHLFs. In this system, the bleomycin-treated AECs exhibit classical hallmarks of cellular senescence, including SASP and a gene expression profile that resembles aberrant epithelial cells of the IPF lung. Fibroblast activation by CM transfer is attenuated by pre-treatment of senescent AECs with the senolytic Navitoclax and AD80, but not with the standard of care agent Nintedanib or senomorphic JAK-targeting drugs (e.g., ABT-317, ruxolitinib). This model provides a relevant human system for profiling novel senescence-targeting therapeutics for IPF drug development.

Keywords: SASP; alveolar epithelial cell; cellular senescence; fibrosis; senolytic; senomorphic.

MeSH terms

  • Alveolar Epithelial Cells* / drug effects
  • Alveolar Epithelial Cells* / metabolism
  • Alveolar Epithelial Cells* / pathology
  • Aniline Compounds
  • Bleomycin* / pharmacology
  • Bleomycin* / toxicity
  • Cells, Cultured
  • Cellular Senescence* / drug effects
  • Culture Media, Conditioned / pharmacology
  • Fibroblasts* / drug effects
  • Fibroblasts* / metabolism
  • Humans
  • Idiopathic Pulmonary Fibrosis* / metabolism
  • Idiopathic Pulmonary Fibrosis* / pathology
  • Indoles / pharmacology
  • Lung / cytology
  • Lung / drug effects
  • Lung / pathology
  • Nitriles / pharmacology
  • Pyrazoles / pharmacology
  • Pyrimidines / pharmacology
  • Senescence-Associated Secretory Phenotype / drug effects
  • Senotherapeutics / pharmacology
  • Sulfonamides / pharmacology

Substances

  • Bleomycin
  • Culture Media, Conditioned
  • navitoclax
  • Indoles
  • Sulfonamides
  • Senotherapeutics
  • Pyrimidines
  • nintedanib
  • Pyrazoles
  • Nitriles
  • ruxolitinib
  • Aniline Compounds