单细胞技术探讨p53与NEK2在多发性骨髓瘤中的克隆演进及耐药机制

The clonal evolution plays an very important role in replase and drug resistance in multiple myeloma (MM). p53 deletion was highly correlated with relapse and drug resistance in MM. Our previous work demonstrated p53 can directly binding NEK2 promoter and NEK2 induces drug resistance mainly through up-regulation of efflux drug pumps by activation of AKT/Wnt pathway in MM. However, the underlying mechanisms of the subclones of p53 deletion and over-expression of NEK2 involved in MM clonal evolution and molecular mechanisms are not well understood. The aims of this research are to perform the dedicated study using newly dignosed relapse and sequential patient specimens to refine the role of clonal evolution of the subclones of NEK2 and p53 during the process from Hematopoietic Stem Cell to Plasma cell lineage by single cell techniques in the development, progression, relapse and drug resistance of MM. We will perform the studies in vitro and in vivo to target p53 and/or NEK2 in MM cells, Trp53 konckout mouse and 5TGM1 MM mouse model to explore how p53 regulates NEK2 to participate drug resistance. The significance of this research will provide a profound theoretical and experimental basis to evaluate the process of clonal evolution and the pre-clinical application of the NEK2 inhibitor to overcome the drug resistance induced by p53 in MM.

克隆演进在多发性骨髓瘤（Multiple Myeloma, MM）复发及耐药中发挥着十分重要的作用。p53缺失与MM的复发和耐药密切相关。我们前期研究发现p53直接调控NEK2启动子，而NEK2通过AKT/Wnt通路激活多药耐药蛋白（MDR）参与MM的耐药，但p53缺失与NEK2高表达的亚克隆是如何参与MM的克隆演进与耐药机制尚不清楚。本项目拟收集初诊、复发及序贯临床样本，采用单细胞技术深入分析p53与NEK2亚克隆从造血干细胞到浆细胞各阶段是如何参与MM发生、发展、复发及耐药过程中的克隆演进；利用靶向p53和/或NEK2的MM细胞系、Trp53敲除小鼠和5TGM1 MM鼠模型体内及体外实验进一步探讨p53 靶向NEK2发挥其耐药的作用机制。本研究将为MM动态克隆演进学说的发展及利用NEK2抑制剂克服p53缺失介导的MM耐药患者的临床应用提供充实的理论基础和实验依据。

克隆演进在多发性骨髓瘤（Multiple Myeloma, MM）复发及耐药中发挥着十分重要的作用。p53缺失与MM的复发和耐药密切相关。我们前期研究发现p53直接调控NEK2启动子，而NEK2通过AKT/Wnt通路激活多药耐药蛋白（MDR）参与MM的耐药，但p53缺失与NEK2高表达的亚克隆是如何参与MM的克隆演进与耐药机制尚不清楚。本项目拟收集初诊、复发及序贯临床样本，采用单细胞技术深入分析p53与NEK2亚克隆从造血干细胞到浆细胞各阶段是如何参与MM发生、发展、复发及耐药过程中的克隆演进；利用靶向p53和/或NEK2的MM细胞系、Trp53敲除小鼠和5TGM1 MM鼠模型体内及体外实验进一步探讨p53 靶向NEK2发挥其耐药的作用机制。本研究将为MM动态克隆演进学说的发展及利用NEK2抑制剂克服p53缺失介导的MM耐药患者的临床应用提供充实的理论基础和实验依据。