Regulated cell death is a fate of cells in (patho)physiological conditions during which extrinsic or intrinsic signals or redox equilibrium pathways following infection, cellular stress or injury are coupled to cell death modalities like apoptosis, necroptosis, pyroptosis or ferroptosis. An immediate survival response to cellular stress is often induction of autophagy, a process that deals with removal of aggregated proteins and damaged organelles by a lysosomal recycling process. These cellular processes and their regulation are crucial in several human diseases. Exploiting high-throughput assays which discriminate distinct cell death modalities and autophagy are critical to identify potential therapeutic agents that modulate these cellular responses. In the past few years, luciferase-based assays have been widely developed for assessing regulated cell death and autophagy pathways due to their simplicity, sensitivity, known chemistry, different spectral properties and high-throughput potential. Here, we review basic principles of bioluminescent reactions from a mechanistic perspective, along with their implication in vitro and in vivo for probing cell death and autophagy pathways. These include applying luciferase-, luciferin-, and ATP-based biosensors for investigating regulated cell death modalities. We discuss multiplex bioluminescence platforms which simultaneously distinguish between the various cell death phenomena and cellular stress recovery processes such as autophagy. We also highlight the recent technological achievements of bioluminescent tools for the prediction of drug effectiveness in pathways associated with regulated cell death.

中文翻译：

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在受调节的细胞死亡方式和自噬中利用荧光素酶化学：概述和前景


调节性细胞死亡是细胞在（病理）生理条件下的命运，在此期间，感染、细胞应激或损伤后的外源性或内源性信号或氧化还原平衡途径与细胞死亡方式（如细胞凋亡、坏死性凋亡、焦亡或铁死亡）耦合。对细胞应激的即时生存反应通常是诱导自噬，自噬是一个通过溶酶体回收过程去除聚集蛋白质和受损细胞器的过程。这些细胞过程及其调节在几种人类疾病中至关重要。利用区分不同细胞死亡方式和自噬的高通量检测对于确定调节这些细胞反应的潜在治疗剂至关重要。在过去的几年中，基于荧光素酶的检测方法因其简单性、灵敏度、已知的化学性质、不同的光谱特性和高通量潜力而被广泛开发用于评估受调节的细胞死亡和自噬途径。在这里，我们从机理的角度回顾了生物发光反应的基本原理，以及它们在体外和体内探测细胞死亡和自噬途径的影响。这些应用包括应用基于荧光素酶、荧光素和 ATP 的生物传感器来研究受调节的细胞死亡模式。我们讨论了多重生物发光平台，该平台同时区分各种细胞死亡现象和细胞应激恢复过程，例如自噬。我们还强调了生物发光工具的最新技术成就，用于预测与调节细胞死亡相关的途径中的药物有效性。