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POU2F1/DNMT3a Pathway Participates in Neuropathic Pain by Hypermethylation-Mediated LRFN4 Downregulation Following Oxaliplatin Treatment

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Abstract

Evidence demonstrates that DNA methylation is associated with the occurrence and development of various neurological diseases. However, the potential target genes undergoing DNA methylation, as well as their involvement in the chemotherapy drug oxaliplatin-induced neuropathic pain, are still unclear. Here, Lrfn4, which showed hypermethylation in the promoter regions, was screened from the SRA methylation database (PRJNA587622) following oxaliplatin treatment. MeDIP and qPCR assays identified that oxaliplatin treatment increased the methylation in Lrfn4 promoter region and decreased the expression of LRFN4 in the spinal dorsal horn. The assays with gain and loss of LRFN4 function demonstrated that LRFN4 downregulation in spinal dorsal horn contributed to the oxaliplatin-induced mechanical allodynia and cold hyperalgesia. Moreover, oxaliplatin treatment increased the DNA methyltransferases DNMT3a expression and the interaction between DNMT3a and Lrfn4 promoter, while inhibition of DNMT3a prevented the downregulation of LRFN4a induced by oxaliplatin. We also observed that the transcriptional factor POU2F1 can bind to the predicted sites in DNMT3a promoter region, oxaliplatin treatment upregulated the expression of transcriptional factor POU2F1 in dorsal horn neurons. Intrathecal injection of POU2F1 siRNA prevented the DNMT3a upregulation and the LRFN4 downregulation induced by oxaliplatin. Additionally, intrathecal injection of DNMT3a siRNA or POU2F1 siRNA alleviated the mechanical allodynia induced by oxaliplatin. These findings suggested that transcription factor POU2F1 upregulated the expression of DNMT3a, which subsequently decreased LRFN4 expression through hypermethylation modification in spinal dorsal horn, thereby mediating neuropathic pain following oxaliplatin treatment.

Highlights

Downregulation of LRFN4 mediated by DNA methyltransferases DNMT3a in spinal dorsal horn contributed to the neuropathic pain induced by chemotherapy agent oxaliplatin treatment.

Transcription factor POU2F1 bound to the Dnmt3a promoter and increased the DNMT3a expression in oxliplatin-induced neuropathic pain.

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Data Availability

The data that support the findings of this study are available from the corresponding author upon reasonable request.

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Acknowledgements

This study was funded by National Natural Science Foundation of China (Grant No. 31970936), Guangdong Basic and Applied Basic Research Foundation (2023A1515030020, 2022A1515012259), Guangzhou Science and Technology Plan Project (202206060004, 202201010988).

Funding

This study was funded by National Natural Science Foundation of China (Grant No. 31970936), Guangdong Basic and Applied Basic Research Foundation (2023A1515030020, 2022A1515012259), Guangzhou Science and Technology Plan Project (202206060004, 202201010988).

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Author notes

  1. Yan-Hui Gu and Jing Wang contributed equally to this work.

Authors and Affiliations

  1. Department of General Surgery, Cancer Hospital of Zhengzhou University, 127 Dongming Rd, Zhengzhou, China

    Yan-Hui Gu, Yong Cheng, Shi-Jia Zhang, Ke-Wei Zhai & Su-Xia Luo

  2. Guangdong Province Key Laboratory of Brain Function and Disease and Zhongshan School of Medicine, Sun Yat-Sen University, Guangzhou, 510120, China

    Yan-Hui Gu, Jing Wang, Ting Xu & Wen-Jun Xin

  3. Department of Pain Management, Henan Provincial People’s Hospital, Zhengzhou University, Zhengzhou, China

    Jing Wang & Rong Tao

  4. State Key Laboratory of Oncology in Southern China, Department of Anesthesiology, Collaborative Innovation for Cancer Medicine, Sun Yat- sen University Cancer Center, Guangzhou, China

    Wei-Cheng Lu

  5. Zhongshan Medical School, Sun Yat-sen University, Zhongshan Rd. 2, Guangzhou, China

    Wen-Jun Xin

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Contributions

Su- Xia Luo and Wen-Jun Xin contributed to the study conception and design. Data collection and analysis were performed by Yan-Hui Gu and Jing Wang. The first draft of the manuscript was written by Yan-Hui Gu and Wei-Cheng Lu. Material preparation were performed Yong Cheng and Rong Tao. Experiments were performed by Yan-Hui Gu, Jing Wang, Shi-Jia Zhang, Ting Xu and Ke-Wei Zhai. All authors read and approved the final manuscript.

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Correspondence to Su-Xia Luo or Wen-Jun Xin.

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The experimental protocols were approved by the Animal Care and Use Committee of Sun Yat-sen University and conducted in strict accordance with the guideline of National Institutes of Health on the animal care and the ethical guideline.

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Gu, YH., Wang, J., Lu, WC. et al. POU2F1/DNMT3a Pathway Participates in Neuropathic Pain by Hypermethylation-Mediated LRFN4 Downregulation Following Oxaliplatin Treatment. Neurochem Res 48, 3652–3664 (2023). https://doi.org/10.1007/s11064-023-04011-w

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