Department of Immunobiology, College of Life Science and Technology, Jinan University, Guangzhou, 510632, China.
Department of Fetal Medicine, the First Affiliated Hospital of Jinan University, Guangzhou, 510630, China.
Acta Pharmacol Sin. 2024 Mar;45(3):594-608. doi: 10.1038/s41401-023-01182-8. Epub 2023 Nov 14.
PANoptosis is a new type of cell death featured with pyroptosis, apoptosis and necroptosis, and is implicated in organ injury and mortality in various inflammatory diseases, such as sepsis and hemophagocytic lymphohistiocytosis (HLH). Reverse electron transport (RET)-mediated mitochondrial reactive oxygen species (mtROS) has been shown to contribute to pyroptosis and necroptosis. In this study we investigated the roles of mtROS and RET in PANoptosis induced by TGF-β-activated kinase 1 (TAK1) inhibitor 5Z-7-oxozeaenol (Oxo) plus lipopolysaccharide (LPS) as well as the effects of anti-RET reagents on PANoptosis. We showed that pretreatment with anti-RET reagents 1-methoxy PMS (MPMS) or dimethyl fumarate (DMF) dose-dependently inhibited PANoptosis in macrophages BMDMs and J774A.1 cells induced by Oxo/LPS treatment assayed by propidium iodide (PI) staining. The three arms of the PANoptosis signaling pathway, namely pyroptosis, apoptosis and necroptosis signaling, as well as the formation of PANoptosomes were all inhibited by MPMS or DMF. We demonstrated that Oxo/LPS treatment induced RET and mtROS in BMDMs, which were reversed by MPMS or DMF pretreatment. Interestingly, the PANoptosome was co-located with mitochondria, in which the mitochondrial DNA was oxidized. MPMS and DMF fully blocked the mtROS production and the formation of PANoptosome induced by Oxo plus LPS treatment. An HLH mouse model was established by poly(I:C)/LPS challenge. Pretreatment with DMF (50 mg·kg·d, i.g. for 3 days) or MPMS (10 mg·kg·d, i.p. for 2 days) (DMF i.g. MPMS i.p.) effectively alleviated HLH lesions accompanied by decreased hallmarks of PANoptosis in the liver and kidney. Collectively, RET and mtDNA play crucial roles in PANoptosis induction and anti-RET reagents represent a novel class of PANoptosis inhibitors by blocking oxidation of mtDNA, highlighting their potential application in treating PANoptosis-related inflammatory diseases. PANoptotic stimulation induces reverse electron transport (RET) and reactive oxygen species (ROS) in mitochondia, while 1-methoxy PMS and dimethyl fumarate can inhibit PANoptosis by suppressing RETmediated oxidation of mitochondrial DNA.
细胞发生潘氏凋亡时会同时发生细胞焦亡、细胞凋亡和细胞坏死，这与各种炎症性疾病（如脓毒症和噬血细胞性淋巴组织细胞增多症）中的器官损伤和死亡率有关。已经表明，逆向电子传递（RET）介导的线粒体活性氧（mtROS）有助于细胞焦亡和细胞坏死。在这项研究中，我们研究了 TGF-β激活激酶 1（TAK1）抑制剂 5Z-7-氧杂氮卓醇（Oxo）加脂多糖（LPS）诱导的潘氏凋亡中 mtROS 和 RET 的作用，以及抗 RET 试剂对潘氏凋亡的影响。我们表明，用抗 RET 试剂 1-甲氧基 PMS（MPMS）或二甲基富马酸（DMF）预处理可剂量依赖性地抑制 Oxo/LPS 处理诱导的巨噬细胞 BMDMs 和 J774A.1 细胞的潘氏凋亡，通过碘化丙啶（PI）染色进行测定。潘氏凋亡信号通路的三个分支，即细胞焦亡、细胞凋亡和细胞坏死信号，以及潘氏凋亡体的形成，均被 MPMS 或 DMF 抑制。我们证明，Oxo/LPS 处理诱导 BMDMs 中的 RET 和 mtROS，而 MPMS 或 DMF 预处理可逆转这种作用。有趣的是，潘氏凋亡体与线粒体共定位，其中线粒体 DNA 被氧化。MPMS 和 DMF 完全阻断了 Oxo 加 LPS 处理诱导的 mtROS 产生和潘氏凋亡体的形成。通过 poly(I:C)/LPS 挑战建立了 HLH 小鼠模型。用 DMF（50 mg·kg·d，灌胃 3 天）或 MPMS（10 mg·kg·d，腹腔注射 2 天）预处理（DMF 灌胃，MPMS 腹腔注射）可有效缓解 HLH 病变，同时减少肝脏和肾脏中潘氏凋亡的特征。总之，RET 和 mtDNA 在潘氏凋亡诱导中起着关键作用，抗 RET 试剂通过阻断 mtDNA 的氧化，代表了一类新型的潘氏凋亡抑制剂，为治疗潘氏凋亡相关炎症性疾病提供了潜在的应用。潘氏凋亡刺激诱导线粒体中的逆向电子传递（RET）和活性氧（ROS），而 1-甲氧基 PMS 和二甲基富马酸可以通过抑制 RET 介导的线粒体 DNA 氧化来抑制潘氏凋亡。
