肿瘤微环境组成分子Netrin-4与Neuropilin-1的结合调控细胞自噬介导胶质母细胞瘤的化疗耐药及分子机制

Glioblastoma (GBM) has strong resistance to chemotherapy and recurrence, and the survival time of the patients is only 15 months. It is helpful to elucidate the mechanism of drug resistance for us to find a way to reverse drug resistance. In our previous study, we found that Netrin-4 (NTN4), as an important component of extracellular matrix, may be involved in GBM resistance to temozolomide. To study the drug resistance mechanism of GBM deeply, we learned that induced U251 resistant cell line by gene chip, has related genes up-regulated protein transport and metabolic pathways, including NTN4; tandem affinity purification and mass spectrometry analysis obtained the correlation between molecular NTN4 potential Neuropilin-1 (NRP1); and NRP1 induces autophagy in GBM; the co immunoprecipitation showed NRP1 binding with specific autophagy adapter protein p62. Based on the previous work, this study will further study NTN4/NRP1/P62, a new cascade of interacting GBM autophagy signaling pathways, and elucidate the potential mechanisms of drug resistance in clinical samples and animal models. This study will reveal the mechanism of cell matrix molecules involved in the regulation of tumor cell autophagy and chemotherapy resistance, so as to find a new way to treat the patients with recurrent GBM.

胶质母细胞瘤（GBM）具极强的化疗耐药性和复发能力，患者生存期仅15个月，阐明耐药机制有助寻找逆转耐药途径。本课题组前期研究发现，Netrin-4（NTN4）作为细胞外基质重要组分，可能参与GBM对替莫唑胺的耐药。为深入研究GBM耐药机制，我们通过基因芯片得知人工诱导的U251耐药细胞株，蛋白转运与代谢通路的相关基因上调，其中包括NTN4；串联亲和纯化和质谱分析获得了NTN4潜在关联分子Neuropilin-1（NRP1），并鉴定了二者结合促进GBM细胞自噬；初步的免疫共沉淀筛查发现NRP1与特定自噬适配蛋白p62结合。基于以上前期工作，本课题将深入研究NTN4/NRP1/P62这一新的蛋白质相互作用级联调控的GBM自噬信号通路，通过临床样本和动物模型阐明其中潜在耐药机制。该课题将揭示细胞基质分子参与调控肿瘤细胞自噬以及化疗耐药机制，以期寻找针对肿瘤微环境为靶点治疗复发胶质瘤病人的新思路。

胶质母细胞瘤（GBM）是恶性程度最高的颅内恶性肿瘤，具极强的化疗耐药性和复发能力，阐明其耐药机制有助寻找逆转GBM耐药的途径。本课题组曾经在国际上首次证明，Netrin-4（NTN4）作为细胞外基质的重要组分，可能参与GBM对替莫唑胺的耐药。为深入研究GBM耐药机制，我们通过基因芯片以及单细胞RNA测序技术证明了NTN4、NRP1、EGFR与GBM的耐药相关，进一步从细胞自噬和衰老的角度阐释NTN4诱导GBM的化疗耐药途径。串联亲和纯化和质谱分析获得了NTN4潜在关联分子Neuropilin-1（NRP1），并鉴定了NTN4/NRP1/p62这一新的蛋白质相互作用级联调控的GBM自噬信号通路；同时，描述了EGF/EGFR可以上调NTN4的表达，以及EGF/EGFR与NTN4的协同作用共抑制DNA损伤诱导的GBM细胞衰老。从自噬和衰老的角度阐述了NTN4诱导GBM细胞化疗耐药的机制，为针对肿瘤微环境为靶点治疗复发胶质瘤病人提供新思路。