温热通过YAP影响TBK1/IRF3诱导抗病毒免疫机制研究

Local hyperthermia could effectively cure human papillomavirus infected diseases.The non-targeted, distal cutaneous lesions were cured as well as the directly targeted, prompting an underlying systemic immunological antiviral responding mechanism. Our previous study found that the expression of IFN-α/β were up-regulated, while HPV E6/E7 were down-regulated, the mechanism remained to be further investigated. Recently, a view of YAP interferes TBK1/IRF3-mediated innate antiviral immunity was raised. Phosphoralation levels of YAP was up-regulated by hyperthermia in our pre-Experiment, promoting a possible mechanism that YAPTBK1/IRF3-IFN-α/β might be involved in hyperthermia induced antiviral immunity.In the following research, we will first explore the in vitro whether hyperthermia could up-regulated the phosphoralation levels of YAP to interfere the TBK1/IRF3-mediated innate antiviral immunity. Then to imitate internal environment, fresh biopsies from patients were cultured in vivo and treated by setting hyperthermia, to further verify the assumption. Furthermore, the research aims to bring further understandings into the mechanism of anti-viral immunity induced by local hyperthermia, provide a new intervene target for improving the.antiviral immunity.

温热能有效治疗HPV感染性皮肤病，治疗一处靶疣消退后，非治疗部位疣体也出现消退，这一临床现象提示温热可能诱导了系统抗病毒免疫。前期研究发现温热干预下HPV感染组织中Ⅰ型干扰素表达上调，HPV转录下调，但具体分子机制尚不十分清楚。近年来发现YAP参与了TBK1/IRF3介导的抗病毒免疫。我们在预实验中发现温热上调了YAP磷酸化水平，提示YAP- TBK1/IRF3-IFN-α/β可能参与了温热诱导的抗病毒免疫。在接下来的研究中，我们将首先通过体外实验探索温热是否通过上调YAP磷酸化水平影响TBK1/IRF3介导的抗病毒免疫及相关机制。同时通过原代组织培养模拟在体环境，进一步验证温热干预上调YAP磷酸化水平与TBK1-IRF3介导的抗病毒免疫之间的关系。本研究将有助于深入认识温热诱导抗病毒免疫机制，寻找增强机体抗病毒免疫的新的干预靶点。

前期研究发现温热干预下HPV感染组织中Ⅰ型干扰素表达上调，HPV转录下调，但具体分子机制尚不十分清楚。近年来发现YAP参与了TBK1/IRF3介导的抗病毒免疫。我们在预实验中发现温热上调了YAP磷酸化水平，提示YAP-TBK1/IRF3-IFN-α/β可能参与了温热诱导的抗病毒免疫。.研究发现：1）44℃温热干预后HPV阳性Caski细胞中IFN-α，β表达上调，YAP下游的TBK1和磷酸化TBK1，IRF3及磷酸化IRF3表达均下调，提示信号通路的失活，与干扰素表达上调表型不一致，提示温热诱导的YAP表达变化引起TBK1/IRF3活化诱导干扰素表达假说不成立。2）温热促进YAP蛋白向核内转移，一系列与细胞凋亡增殖迁移相关的下游靶基因表达发生变化，包括CTGF、Cyr61、c-Myc、BCL-2、BIRC5、EDN1/2。3）温热干预后EDN1/2表达下调，温热干预或干扰EDN1/2表达，细胞凋亡增加，温热也可以使EDN1/EDN2表达降低得到与干扰EDN1/EDN2类似的效果。温热可能通过YAP调控EDN1/EDN2表达，参与温热诱导的HPV阳性细胞的凋亡。