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BMSC-derived exosomes ameliorate sulfur mustard-induced acute lung injury by regulating the GPRC5A–YAP axis

Abstract

Sulfur mustard (SM) is a highly toxic chemical warfare agent that causes acute lung injury (ALI) and/or acute respiratory distress syndrome (ARDS). There are no effective therapeutic treatments or antidotes available currently to counteract its toxic effects. Our previous study shows that bone marrow-derived mesenchymal stromal cells (BMSCs) could exert therapeutic effects against SM-induced lung injury. In this study, we explored the therapeutic potential of BMSC-derived exosomes (BMSC-Exs) against ALI and the underlying mechanisms. ALI was induced in mice by injection of SM (30 mg/kg, sc) at their medial and dorsal surfaces. BMSC-Exs (20 μg/kg in 200 μL PBS, iv) were injected for a 5-day period after SM exposure. We showed that BMSC-Exs administration caused a protective effect against pulmonary edema. Using a lung epithelial cell barrier model, BMSC-Exs (10, 20, 40 μg) dose-dependently inhibited SM-induced cell apoptosis and promoted the recovery of epithelial barrier function by facilitating the expression and relocalization of junction proteins (E-cadherin, claudin-1, occludin, and ZO-1). We further demonstrated that BMSC-Exs protected against apoptosis and promoted the restoration of barrier function against SM through upregulating G protein-coupled receptor family C group 5 type A (GPRC5A), a retinoic acid target gene predominately expressed in the epithelial cells of the lung. Knockdown of GPRC5A reduced the antiapoptotic and barrier regeneration abilities of BMSC-Exs and diminished their therapeutic effects in vitro and in vivo. BMSC-Exs-caused upregulation of GPRC5A promoted the expression of Bcl-2 and junction proteins via regulating the YAP pathway. In summary, BMSC-Exs treatment exerts protective effects against SM-induced ALI by promoting alveolar epithelial barrier repair and may be an alternative approach to stem cell-based therapy.

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Fig. 1: BMSC-Exs ameliorated SM-induced acute lung injury.
Fig. 2: BMSC-Exs reduced SM-injured lung epithelial cell apoptosis.
Fig. 3: BMSC-Exs alleviated SM-induced epithelial barrier damage.
Fig. 4: BMSC-Exs protected lung epithelial cells from apoptosis and epithelial barrier damage by regulating GPRC5A in MLE-12 cells.
Fig. 5: GPRC5A knockout attenuated the therapeutic effects of BMSC-Exs on SM-induced ALI in mice.
Fig. 6: BMSC-Exs promoted the repair of lung epithelial cells by activating the YAP pathway via GPRC5A.

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Acknowledgements

We are grateful to You-heng Wei (State Key Laboratory of Genetic Engineering, Institute of Genetics, Fudan University, Shanghai, China) for providing technical assistance. This work was funded by the National Natural Science Foundation of China (81671858 and 81871521) and the Natural Science Foundation of Shanghai (20ZR1470300).

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GCM, CCG, and ZW performed the experiments, collected data, and prepared the manuscript. KX, QQX, and YL conceived the study and wrote the main manuscript. GCM, ZPP, and MXS checked the data and performed data analysis. XWH and WQM participated in the design of the study and helped draft the manuscript. JFC and QQX helped perform data analysis and prepare the images. All authors read and approved the final manuscript.

Corresponding authors

Correspondence to Ying Lu or Qing-qiang Xu or Kai Xiao.

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The authors declare no competing interests.

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Mao, Gc., Gong, Cc., Wang, Z. et al. BMSC-derived exosomes ameliorate sulfur mustard-induced acute lung injury by regulating the GPRC5A–YAP axis. Acta Pharmacol Sin (2021). https://doi.org/10.1038/s41401-021-00625-4

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Keywords

  • acute lung injury
  • sulfur mustard
  • BMSC-derived exosomes
  • alveolar epithelial barrier
  • GPRC5A
  • YAP

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