Alveolar macrophage-derived exosomal tRF-22-8BWS7K092 activates Hippo signaling pathway to induce ferroptosis in acute lung injury.

Background: Alveolar macrophages (AMs) play a demonstrative role in acute lung injury (ALI). Exosomes act as signaling molecules to regulate cell-to-cell communication by releasing RNAs. Transfer RNA-derived fragments (tRFs) possess potential functions in multiple diseases through ferroptosis. The present study aims to reveal the role of AM-derived exosomal tRFs in ALI and to identify the relationship to ferroptosis.

Methods: ALI mice model was established by lipopolysaccharide (LPS) induction. RNA sequencing was performed to identify the tRFs profile in bronchoalveolar lavage fluid (BALF) exosomes of ALI mice. After interfering with the expression of candidate tRFs in AMs or alveolar epithelial cells (MLE-12), the effect of oxidative stress and expression of ferroptosis-related proteins were detected.

Results: Exosomes isolated from BALF of ALI mice were dominated by a macrophage immunophenotype. RNA-sequencing identified 4 up- and 10 down-regulated differentially expressed tRFs (DEtRFs), among which tRF-22-8BWS7K092 expression was significantly increased in LPS-induced macrophage-derived exosomes (LPS-exo). Hippo signaling pathway was the most significantly enriched KEGG pathways for DEtRFs. LPS-exo inhibited cell viability and the expression of GPX4 and FTH1, and enhanced oxidative stress in MLE-12 cells. Ferroptosis inhibitor reversed the inhibition of LPS-exo on cell viability and tRF-22-8BWS7K092 inhibitor rescued above effect of LPS-exo on MLE-12 cells. Besides, tRF-22-8BWS7K092 could activate Hippo signaling pathway by binding Wnt5B, inducing ferroptosis in MLE-12 cells.

Conclusion: BALF exosomes of ALI mice were mainly derived from AMs. AM-derived exosomal tRF-22-8BWS7K092 activates the Hippo signaling pathway to induce ferroptosis, thus contributing to the pathogenesis of ALI.