NR3C1通过调控糖酵解参与B淋巴细胞发育的作用及机制研究

申请人课题组前期研究及大量临床证据表明糖皮质激素受体基因（NR3C1）缺失性突变是导致急性淋巴细胞白血病（ALL）难治和复发的重要因素之一。但NR3C1对正常造血分化的作用及调控机制尚不清晰。申请人前期通过构建造血干细胞条件性Nr3c1敲除小鼠，发现敲除小鼠B淋巴细胞发育异常，B淋巴前体细胞和共同淋系祖细胞显著性增加，该表型与人类急性B淋巴细胞白血病发生过程中B淋系祖细胞的明显扩增相吻合。我们进一步的预实验提示Nr3c1可能通过抑制B淋巴祖细胞的糖酵解途径关键基因的表达，控制细胞增殖过程中的能量供应，维持正常淋巴细胞稳态发育。本项目将基于基因敲除动物模型，利用代谢组学分析、Seahorse细胞能量代谢分析、ChIP免疫沉淀、单细胞转录组测序等方法明确Nr3c1通过调控能量代谢在早期B淋巴细胞发育过程中的作用机制，为靶向能量代谢治疗难治、复发ALL提供新思路。

A large amount of clinical evidence as well as preliminary research by our group indicate that the deletion mutation of the glucocorticoid receptor gene (NR3C1) is one of the most important factors that cause drug-resistance and disease relapse of acute lymphoblastic leukemia (ALL). However, the effect of NR3C1 on normal hematopoietic differentiation and its regulatory mechanism are still unclear. In the early stage, our group constructed hematopoietic stem cell conditional Nr3c1 knockout mice and found that the knockout mice had abnormal B lymphocyte development, along with the B lymphoid precursor cells and common lymphoid progenitor cells increased significantly. This phenotype is coincides with the apparent expansion of B lymphoid progenitor cells in human acute B lymphocytes leukemia. Our further preliminary experiments suggest that Nr3c1 may inhibit the expression of key genes in the glycolytic pathway of B lymphoid progenitor cells, control the energy supply during cell proliferation, and maintain the steady-state development of normal lymphocytes. This project will rely on hematopoietic stem cell conditional Nr3c1 knockout mice models, take advantage of metabolomics analysis, Seahorse cell energy metabolism analysis, ChIP immunoprecipitation, single-cell transcriptome sequencing and other methods to clarify the mechanism of Nr3c1 in the early development of B lymphocytes by regulating energy metabolism. This project is expected to provide new ideas for the prevention and treatment of drug-resistance and relapse of acute lymphoblastic leukemia associated with NR3C1 mutation.