Oś SIK1-NR4A2: główny czynnik napędzający epilepsję zidentyfikowany za pomocą uczenia maszynowego i potwierdzenia eksperymentalnego
The SIK1-NR4A2 axis: a core driver of epilepsy identified by integrated machine learning and experimental validation
W skrócie
Naukowcy za pomocą sztucznej inteligencji i badań na zwierzętach odkryli, że białko SIK1 odgrywa kluczową rolę w powstawaniu napadów epilepsji poprzez kontrolę innego białka o nazwie NR4A2. Gdy zredukowano ilość SIK1 w mózgu, napady były słabsze i rzadsze, a gdy go zwiększono, napady były bardziej groźne. Odkrycie to sugeruje, że leczenie ukierunkowane na zmianę poziomu tych dwóch białek mogłoby pomóc pacjentom z epilepsją.
Oryginalny abstract (angielski)
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 and 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.