CRISPR/Cas9全基因组文库筛选Venetoclax/Azacitidine耐药关键基因及其机制研究

急性髓系白血病(AML)是一种由于正常造血系统受损和非成熟细胞异常增殖与分化造成的血液疾病，AML患者的平均初诊年龄较高，经过常规化疗药物或造血干细胞移植进行治疗后可以明显改善一些AML患者的生存和预后，然而老年AML患者通常对化疗药物耐受性低且预后较差，Venetoclax和Azacitidine联合疗法近年来被用于治疗老年AML患者且疗效较好，但仍有部分患者复发或对药物不敏感。为研究和解决这个临床治疗中遇到的问题，本项目利用CRISPR/Cas9全基因组文库筛选技术，对两药联合处理下的AML细胞进行筛选，以鉴定导致耐药的关键基因，在体内外模型中验证并探索其调控机制。此外，我们还将筛选靶向目标基因及其相关通路的抑制剂，与Venetoclax和Azacitidine共同对AML细胞进行处理，以克服细胞耐药性，为改善AML复发和耐药患者的治疗方案提供新的靶点和思路。

Acute Myeloid Leukemia（AML）is a heterogeneous disease characterized by the impaired production of normal hematopoietic cells and uncontrolled proliferation and differentiation of immature blasts. The typical treatment of AML is chemotherapy with cytarabine and anthracycline drugs followed by a stem cell transplantation. However, some senior AML patients are not able to tolerate these typical chemotherapy drugs. Combinatorial therapy with Venetoclax and Azacitidine was then developed to improve the treatment efficiency for these AML patients. Despite most patients responded well to this combination therapy strategy with improved overall survival and remission rate, some patients were resistant to Venetoclax and Azacitidine or relapse after initial treatment. Here we employed a newly developed genome-wide CRISPR/Cas9 dropout screen system to identify the essential genes mediating drug resistance and relapse in vivo and in vitro with Venetoclax and Azacitidine treatment. In addition, we will investigate the mechanisms and screen for inhibitors that will form new combinatorial treatment strategies to overcome drug resistance, which will contribute to the improved treatment for AML patients with relapse or drug resistance.