温热通过DnaJA4和CRAC调控NLRP3/IL-1β治疗病毒疣机制研究

Hyperthermia is a noninvasive method which has been successfully used in the treatment of warts with a low recurrence rate. However, the mechanism of hyperthermia remains unclear, and some patients have no response to hyperthermia. Our previous study showed that local inflammation occurred during the treatment of hyperthermia, and the inflammatory cytokine IL-1β expression level was correlated to the patients’ response to hyperthermia. After the priming and activation, NLRP3 inflammasome can secrete inflammatory cytokines including IL-1β, and induce pyroptosis. Preliminary results showed that DnaJA4, a member of heat shock proteins, may inhibit the priming of NLRP3 inflammasome and affect the expression of IL-1β during hyperthermia. Besides, hyperthermia may activate NLRP3 inflammasome by calcium-release-activated calcium channel (CRAC). This project will focus on the mechanism of hyperthermia regulating the priming and activation of NLRP3 inflammasome. The effect of DnaJA4 and CRAC which regulats hyperthermia induced NLRP3 priming and activation will be further investigated by cellular and animal studies. Local inflammation may be exacerbated by the interfere of DnaJA4 and CRAC, which may improve the hyperthermia therapeutic efficacy as expected.

温热治疗病毒疣有良好的临床疗效和无创、低复发等优势，但作用机制尚不明确，且部分患者无反应。课题组前期发现温热能诱导局部炎症，且温热诱导炎症因子IL-1β分泌水平与患者对温热治疗的反应性相关。NLRP3炎症小体启动激活后可分泌IL-1β等炎症因子并诱导细胞焦亡。预实验结果提示温热治疗中热休克蛋白DnaJA4可能阻碍了NLRP3启动进而影响IL-1β表达，温热还可能通过钙释放激活钙通道(CRAC)促进NLRP3激活。本项目将以温热对角质形成细胞NLRP3炎症小体启动激活的调控作用为研究核心，在细胞、组织和动物水平研究DnaJA4和CRAC在温热诱导NLRP3启动和激活中的作用机制；并期望通过干预DnaJA4或CRAC促进温热诱导的局部炎症，进而提升治疗效果。

温热治疗病毒疣有良好的临床疗效和无创、低复发等优势，但作用机制尚不明确，且部分患者无反应。课题组前期发现温热能诱导局部炎症，且温热诱导炎症因子IL-1β分泌水平与患者对温热治疗的反应性相关。本研究目前发现44℃温热诱导角质形成细胞内NLRP3/IL-1β炎症小体通路的启动，诱导NLRP3启动通路中主要的几个基因包括NLRP3，ASC, Caspase 1, IL-1β等mRNA的转录增加；发现44℃温热诱导角质形成细胞通过影响Ca2+离子内流，氧化应激，促进NLRP3炎症小体通路的激活，诱导NLRP3启动通路中主要的几个基因包括NLRP3，Caspase 1, IL-1β等的蛋白表达增加，并促进细胞死亡；发现敲除DnaJA4能够显著促进温热诱导的NLRP3炎症小体的启动激活和角质形成细胞死亡；发现44℃温热影响包括TRPM2在内的几种温度敏感性离子通道mRNA的转录；通过应用TRPM2通道激活剂以及抑制剂，能够分别促进和抑制温热诱导的NLRP3炎症小体激活。未来期望通过调控TRPM2等温度敏感性离子通道促进局部炎症，进而提升治疗效果。