剪接因子TRA2A在青蒿琥酯促进急性髓系白血病细胞凋亡中的作用及机制研究

急性髓系白血病（AML）是髓系造血干细胞恶性增殖性疾病，长期无病生存率低于30%。凋亡信号异常是AML发病及化疗耐药的一个重要科学问题。近期的研究表明靶向细胞凋亡缺陷是治疗AML新的关键策略，但目前还没有药物应用于临床。项目前期研究发现青蒿琥酯（ART）可以促进AML细胞凋亡；我们进一步通过多组学联合分析，推测ART可能是通过促进蛋白磷酸酶PP1α的表达和活化，降低剪接因子TRA2A的磷酸化，进而促进CASP8促凋亡剪接变体CASP8a的表达来诱导AML细胞凋亡。本项目将通过minigene、RIP&IP以及构建TRA2A突变体等技术方法，对ART诱导AML细胞凋亡过程中，TRA2A调节CASP8的可变剪接和PP1α调节TRA2A的磷酸化等内容进行深入研究，以解析ART促进AML细胞凋亡的新机制；并在裸鼠模型上进行验证。以期发现AML治疗的新靶点、新药物及作用机制，为其临床诊疗提供新思路。

Acute Myeloid leukemia (AML) is a malignant proliferative disease of myeloid hematopoietic stem cells. The long-term disease-free survival rate of AML is less than 30%. Aberrant apoptotic signaling is an important scientific problem in the pathogenesis and chemoresistance of AML. Recent studies have shown that targeting apoptosis defect is a new key strategy for AML treatment, but there is no drug to achieve clinical application. We previously found that artesunate can induce apoptosis in AML cells. Through additional multi-omics analysis, we speculate that ART may promote the expression and activation of protein phosphatase PP1α, which may reduce the phosphorylation level of the splicing factor TRA2A and then promote the expression of the pro-apoptotic splicing variant CASP8a of CASP8 to induce apoptosis of AML cells. In this proposal, in order to explore the new mechanism of ART induced apoptosis of AML cells, we will conduct in-depth investigation of the alternative splicing of CASP8 regulated by TRA2A and the dephosphorylation of TRA2A catalyzed by PP1α during the process of ART induced apoptosis of AML cells, by using minigene, RIP&IP, generate of TRA2A mutants and other methods. And all the results will be verified in a nude mouse model. We hope to contribute to the discovery of novel targets, drugs and mechanisms for AML therapy, and to provide new ideas for clinical diagnosis and treatment of AML.