The SIK1-NR4A2 axis: a core driver of epilepsy identified by integrated machine learning and experimental validation
- 1Department of Neurology, The First People’s Hospital of Qinzhou, Qinzhou, Guangxi, China
- 2Department of Neurology, The First Affiliated Hospital of Guangxi Medical University, Nanning, Guangxi, China
- 3Department of Research, The First People’s Hospital of Qinzhou, Qinzhou, Guangxi, China
Abstract
Background: Epilepsy remains a major clinical challenge, suggesting that mechanisms beyond the classical excitation-inhibition imbalance contribute to epilepsy. The Liver kinase B1 (LKB1) signaling pathway coordinates energy homeostasis and cell survival, but its key downstream effectors in epilepsy remain largely unknown.
Methods: We integrated machine learning screening of LKB1 pathway genes using public Gene Expression Omnibus datasets (GSE143272) with in vivo and in vitro experimental validation. A PTZ-induced status epilepticus rat model, AAV-mediated gene knockdown/overexpression, lentiviral transfection of primary hippocampal neurons, Electroencephalography recording, whole-cell patch-clamp recording, and Western blot analysis were employed to investigate the functional role and molecular mechanisms of candidate genes.
Results: Machine learning identified SIK1 (salt-inducible kinase 1) as the top-ranked epilepsy risk gene within the LKB1 pathway. SIK1 expression was significantly upregulated in the hippocampus of SE rats. AAV-mediated SIK1 knockdown prolonged seizure latency and reduced seizure duration, while SIK1 overexpression exacerbated seizure severity. Mechanistically, through PPI network analysis and mediation analysis, we identified NR4A2 as a downstream effector of SIK1. SIK1 negatively regulated NR4A2 expression: SIK1 knockdown increased NR4A2, whereas SIK1 overexpression decreased NR4A2. Rescue experiments showed that co-overexpression of SIK1 and NR4A2 reversed the pro-epileptic effect of SIK1 alone. Whole-cell patch-clamp recordings demonstrated that SIK1 knockdown or NR4A2 overexpression restored action potential amplitude to near-normal levels in magnesium-free-treated neurons.
Conclusion: SIK1 negatively regulates NR4A2, and restoring NR4A2 expression counteracts SIK1-induced neuronal hyperexcitability. These findings suggest that targeting the SIK1-NR4A2 axis may represent a potential therapeutic strategy for epilepsy.
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BibTeX
@article{liu2026sik1nr4a2,
title = {{The SIK1-NR4A2 axis: a core driver of epilepsy identified by integrated machine learning and experimental validation}},
author = {Xiuying Liu and Shouhuan Zheng and Kunling Yang and Lanfeng Sun and Lina Su and Shanmei Zhong and Huayue Liang and Xiaodan Luo},
journal = {Frontiers in Cell and Developmental Biology},
year = {2026},
month = oct,
doi = {10.3389/fcell.2026.1887268},
url = {https://doi.org/10.3389/fcell.2026.1887268},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - The SIK1-NR4A2 axis: a core driver of epilepsy identified by integrated machine learning and experimental validation
AU - Xiuying Liu
AU - Shouhuan Zheng
AU - Kunling Yang
AU - Lanfeng Sun
AU - Lina Su
AU - Shanmei Zhong
AU - Huayue Liang
AU - Xiaodan Luo
PY - 2026
DA - 2026/10/22
JO - Frontiers in Cell and Developmental Biology
DO - 10.3389/fcell.2026.1887268
UR - https://doi.org/10.3389/fcell.2026.1887268
AB - Background: Epilepsy remains a major clinical challenge, suggesting that mechanisms beyond the classical excitation-inhibition imbalance contribute to epilepsy. The Liver kinase B1 (LKB1) signaling pathway coordinates energy homeostasis and cell survival, but its key downstream effectors in epilepsy remain largely unknown. Methods: We integrated machine learning screening of LKB1 pathway genes using public Gene Expression Omnibus datasets (GSE143272) with in vivo and in vitro experimental validation. A PTZ-induced status epilepticus rat model, AAV-mediated gene knockdown/overexpression, lentiviral transfection of primary hippocampal neurons, Electroencephalography recording, whole-cell patch-clamp recording, and Western blot analysis were employed to investigate the functional role and molecular mechanisms of candidate genes. Results: Machine learning identified SIK1 (salt-inducible kinase 1) as the top-ranked epilepsy risk gene within the LKB1 pathway. SIK1 expression was significantly upregulated in the hippocampus of SE rats. AAV-mediated SIK1 knockdown prolonged seizure latency and reduced seizure duration, while SIK1 overexpression exacerbated seizure severity. Mechanistically, through PPI network analysis and mediation analysis, we identified NR4A2 as a downstream effector of SIK1. SIK1 negatively regulated NR4A2 expression: SIK1 knockdown increased NR4A2, whereas SIK1 overexpression decreased NR4A2. Rescue experiments showed that co-overexpression of SIK1 and NR4A2 reversed the pro-epileptic effect of SIK1 alone. Whole-cell patch-clamp recordings demonstrated that SIK1 knockdown or NR4A2 overexpression restored action potential amplitude to near-normal levels in magnesium-free-treated neurons. Conclusion: SIK1 negatively regulates NR4A2, and restoring NR4A2 expression counteracts SIK1-induced neuronal hyperexcitability. These findings suggest that targeting the SIK1-NR4A2 axis may represent a potential therapeutic strategy for epilepsy.
SN - 2296-634X
ER -