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Circulating cell-free mtDNA release is associated with the activation of cGAS-STING pathway and inflammation in mitochondrial diseases

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Abstract

Background

There is increasing evidence for the role of inflammation in the pathogenesis of mitochondrial diseases (MDs). However, the mechanisms underlying mutation-induced inflammation in MD remain elusive. Our previous study suggested that mitophagy is impaired in the skeletal muscle of those with MD, likely causing mitochondrial DNA (mtDNA) release and thereby triggering inflammation. We here aimed to decipher the role of the cGAS-STING pathway in inflammatory process in MDs.

Methods

We investigated the levels of circulating cell-free mtDNA (ccf-mtDNA) in the serum of 104 patients with MDs. Immunofluorescence was performed in skeletal muscles in MDs and control. Biochemical analysis of muscle biopsies was conducted with western blot to detect cGAS, STING, TBK1, IRF3 and phosphorylated IRF3 (p-IRF3). RT-qPCR was performed to detect the downstream genes of type I interferon in skeletal muscles. Furthermore, a protein microarray was used to examine the cytokine levels in the serum of patients with MDs.

Results

We found that ccf-mtDNA levels were significantly increased in those with MDs compared to the controls. Consistently, the immunofluorescent results showed that cytosolic dsDNA levels were increased in the muscle samples of MD patients. Biochemical analysis of muscle biopsies showed that cGAS, IRF3, and TBK1 protein levels were significantly increased in those with MDs, indicating that there was activation of the cGAS-STING pathway. RT-qPCR showed that downstream genes of type I interferon were upregulated in muscle samples of MDs. Protein microarray results showed that a total of six cytokines associated with the cGAS-STING pathway were significantly increased in MD patients (fold change > 1.2, p value < 0.05).

Conclusions

These findings suggest that increases in ccf-mtDNA levels is associated with the activation of the cGAS-STING pathway, thereby triggering inflammation in MDs.

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Acknowledgements

We appreciated the cooperation of the patients and their families.

Funding

The work was funded by the National Natural Science Foundation of China (No. 81571219, 82071409, 82171846 and U20A20356). Capital's Funds for Health Improvement and Research (2022-4-40716).

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Author notes

  1. Xutong Zhao and Meng Yu contributed equally as co-first authors.

Authors and Affiliations

  1. Department of Neurology, Peking University First Hospital, #8 Xishiku St, Xicheng District, Beijing, 100034, China

    Xutong Zhao, Meng Yu, Yawen Zhao, Yiming Zheng, Lingchao Meng, Kang Du, Zhiying Xie, He Lv, Wei Zhang, Jing Liu, Qingqing Wang, Yun Yuan, Zhaoxia Wang & Jianwen Deng

  2. Beijing Key Laboratory of Neurovascular Disease Discovery, Beijing, 100034, China

    Zhaoxia Wang & Jianwen Deng

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  1. Xutong Zhao
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Contributions

ZXW and JWD conceived the idea, designed studies and supervised the project; XTZ and MY designed and carried out experiments, analyzed data; MY, YWZ, YMZ, ZYX, and LCM collected the patient; QQW, JL, HL, WZ, YY contributed to the clinical diagnosis and biopsy of MD patients; XTZ, ZXW and JWD wrote and edited the manuscript. All authors read and approved the final manuscript.

Corresponding authors

Correspondence to Zhaoxia Wang or Jianwen Deng.

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The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

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Zhao, X., Yu, M., Zhao, Y. et al. Circulating cell-free mtDNA release is associated with the activation of cGAS-STING pathway and inflammation in mitochondrial diseases. J Neurol 269, 4985–4996 (2022). https://doi.org/10.1007/s00415-022-11146-3

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