A cGAS-dependent response links DNA damage and senescence in alveolar epithelial cells: A potential drug target in IPF

Michael Schuliga*, Amama Kanwal, Jane Read, Kaj Erik Cornelis Blokland, Janette K Burgess, Cecilia M Prêle, Steven E Mutsaers, Christopher Grainge, Claire Thomson, Allen James, Nathan W Bartlett, Darryl A Knight

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

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Abstract

Alveolar epithelial cell (AEC) senescence is implicated in the pathogenesis of idiopathic pulmonary fibrosis (IPF). Mitochondrial dysfunction including release of mitochondrial DNA (mtDNA) is a feature of senescence, which led us to investigate the role of the DNA-sensing GMP-AMP synthase (cGAS) in IPF, with a focus on AEC senescence. cGAS expression in fibrotic tissue from lungs of IPF patients was detected within cells immunoreactive for epithelial cell adhesion molecule (EpCAM) and p21, epithelial and senescence markers respectively. Submerged primary cultures of AECs isolated from lung tissue of IPF patients (IPF-AECs, n=5) exhibited higher baseline senescence than AECs from control donors (Ctrl-AECs, n=5-7), as assessed by increased nuclear histone 2AXγ phosphorylation, p21 mRNA and expression of senescence-associated secretory phenotype (SASP) cytokines. Pharmacological cGAS inhibition using RU.521 diminished IPF-AEC senescence in culture and attenuated induction of Ctrl-AEC senescence following etoposide-induced DNA damage. Short interfering RNA (siRNA) knockdown of cGAS also attenuated etoposide-induced senescence of the AEC line, A549. Higher levels of mtDNA were detected in the cytosol and culture supernatants of primary IPF- and etoposide-treated Ctrl-AECs when compared to Ctrl-AECs at baseline. Furthermore, ectopic mtDNA augmented cGAS-dependent senescence of Ctrl-AECs, whereas DNAse I treatment diminished IPF-AEC senescence. This study provides evidence that a self-DNA driven, cGAS-dependent response augments AEC senescence, identifying cGAS as a potential therapeutic target for IPF.

Original languageEnglish
Pages (from-to)L859-L871
Number of pages41
JournalAmerican Journal of Physiology - Lung Cellular and Molecular Physiology
Volume321
Issue number5
DOIs
Publication statusPublished - Nov-2021

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