Liu Bin, Zhu Lin, Zhang Caijuan, Wang Dunfang, Liu Haifan, Liu Jianyao, Sun Jingwei, Feng Xue, Yang Weipeng
State Key Laboratory for Quality Ensurance and Sustainable Use of Dao-di Herbs, Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100010, China.
School of Traditional Chinese Medicine, Beijing University of Chinese Medicine, Beijing 100029, China.
Int J Mol Sci. 2025 Sep 2;26(17):8533. doi: 10.3390/ijms26178533.
Influenza A virus (IAV) infection triggers excessive activation of PANoptosis-a coordinated form of programmed cell death integrating pyroptosis, apoptosis, and necroptosis-which contributes to severe immunopathology and acute lung injury. However, the molecular regulators that drive PANoptosis during IAV infection remain poorly understood. In this study, we integrated bulk and single-cell RNA sequencing (scRNA-seq) datasets to dissect the cellular heterogeneity and transcriptional dynamics of PANoptosis in the influenza-infected lung. PANoptosis-related gene activity was quantified using the AUCell, ssGSEA, and AddModuleScore algorithms. Machine learning approaches, including Support Vector Machine (SVM), Random Forest (RF), and Least Absolute Shrinkage and Selection Operator (LASSO) regression, were employed to identify key regulatory genes. scRNA-seq analysis revealed that PANoptosis activity was primarily enriched in macrophages and neutrophils. Integration of transcriptomic and computational data identified cathepsin B (CTSB) as a central regulator of PANoptosis. In vivo validation in an IAV-infected mouse model confirmed elevated expression of PANoptosis markers and upregulation of CTSB. Mechanistically, CTSB may facilitate NLRP3 inflammasome activation and promote lysosomal dysfunction-associated inflammatory cell death. These findings identify CTSB as a critical mediatoCTSBr linking lysosomal integrity to innate immune-driven lung injury and suggest that targeting CTSB could represent a promising therapeutic strategy to alleviate influenza-associated immunopathology.
甲型流感病毒（IAV）感染会引发PANoptosis的过度激活，PANoptosis是一种程序性细胞死亡的协调形式，整合了焦亡、凋亡和坏死性凋亡，这会导致严重的免疫病理和急性肺损伤。然而，在IAV感染期间驱动PANoptosis的分子调节因子仍知之甚少。在本研究中，我们整合了大量和单细胞RNA测序（scRNA-seq）数据集，以剖析流感感染肺部中PANoptosis的细胞异质性和转录动态。使用AUCell、ssGSEA和AddModuleScore算法对PANoptosis相关基因活性进行量化。采用机器学习方法，包括支持向量机（SVM）、随机森林（RF）和最小绝对收缩和选择算子（LASSO）回归，来识别关键调控基因。scRNA-seq分析表明，PANoptosis活性主要富集于巨噬细胞和中性粒细胞中。转录组学和计算数据的整合确定组织蛋白酶B（CTSB）为PANoptosis的核心调节因子。在IAV感染的小鼠模型中进行的体内验证证实了PANoptosis标志物的表达升高和CTSB的上调。从机制上讲，CTSB可能促进NLRP3炎性小体的激活，并促进与溶酶体功能障碍相关的炎性细胞死亡。这些发现确定CTSB是连接溶酶体完整性与先天免疫驱动的肺损伤的关键介质，并表明靶向CTSB可能是一种有前景的治疗策略，以减轻流感相关的免疫病理。
