GATA-1/miR-150/MYB通路调控低危MDS巨核细胞分化的机制及干预研究

The main clinical feature of low-risk MDS is refractory cytopenia. It needs long term and expensive supportive care, which brings heavy burdens to patients and society. Therefore, its treatment has become a research hotspot. We screen out amifostine (AMF) as a potential drug treating low-risk MDS by bioinformatics. Clinical study found that AMF could improve dysmegakaryocytopoiesis and increase the number of blood cells of low-risk MDS, but the mechanism is still not clear. Our previous study found that AMF could induce Dami cell differentiation, which is accompanied with an increased nuclear translocation of GATA-1 and miR-150 expression. Bioinformatics found that amifostine may promote megakaryocyte differentiation in low-risk MDS by GATA-1/miR-150/MYB pathway. In this study, we will figure out the role of this pathway on amifostine's promoting differentiation of Dami cells, and verify the relationship between GATA-1/miR-150/MYB pathway and dysmegakaryocytopoiesis of low-risk MDS on human primary cells. Finally, we will comprehensively analyze the relationship between clinical data and expression of GATA-1, miR-150, and MYB of low-risk MDS bone marrow cells. And we will find out a molecular marker to predict the efficacy of AMF in low-risk MDS, which could provide a theoretical basis for clinical stratified treatment of MDS.

低危MDS主要临床特征是难治性血细胞减少，需要长期昂贵的支持治疗，给患者和社会带来沉重负担，其治疗成为研究热点。课题组利用生物信息学手段筛选出治疗低危MDS的药物依硫磷酸（AMF），临床研究发现AMF能改善低危MDS巨核细胞分化障碍，提高血小板数量，但机理不明。我们前期研究发现AMF能诱导巨核细胞系Dami分化，此过程伴随转录因子GATA-1入核增加及miR-150表达升高，生物信息学分析提示AMF治疗低危MDS可能和GATA-1/miR-150/MYB通路有关。本研究拟用细胞系验证AMF通过GATA-1/miR-150/MYB通路促进巨核细胞分化，用脐血和骨髓原代细胞验证此通路和低危MDS巨核细胞分化障碍的关系，最终将低危MDS骨髓GATA-1、miR-150、MYB的表达水平结合临床数据综合分析，找出预测AMF治疗低危MDS疗效的分子标记，为临床MDS的分层治疗提供理论基础。

低危MDS主要临床特征是难治性血细胞减少，对常规促进造血的药物反应差。课题组前期临床研究发现依硫磷酸（AMF）能改善低危MDS巨核细胞分化障碍，升高血小板数量，但机理不明。本研究发现AMF可以抑制HEL细胞的增殖并促进HEL细胞向成熟巨核细胞分化。AMF刺激HEL细胞后，发现HEL细胞体积增大，细胞内线粒体数量增多，DNA倍体化增加，并且巨核细胞相关特征性标记CD42的mRNA和蛋白水平均升高。进一步检测HEL细胞活性氧水平，发现AMF可能通过诱导活性氧产生促进HEL细胞向巨核细胞分化。通过60Coγ射线照射C57BL/6J小鼠制造骨髓损伤模型，发现AMF可以促进骨髓损伤小鼠的骨髓重建及外周血血小板计数恢复。本课题发现AMF能通过影响能量代谢促进巨核细胞分化，为低危MDS提供了一种新的潜在治疗策略。本课题发表论文3篇，培养硕士研究生2名，其中1名已经取得硕士学位，1名在读。课题各项支出与预算基本相符，剩余经费计划用于本项目后续研究支出。