PGK1 inhibitor CBR-470-1 protects neuronal cells from MPP+
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Jinyu Zheng1, Jian-Liang Zhu2, Yufeng Zhang3
1Department of Neurosurgery, The Affiliated Huai'an Hospital of Xuzhou Medical University, Huai'an, China.
Abstract:
The neurotoxin MPP+ (1-methyl-4-phenylpyridinium ion) disrupts mitochondrial function leading to oxidative stress and neuronal death. Here we examine whether activation of the Keap1-Nrf2 cascade can protect SH-SY5Y neuroblastoma cells from MPP+-induced cytotoxicity. Treatment of SH-SY5Y cells with CBR-470-1, an inhibitor of the glycolytic enzyme phosphoglycerate kinase 1 (PGK1), leads to methylglyoxal modification of Keap1, Keap1-Nrf2 disassociation, and increased expression of Nrf2 responsive genes. Pretreatment with CBR-470-1 potently attenuated MPP+-induced oxidative injury and SH-SY5Y cell apoptosis. CBR-470-1 neuroprotection is dependent upon Nrf2, as Nrf2 shRNA or CRISPR/Cas9-mediated Nrf2 knockout, abolished CBR-470-1-induced SH-SY5Y cytoprotection against MPP+. Consistent with these findings, PGK1 depletion or knockout mimicked CBR-470-1-induced actions and rendered SH-SY5Y cells resistant to MPP+-induced cytotoxicity. Furthermore, activation of the Nrf2 cascade by CRISPR/Cas9-induced Keap1 knockout protected SH-SY5Y cells from MPP+. In Keap1 or PGK1 knockout SH-SY5Y cells,CBR-470-1 failed to offer further cytoprotection against MPP+. Collectively PGK1 inhibition by CBR-470-1 protects SH-SY5Y cells from MPP+ via activation of the Keap1-Nrf2 cascade.
Insights
Inhibiting phosphoglycerate kinase 1 (PGK1) with CBR-470-1 protects neuroblastoma cells from MPP+-induced damage. This neuroprotection occurs through activating the Keap1-Nrf2 pathway, crucial for cellular defense against oxidative stress.
Area of Science:
- Neuroscience
- Cell Biology
- Biochemistry
Background:
- The neurotoxin MPP+ induces mitochondrial dysfunction, oxidative stress, and neuronal death, contributing to neurodegenerative conditions.
- The Keap1-Nrf2 signaling pathway is a key regulator of cellular antioxidant responses.
- SH-SY5Y neuroblastoma cells are a common model for studying neuronal toxicity and potential therapeutic interventions.
Purpose of the Study:
- To investigate whether activating the Keap1-Nrf2 cascade can protect SH-SY5Y neuroblastoma cells against MPP+-induced cytotoxicity.
- To determine the role of phosphoglycerate kinase 1 (PGK1) inhibition in modulating the Keap1-Nrf2 pathway and conferring neuroprotection.
Main Methods:
- SH-SY5Y cells were treated with CBR-470-1, a PGK1 inhibitor, followed by exposure to MPP+.
- Nrf2 and Keap1 gene expression and protein interactions were analyzed.
- Gene knockdown (shRNA) and knockout (CRISPR/Cas9) strategies were employed to assess the necessity of Nrf2 and Keap1.
- Cell viability assays and apoptosis measurements were conducted to evaluate cytotoxicity.
Main Results:
- CBR-470-1 treatment induced methylglyoxal modification of Keap1, leading to Keap1-Nrf2 disassociation and increased Nrf2 target gene expression.
- Pretreatment with CBR-470-1 significantly reduced MPP+-induced oxidative injury and apoptosis in SH-SY5Y cells.
- Neuroprotection by CBR-470-1 was dependent on Nrf2, as Nrf2 depletion or knockout abolished the protective effect.
- PGK1 depletion or knockout mimicked CBR-470-1's protective effects, and Keap1 knockout also conferred resistance to MPP+.
- Further treatment with CBR-470-1 did not enhance protection in Keap1 or PGK1 knockout cells.
Conclusions:
- Inhibition of PGK1 by CBR-470-1 effectively protects SH-SY5Y neuroblastoma cells from MPP+-induced neurotoxicity.
- This neuroprotection is mediated through the activation of the Keap1-Nrf2 signaling cascade.
- Targeting PGK1 represents a potential therapeutic strategy for neurodegenerative diseases involving oxidative stress.
