胶质母细胞瘤（GBM）作为恶性程度最高,也是最常见的原发性中枢神经系统肿瘤，其治疗与预后多年没有大的改善。新兴的程序性死亡受体1（PD-1）抑制免疫疗法可能是GBM治疗困境的破局者，然而其效能可能抑制了其效果的充分展现。将T淋巴细胞中的PD-1的编码基因PDCD1利用CRISPR/Cas9技术敲除能达到与PD-1抑制类似的T淋巴细胞激活效果,且其作用更加持久。申请人前期研究发现利用聚乙二醇/聚赖氨酸聚合物进行基因编辑效率高，安全性好，而本项目预实验利用特异靶向CD3受体的聚乙二醇/聚赖氨酸聚合物PDCD1 CRISPR/Cas9系统敲除T淋巴细胞PD-1表达，激活了T淋巴细胞并提高了其杀伤GBM细胞的效果，且本系统稳定性高，安全性好。本研究拟在此基础上研究本聚合物CRISPR/Cas9系统的体内GBM杀伤效果，并初步研究其机制。这将为基于T淋巴细胞的GBM免疫治疗提供新的手段和思路。

As the commonest and most malignant primary central nervous system tumor, glioblastoma has defied the improvement of its management and prognosis for decades. Recently, immunotherapy pertaining to inhibition of programmed death 1 (PD-1) has emerged as a promising approach to resolve the current stalemate in glioblastoma (GBM) treatment, nonetheless, the current PD-1 based therapeutic tools in a dearth of permanent PD-1 knock-off were believed to not fulfill the tempting potential of PD-1-based therapy in GBM management. Strategically knocking out of PDCD1 genome (the programming gene for PD-1) using CRISPR/Cas 9 is postulated to afford a similar consequence as does PD-1 inhibition, which may also produce longer activation of T lymphocytes. Previously, the applicant’s works validated the utilities of polyethylene glycol (PEG)/polylysine (pLys) for construction of an efficient and safe delivery system for therapeutic nucleic acids. Our preliminary experiment utilized a CD3-targeted PEG/pLys polymer-CRISPR/Cas 9 system, which could result appreciable targeting activity specifically to T-lymphocytes, capable of silencing the expression of PD-1 in these T-lymphocytes, followed by T-lymphocytes activation. In consequence, the activated T-lymphocytes effectively induced the death of GBM cells. In reference to the aforementioned results, the current study aims to explore the insight into its in vivo performance, and the underlying molecular mechanism. To sum up, a synthetic ensemble of genetic, nanotechnology and immunotherapy is projected to press the arch of glioblastoma management paradigm, emphasizing T-lymphocyte-based GBM immunotherapy as a novel therapeutic avenue.