溶瘤新城疫病毒引起肺癌细胞免疫性细胞死亡的分子机制研究

Oncolytic Newcastle disease virus (NDV) induces immunogenic cell death (ICD) in infected cancer cells and elicits anticancer immune response. We recently reported that NDV induces drug-resistant lung cancer cell death, releasing HMGB1 (BMC cancer, 2014). Our preliminary data showed that NDV activates STAT1 and MAPK pathways in lung cancer cells, and releases HMGB1 and HSP90, suggesting that NDV may cause ICD. However, the mechanism remains unclear. Therefore, to uncover the underlying mechanism by which NDV induces ICD in lung cancer cells, we will examine the morphological changes and biochemical indexes of ICD such as calreticulin surface exposure in NDV-infected lung cancer cells. In addition, the difference in gene expression and alteration of canonical signal pathways will be examined between infected and uninfected lung cancer cells and the signaling pathways responsible for NDV-induced ICD will be elucidated. Furthermore, by knocking down the specific key genes regulating NDV-induced ICD, their roles in ICD will be dissected. Moreover, by determing the immune factors releasing from in NDV-infected lung cancer cells as well as the quantity of immune cells in blood from nude mice bearing subcutaneous tumors, the effect of NDV-induced ICD in anti-tumor immunity will be explored. Together, our study will provide a novel strategy for NDV-mediated biological treatment in lung cancer.

溶瘤新城疫病毒（NDV）可引起肿瘤细胞免疫性细胞死亡（immunogenic cell death，ICD），激发机体的抗肿瘤免疫应答。申请人近期研究表明，NDV诱导耐药肺癌细胞死亡，释放HMGB1（BMC cancer, 2014）。预实验结果显示，NDV感染肺癌细胞活化STAT1和MAPK通路，释放HMGB1和HSP90，提示NDV可能引起ICD，但其作用机制不详。申请人拟检测NDV感染肺癌细胞引起ICD的形态学变化和生化指标，包括钙网蛋白外翻等，并检测感染和未感染NDV的肺癌细胞基因表达和信号通路活化差异，揭示 NDV引起肺癌细胞ICD的信号通路。通过干扰上游基因验证其调控靶点，进一步阐明其分子机制。此外，通过检测感染NDV的肺癌细胞释放的免疫因子，以及注射NDV的皮下瘤裸鼠血液中免疫细胞数量，探讨NDV引起的ICD在抗肿瘤免疫中的作用。本研究将为NDV用于肺癌生物治疗提供新的策略。

溶瘤新城疫病毒（Newcastle disease virus，NDV），一种禽类副粘病毒，可在多种肿瘤中复制，并具有强的细胞毒性杀伤力。除了直接溶瘤外，溶瘤新城疫病毒还触发免疫原性细胞死亡（ICD）并引发抗肿瘤免疫。我们研究表明：1. 溶瘤NDV诱导肺癌细胞自噬和凋亡。2. NDV诱导肺癌细胞钙网蛋白外翻。3. NDV诱导肺癌细胞HMGB1和HSP70/90的释放。4. 敲低自噬关键基因，减弱NDV诱导的免疫性细胞死亡。5. NDV感染细胞产生的上清可抑制裸鼠皮下瘤生长。总而言之，本课题表明溶瘤性NDV是一种有效的免疫性细胞死亡诱导剂，并且自噬促进NDV介导的肺癌细胞免疫性细胞死亡。本研究通过阐明NDV引起肺癌细胞免疫性细胞死亡的分子机制，为NDV制定合理的临床肿瘤生物治疗方案提供理论依据和新思路。