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cGAS Deficiency Regulates the Phenotypic Polarization and Glycolysis of Microglia Through Lactylation in Hypoxic-Ischemic Encephalopathy Cell Model

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Abstract

Promoting the M2 phenotype polarization of microglia is of great significance in alleviating hypoxic-ischemic encephalopathy (HIE). The umbilical artery blood sample was collected to evaluate the expression of cGAS, and the aberrant expressed cGAS was verified in the oxygen glucose deprivation (OGD) microglia which was established to mimic HIE in vitro. Then the regulating role of cGAS on the transformation of microglia M2 phenotype polarization and glycolysis was investigated. Moreover, the lactylation of cGAS in OGD treated microglia was evaluated by western blot. cGAS was found to be highly expressed in umbilical artery blood of HIE group, and OGD treated microglia. OGD interference activated microglia into M1 phenotype by enhancing CD86 and suppressing CD206 levels; meanwhile, the microglia in OGD group highly expressed IL-1β, iNOS and TNF-α, and lowly expressed IL-4, IL-10, and Arg-1. Inhibition of cGAS promotes the transformation of microglia from M1 to M2 phenotype. Meanwhile, OGD increased ECAR and decreased OCR to regulate glycolysis, cGAS deficiency inhibits glycolysis in OGD treated microglia. Moreover, the pan lysine lactylation (Pan-Kla) levels and lactated cGAS levels in microglia were upregulated in the OGD group. Lactate reversed the effects of cGAS knockdown on microglia polarization and glycolysis. The present study reveals that the cGAS-mediated neuron injury is associated with high level of cGAS lactylation. Inhibition of cGAS promotes the M2 phenotype polarization of microglia and suppress glycolysis. Thereby, targeting cGAS provides a new strategy for the development of therapeutic agents against HIE.

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Data Availability

All data included in this study are available upon request by contact with the corresponding author.

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Acknowledgements

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Funding

This study was supported by Medical Research Project of Yancheng Municipal Health Commission (No. YK2023113).

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Author notes

  1. Lisheng Wang and Zhonghua Cai have contributed equally as co first authors.

Authors and Affiliations

  1. Department of Pediatrics, Funing County People’s Hospital, No.111, Fucheng Street, Funing County, Yancheng, 224400, China

    Lisheng Wang, Zhonghua Cai, Qi Gu & Changli Xu

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  1. Lisheng Wang
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  2. Zhonghua Cai
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  3. Qi Gu
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  4. Changli Xu
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Contributions

All authors contributed to the study conception and design. Material preparation, data collection and analysis were performed by QG and CX. The first draft of the manuscript was written by LW and ZC, LW and ZC contribute equally as co first authors. All authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.

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Correspondence to Changli Xu.

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The study was approved by the Ethics Committee of Funing County People’s Hospital. A statement to confirm that all methods were carried out in accordance with relevant guidelines and regulations.

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Supplementary Information

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10528_2023_10631_MOESM1_ESM.jpg

Supplementary file1 (JPG 368 KB)—(A) The mRNA expression and (B) protein levels of cGAS in BV2 microglia under the OGD treatment increased in a time-dependent way.

10528_2023_10631_MOESM2_ESM.jpg

Supplementary file2 (JPG 1763 KB)—Lactylation of cGAS at lysine 162 site downregulates the cGAS protein levels to promote the transformation of microglia from M1 to M2.(A) The cGAS protein levels in microglia were measured by western blot with wild type (WT) or cGAS mutants (K162R). (B–I) Secretion of IL-1β, iNOS, TNF-α, IL-4, IL-10, and Arg-1 in microglia cells of each group. ***p<0.001

Supplementary file3 (DOC 67 KB)

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Wang, L., Cai, Z., Gu, Q. et al. cGAS Deficiency Regulates the Phenotypic Polarization and Glycolysis of Microglia Through Lactylation in Hypoxic-Ischemic Encephalopathy Cell Model. Biochem Genet (2024). https://doi.org/10.1007/s10528-023-10631-2

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