Department of Respiratory Medicine, Shenzhen Longhua District Central Hospital, Affiliated Central Hospital of Shenzhen Longhua District, Guangdong Medical University, Shenzhen, 518110, China.
The State Key Lab of Respiratory Disease, The First Affiliated Hospital, The Institute for Chemical Carcinogenesis, School of Public Health, Guangzhou Medical University, Guangzhou, 510182, China.
Cell Death Dis. 2022 Feb 25;13(2):188. doi: 10.1038/s41419-022-04633-y.
Receptor-interacting protein kinase 3 (RIPK3) functions as a central regulator of necroptosis, mediating signaling transduction to activate pseudokinase mixed lineage kinase domain-like protein (MLKL) phosphorylation. Increasing evidences show that RIPK3 contributes to the pathologies of inflammatory diseases including multiple sclerosis, infection and colitis. Here, we identified a novel small molecular compound Salt-inducible Kinases (SIKs) inhibitor HG-9-91-01 inhibiting necroptosis by targeting RIPK3 kinase activity. We found that SIKs inhibitor HG-9-91-01 could block TNF- or Toll-like receptors (TLRs)-mediated necroptosis independent of SIKs. We revealed that HG-9-91-01 dramatically decreased cellular activation of RIPK3 and MLKL. Meanwhile, HG-9-91-01 inhibited the association of RIPK3 with MLKL and oligomerization of downstream MLKL. Interestingly, we found that HG-9-91-01 also trigger RIPK3-RIPK1-caspase 1-caspase 8-dependent apoptosis, which activated cleavage of GSDME leading to its dependent pyroptosis. Mechanistic studies revealed that SIKs inhibitor HG-9-91-01 directly inhibited RIPK3 kinase activity to block necroptosis and interacted with RIPK3 and recruited RIPK1 to activate caspases leading to cleave GSDME. Importantly, mice pretreated with HG-9-91-01 showed resistance to TNF-induced systemic inflammatory response syndrome. Consistently, HG-9-91-01 treatment protected mice against Staphylococcus aureus-mediated lung damage through targeting RIPK3 kinase activity. Overall, our results revealed that SIKs inhibitor HG-9-91-01 is a novel inhibitor of RIPK3 kinase and a potential therapeutic target for the treatment of necroptosis-mediated inflammatory diseases.
受体相互作用蛋白激酶 3（RIPK3）作为细胞坏死的核心调节因子，通过介导信号转导来激活假激酶混合谱系激酶结构域样蛋白（MLKL）磷酸化。越来越多的证据表明，RIPK3 参与了包括多发性硬化症、感染和结肠炎在内的炎症性疾病的病理过程。在这里，我们发现了一种新型小分子化合物 Salt-inducible Kinases（SIKs）抑制剂 HG-9-91-01，它可以通过靶向 RIPK3 激酶活性来抑制细胞坏死。我们发现 SIKs 抑制剂 HG-9-91-01 可以阻断 TNF 或 Toll 样受体（TLRs）介导的细胞坏死，而不依赖于 SIKs。我们揭示了 HG-9-91-01 可以显著降低 RIPK3 和 MLKL 的细胞活性。同时，HG-9-91-01 抑制了 RIPK3 与 MLKL 的结合以及下游 MLKL 的寡聚化。有趣的是，我们发现 HG-9-91-01 还触发了 RIPK3-RIPK1-半胱天冬酶 1-半胱天冬酶 8 依赖性细胞凋亡，该凋亡激活了 GSDME 的切割，从而导致其依赖性细胞焦亡。机制研究表明，SIKs 抑制剂 HG-9-91-01 通过直接抑制 RIPK3 激酶活性来阻断细胞坏死，并与 RIPK3 相互作用并招募 RIPK1 来激活半胱天冬酶，导致 GSDME 的切割。重要的是，用 HG-9-91-01 预处理的小鼠对 TNF 诱导的全身炎症反应综合征表现出抗性。一致地，HG-9-91-01 通过靶向 RIPK3 激酶活性来保护小鼠免受金黄色葡萄球菌介导的肺部损伤。总的来说，我们的研究结果揭示了 SIKs 抑制剂 HG-9-91-01 是 RIPK3 激酶的一种新型抑制剂，也是治疗细胞坏死介导的炎症性疾病的潜在治疗靶点。
