胶质瘤干细胞GSLC和非干细胞nGSLC对恶性胶质瘤发生和发展分别发挥了独特的作用。我们发现EFEMP1在细胞外结合EGFR，抑制nGSLC的增殖及血管形成，却激活NOTCH通路增强GSLC的侵袭性削弱了其抗肿瘤作用。于是我们构建多个EFEMP1变体，其中一个能显著下调EGFR/AKT并阻断NOTCH介导的GSLC侵袭性。预实验结果表明据此人工合成的蛋白ZR30也具有类似的抗胶质瘤作用。因此，我们提出ZR30阻断EGFR介导的nGSLC增殖、血管发生及NOTCH介导的GSLC侵袭性，抑制胶质瘤发生发展的假设。本项目拟从体外功能实验证实ZR30对同源原代GSLC和nGSLC的抑制作用，从体内实验验证其对移植瘤发生和发展的抑制作用，阐明其阻断细胞内EGFR和NOTCH通路，降低细胞外MMP2，抑制nGSLC增殖、血管形成及阻断GSLC侵袭的作用机制，完成ZR30局部治疗胶质瘤的临床前疗效确认。

Glioma stem-like cell(GSLC) and non-glioma stem-like cell (nGSLC) play a unique role in the initiation and progression of malignant glioma respectively. We have demonstrated that EFEMP1(epidermal growth factor-containing fibulin-like extracellular matrix protein 1) suppressed proliferation and angiogenesis in nGSLC via binding to extracelluar EGFR(epidermal growth factor receptor),while enhanced the invasiveness of GSLC by activation of Notch signaling pathway, hence the tumor suppressor role of EFEMP1 is compromised. So we have constructed deletion/mutation constructs expressing various EFEMP1 variants, and one of these variants could not only downregulate expression of EGFR/AKT, but also attenuate invasiveness of GSLC via blocking NOTCH signaling pathway. Based on this variant, the synthetic protein(ZR30) have been demonstrated the similar tumor suppressor effect in the preliminary study. Therefore, we propose the hypothesis that ZR30 inhibit proliferation, invasion and angiogenesis of GSLC and nGSLC which is mediated by EGFR and NOTCH signaling pathway, resulting in the suppression of glioma initiation and progression. In this project, we will validate the tumor suppressing efficacy of ZR30 in vitro with syngeneic GSLC and nGSLC derived from glioblastoma primary culture lines, and confirm the suppressing efficacy on tumor initiation and progression in vivo with intracranial xenograft models. We will further clarify the mechanism of proliferation and angiogenesis suppression in nGSLC and invasiveness suppression in GSLC through inhibition of intracellular EGFR and NOTCH signaling pathway and attenuation of extracellular MMP2 activation. We aim to validate the preclinical therapeutic effectiveness of ZR30 with local application in glioma treatment.