Gαi1介导神经连接蛋白-3信号转导促胶质瘤细胞颅内生长作用及机制研究

Neuron-secretory neuroligin-3 (NLGN3) has been reported to activate multiple receptor tyrosine kinases and downstream signaling pathways required for glioma cell intracranial growth. Our studies have shown that the G protein α inhibitory subunit 1 (Gαi1) is over-expressed in human glioma tissues, and is essential in transducing signaling by several RTKs. Our preliminary studies in human glioma cells show that shRNA-mediated knockdown or CRISPR-Cas-9-mediated knockout of Gαi1 inhibited NLGN3-induced downstream Akt-mTOR and Erk activation. We propose that Gαi1 mediates NLGN3 signaling to promote glioma cell intracranial growth. In the current proposal, using multiple in vitro and in vivo biomedical assays, we will study the role and the underlying mechanisms of Gαi1 in mediating NLGN3 signaling and glioma cell growth. The potential effect of miR-200a, an anti-Gαi1 miRNA, on NLGN3-induced glioma cell growth will also be tested. We will test whether injection of lentiviral Gαi1 shRNA or lentiviral anti-Gαi1 microRNA (miR-200a), can block NLGN3-Gαi1 signaling and glioma cell intracranial growth in nude mice. These studies will demonstrate that NLGN3-Gαi1 signaling plays a key role in promoting intracranial glioma cell growth.

神经元分泌的神经连接蛋白-3（NLGN3）激活多个受体酪氨酸激酶（RTKs）和下游信号促胶质瘤细胞颅内生长。前期研究证实Gαi1（G蛋白抑制性α亚单位1）在人胶质瘤中高表达，并介导多个RTKs信号转导。预实验发现，在人胶质瘤细胞中，shRNA敲减或CRISPR-Cas9敲除Gαi1几乎阻断NLGN3诱导的下游Akt-mTOR和Erk信号活化。我们提出Gαi1介导NLGN3信号转导促胶质瘤细胞颅内生长。本项目将运用多种生物学方法，结合裸鼠胶质瘤细胞原位生长模型，解析Gαi1介导NLGN3信号转导并促胶质瘤细胞生长作用及分子机制。观察miR-200a（抗Gαi1特异性miRNA）沉默Gαi1抑制NLGN3促胶质瘤细胞生长作用。拟立体定位注射慢病毒包裹的Gαi1 shRNA或miR-200a延缓或阻断胶质瘤细胞颅内生长。拟证实NLGN3-Gαi1通路是胶质瘤细胞颅内原位生长的关键信号机制。

神经元分泌的神经连接蛋白-3（NLGN3）激活多个受体酪氨酸激酶（RTKs）和下游信号促胶质瘤细胞颅内生长。前期研究证实Gαi1/3（G蛋白抑制性α亚单位1和3）在人胶质瘤中高表达，并介导多个RTKs信号转导。实验结果证实在人胶质瘤细胞中，shRNA敲减或CRISPR-Cas9敲除Gαi1/3几乎阻断NLGN3诱导的下游Akt-mTOR和Erk信号活化。Gαi1/3介导NLGN3信号转导促胶质瘤细胞颅内生长。本项目运用了多种生物学方法并结合裸鼠胶质瘤细胞原位生长模型，解析了Gαi1/3介导NLGN3信号转导并促胶质瘤细胞生长的作用及分子机制，shRNA敲减Gαi1/3阻断了胶质瘤细胞颅内生长。证实NLGN3-Gαi通路是胶质瘤细胞颅内原位生长的关键信号机制。