Service anesthésie, Gustave Roussy, Villejuif, France; Metabolomics and Cell Biology Platforms, Gustave Roussy, Villejuif, France; Equipe labellisée par la Ligue contre le cancer, Université de Paris, Sorbonne Université, INSERM UMR1138, Centre de Recherche des Cordeliers, Paris, France.
Metabolomics and Cell Biology Platforms, Gustave Roussy, Villejuif, France; Equipe labellisée par la Ligue contre le cancer, Université de Paris, Sorbonne Université, INSERM UMR1138, Centre de Recherche des Cordeliers, Paris, France; Institute for Research in Immunology and Cancer (IRIC), Université de Montréal, Montreal, Canada; Department of Medicine, Université de Montréal, Montreal, Quebec, Canada.
Methods Cell Biol. 2022;172:83-98. doi: 10.1016/bs.mcb.2022.01.003. Epub 2022 Mar 12.
Immunogenic cell death (ICD) is a modality of cellular demise that when it is induced by certain anticancer treatments can ignite an adaptive anticancer immune response. ICD is characterized by the emission of a specific set of danger-associated molecular patterns (DAMPs) including calreticulin exposure at the plasma membrane, ATP liberation, HMGB1 exodus and type-I IFN release. The apical signaling triggering the appearance of these hallmarks involves the phosphorylation on serine 51 of the α-subunit of eukaryotic initiation factor 2 (EIF2), a key protein in the orchestration of endoplasmic reticulum (ER) stress responses. EIF2α can be phosphorylated by a family of four EIF2A kinases: EIF2AK1-4 (best known as heme regulated inhibitor, HRI, protein kinase R, PKR, protein kinase R-like endoplasmic reticulum kinase, PERK, and general control non-derepressible 2, GCN2), that each respond to a specific type of cellular stress. Here, we describe different techniques to investigate the biochemical pathways leading to eIF2α phosphorylation in the context of ICD.
免疫原性细胞死亡（ICD）是一种细胞死亡方式，当它被某些抗癌治疗所诱导时，可以引发适应性抗癌免疫反应。ICD 的特征是释放一组特定的危险相关分子模式（DAMPs），包括质膜上钙网蛋白的暴露、ATP 的释放、HMGB1 的外排以及 I 型 IFN 的释放。触发这些特征出现的顶端信号涉及真核起始因子 2（EIF2）α亚基丝氨酸 51 的磷酸化，EIF2 是内质网（ER）应激反应协调的关键蛋白。EIF2α 可以被四种 EIF2A 激酶之一磷酸化：EIF2AK1-4（最著名的是血红素调节抑制剂（HRI）、蛋白激酶 R（PKR）、蛋白激酶 R 样内质网激酶（PERK）和一般控制非阻遏物 2（GCN2）），它们分别对特定类型的细胞应激作出反应。在这里，我们描述了不同的技术来研究 ICD 背景下导致 eIF2α 磷酸化的生化途径。
