恶性神经胶质瘤的不良疗效和预后与其组织基质蛋白的异常表达和分布关系密切。Decorin的异常与胶质瘤的浸润性和恶性度高度相关。Cyr61/CCN1是重要的细胞迁移调控分子，也是抗癌基因BATF2/Sari直接抑制的靶基因之一，从而阻滞恶性肿瘤细胞的增殖和迁移。本研究拟利用病毒载体构建的前沿技术，将特异结合Decorin的多肽表位插入腺相关病毒的衣壳蛋白，并制备靶向性的BATF2/Sari基因治疗载体，继而通过对胶质瘤细胞系和动物接种模型的感染研究，揭示其在调控细胞增殖和迁移浸润方面的疗效作用。本课题着眼于影响肿瘤细胞恶性表型的关键微环境因素，并且与靶向基因治疗的前沿技术和策略相结合，将对所制备的AAV载体在胶质瘤模型中的应用特点和潜力作出充分客观的评估。结果不仅可为新型病毒载体的研发和应用做出贡献，同时也为探求特定抗癌因子的作用机制以及深入认识胶质瘤发生发展的规律提供重要的资料和线索。

The poor therapeutic outcome and prognosis of glioma is closely related to the aberrant tissue expression and distribution of matrix proteins. Decorin, as an important cellular and matrix component, is known to be involved in the invasion of cancers, including malignant glioma. The product of BATF2/Sari anti-oncogene is able to suppress Cyr61/CCN1, which prevents the migration and metastasis of cancer cells. In this study, we proposed to use the cutting-edge technology to engineer Decorin-binding motifs into the capsid of adeno-associated virus and to achieve the targeted delivery of BATF2/Sari gene transfer in both in vitro and in vivo glioma models for the evaluation of its role in preventing cancer growth and metastasis. Meanwhile, we will explore the connections of the BATF2/Sari downstream effector Cyr61/CCN1 with the TGF-beta signal pathway, as well as the integrin related mesenchymal and matrix network. We will also conduct extensive and objective observation and estimation upon the application potential of our constructed viral vectors for the gene therapy of malignant glioma, especially in cases of higher malignancy or drug resistance. Our study focuses on the phenotype -determining key factors of the cancer microenvironment and practises the advanced technology for targeted vector construction. The results will not only produce novel therapeutical viral vectors for the treatment of malignant glioma, but also can be helpful to understand the development and progression of glioma, especially in providing useful reference and clues for the mechanism of specific cancer suppression genes.

恶性神经胶质瘤的不良疗效和预后与其组织基质蛋白的异常表达和分布关系密切。Decorin (DCN)的异常与胶质瘤的浸润性和恶性度高度相关。通过数据库分析，我们发现，DCN表达水平与胶质瘤恶性程度程正相关。但细胞学和分子生物学实验表明，DCN在胶质瘤细胞可通过诱导自噬发生，对TGF-β信号通路产生负性调控作用，从而降低胶质瘤细胞的黏附和迁移，发挥抗肿瘤活性。我们进一步探讨能否围绕DCN在胶质瘤中的高表达特点，挖掘其在肿瘤基因治疗中的应用价值。在对多种结合蛋白序列保守区的分析基础上， 筛选出具有较高活性的DCN 结合功能域DB1， 并将其融合至腺相关病毒AAV2的 衣壳蛋白VP2 编码序列的N 端； 继而利用AAV 的嵌合包装技术， 成功制备了衣壳展示DB1 表位的重组AAV。 在过表达DCN 细胞的感染实验中， 该病毒表现出针对DCN 较强的靶向性， 这为靶向高表达DCN的胶质瘤的基因治疗奠定了基础：计划利用该载体携带特定抑癌基因，展开胶质瘤基因治疗临床前研究。通过分析，我们锚定了治疗基因，即抑癌基因BATF2/Sari。我们的研究结果表明，BATF2/Sari可显著抑制肿瘤细胞增殖，促进肿瘤细胞周期发生G2/M阻滞；在胶质瘤细胞，还可通过对Wnt/β-catenin信号通路的抑制，发挥拮抗靶向治疗药物贝伐单抗诱导的胶质瘤细胞EMT和细胞迁移浸润。此外，通过ChIP-seq对其转录调控机制进一步研究发现，BATF2/Sari可通过与DNA特定模序的结合，对多种EMT相关分子的转录发挥负性调控作用；此外，BATF2/Sari还可在全基因组水平介导异染色质结构的消除，促进卫星DNA序列II的表达，这可能为其发挥肿瘤抑制活性的另一重要机制。在此基础上，我们计划利用衣壳展示DB1 表位的重组AAV携带治疗基因BATF2/Sari，将其用于恶性胶质瘤的基因治疗，相关成果可为胶质瘤的临床基因治疗提供重要的理论基础。