海绵来源的新型肿瘤细胞分化诱导分子的发现及其作用机制

诱导肿瘤细胞分化是白血病治疗公认的有效策略。本项目针对致死率高、治疗药物匮乏的急性髓系白血病(AML)，通过构建基于分化转录启动子(MCSFR)的可表达绿色荧光蛋白GFP的AML细胞分化筛选模型(改造的细胞可荧光显色活性成分)，高效筛选具有诱导分化效应的活性分子。前期从3种海绵中已追踪得到3个新型生物碱可促使AML细胞上调分化抗原CD11c和CD11b，但其诱导分化的作用机制尚不明确。本项目拟结合UPLC-DAD-MS/MS分子指纹图谱－特征谱学分析等多维识别手段，进一步从中发掘具有诱导肿瘤细胞分化活性的新分子，综合运用Biotin偶联，Pull-down assay结合质谱分析，SPR，ITC等化学生物学方法探寻其作用靶点，并采用高内涵细胞分析，RNA干扰等方法系统阐明其诱导细胞分化的机制，融合多学科方法技术协同创新，为开发基于诱导肿瘤细胞分化治疗的新型海洋药物夯实基础。

Inducing tumor cell differentiation is a recognized effective strategy for leukemia treatment. This project is aimed at acute myeloid leukemia (AML), which is high lethality and lack of therapeutic drugs. The applicant has constructed an AML cell differentiation screening model based on the macrophage colony-stimulating factor receptor (MCSFR) that can express the green fluorescent protein GFP, and the modified cells can be used for screening chemical components with inducing differentiation activity by fluorescent coloration. The applicant discovered 3 alkaloids from 3 sponge extracts had the bioactivity of inducing AML cell differentiation, which can promote tumor cells to up-regulate the differentiation antigens CD11c and CD11b, but the mechanism of action is unknown. The applicant intends to further discover new bioactive molecules with the bioactivity of inducing tumor cell differentiation by using UPLC-DAD-MS/MS molecular fingerprinting-characteristic analysis and multi-dimensional identification methods. Comprehensively using chemical biological methods such as Biotin coupling, Pull-down assay combined with mass spectrometry, SPR, ITC etc to explore bioactive compound’s action target. Utilizing high-content cell analysis, RNA interference and other methods to systematically elucidate its action mechanism of inducing AML cell differentiation. The integration of multi-disciplinary methods and technologies for collaborative innovation will lay a solid foundation for R&D of new marine drugs based on inducing tumor cell differentiation.