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Exosomal MiR-381 from M2-polarized macrophages attenuates urethral fibroblasts activation through YAP/GLS1-regulated glutaminolysis

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Abstract

Objective and design

Post-traumatic urethral stricture is a clinical challenge for both patients and clinicians. Targeting glutamine metabolism to suppress excessive activation of urethral fibroblasts (UFBs) is assumed to be a potent and attractive strategy for preventing urethral scarring and stricture.

Material or subjects

In cellular experiments, we explored whether glutaminolysis meets the bioenergetic and biosynthetic demands of quiescent UFBs converted into myofibroblasts. At the same time, we examined the specific effects of M2-polarized macrophages on glutaminolysis and activation of UFBs, as well as the mechanism of intercellular signaling. In addition, findings were further verified in vivo in New Zealand rabbits.

Results

It revealed that glutamine deprivation or knockdown of glutaminase 1 (GLS1) significantly inhibited UFB activation, proliferation, biosynthesis, and energy metabolism; however, these effects were rescued by cell-permeable dimethyl α-ketoglutarate. Moreover, we found that exosomal miR-381 derived from M2-polarized macrophages could be ingested by UFBs and inhibited GLS1-dependent glutaminolysis, thereby preventing excessive activation of UFBs. Mechanistically, miR-381 directly targets the 3′UTR of Yes-associated protein (YAP) mRNA to reduce its stability at the transcriptional level, ultimately downregulating expression of YAP, and GLS1. In vivo experiments revealed that treatment with either verteporfin or exosomes derived from M2-polarized macrophages significantly reduced urethral stricture in New Zealand rabbits after urethral trauma.

Conclusion

Collectively, this study demonstrates that exosomal miR-381 from M2-polarized macrophages reduces myofibroblast formation of UFBs and urethral scarring and stricture by inhibiting YAP/GLS1-dependent glutaminolysis.

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Data availability

Not applicable.

Abbreviations

UFBs:

Urethral fibroblasts

GLS1:

Glutaminase 1

YAP:

Yes-associated protein

PTUS:

Post‐traumatic urethral stricture

α‐KG:

α‐Ketoglutarate

TGF-β1:

Transforming growth factor β1

qRT-PCR:

Quantitative real-time polymerase chain reaction

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Acknowledgements

Not applicable.

Funding

This study was supported by National Natural Science Foundation of China (Grant number: 81970589), Foundation of Fujian Provincial Department of Finance (Grant number: 2019B030), and the National Natural Science Foundation of Fujian Provincial (Grant number: 2020J01989).

Author information

Author notes

  1. Ye-Hui Chen, Yi-Cheng Xu, Ting-Ting Lin and Hang Chen have equally contributed to this work.

Authors and Affiliations

  1. Department of Urology, Urology Research Institute, The First Affiliated Hospital, Fujian Medical University, Fuzhou, 350005, China

    Ye-Hui Chen, Yi-Cheng Xu, Ting-Ting Lin, Hang Chen, Ru-Nan Dong, Zhi-Bin Ke, Jia-Yin Chen, Yong Wei, Qing-Shui Zheng, Xue-Yi Xue & Ning Xu

  2. Department of Urology, National Regional Medical Center, Binhai Campus of the First Affiliated Hospital, Fujian Medical University, Fuzhou, 350212, China

    Ye-Hui Chen, Yi-Cheng Xu, Ting-Ting Lin, Hang Chen, Ru-Nan Dong, Zhi-Bin Ke, Jia-Yin Chen, Yong Wei, Qing-Shui Zheng, Xue-Yi Xue & Ning Xu

  3. Fujian Key Laboratory of Precision Medicine for Cancer, The First Affiliated Hospital, Fujian Medical University, 20 Chazhong Road, Fuzhou, 350005, China

    Xue-Yi Xue & Ning Xu

  4. Department of Ultrasonography, The First Affiliated Hospital, Fujian Medical University, Fuzhou, 350005, China

    Feng-Ping Cai

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Contributions

Y-HC, Y-CX, T-TL, and HC wrote the original draft. X-YX and NX reviewed and edited the original draft. J-YC and Y-HC contributed to the methodology. T-TL and F-PC did the investigation. Z-BK and HC contributed to formal analysis. R-ND and Y-CX curated the data. Y-HC contributed to the conceptualization and the visualization. YW and Q-SZ supervised the project. NX and X-YX administrated the project.

Corresponding authors

Correspondence to Xue-Yi Xue or Ning Xu.

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Competing interests

The authors declare no competing interests.

Conflict of interest

All authors declared that there were no conflicts of interest.

Ethical approval

All experiments with primary cells were in accordance with the Declaration of Helsinki and were approved by the Ethics Committee of the First Affiliated Hospital of Fujian Medical University. All donors of primary cells have provided informed consents. All experiments were performed according to the written confirmation that this study was in accordance with relevant institutional guidelines and regulations of Fujian Medical University and national guidelines and regulations of China. All animal experiments were approved by the Experimental Animal Ethics Committee of Fujian Medical University.

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Chen, YH., Xu, YC., Lin, TT. et al. Exosomal MiR-381 from M2-polarized macrophages attenuates urethral fibroblasts activation through YAP/GLS1-regulated glutaminolysis. Inflamm. Res. 72, 1359–1373 (2023). https://doi.org/10.1007/s00011-023-01735-x

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