新的嗅觉受体突变在再生障碍性贫血骨髓造血微环境损伤中的作用以及补肾健脾祛瘀法的修复机制

我们前期通过全基因组测序发现了再障MSCs中普遍存在新的嗅觉受体OR56B1突变。本项目将探讨OR56B1突变对BMSCs功能，造血干祖细胞维持，造血微环境和免疫环境的影响以及补肾健脾祛瘀法对OR56B1突变的修复作用。项目将通过CCK-8法、Annexin V-FITC/PI双染法、western-blot、ELISA、实时荧光定量PCR、Transwell、RNA干扰、CFSE染色追踪、免疫荧光染色、基因定点突变等技术，来明确以下情况，包括OR56B1受体在BMSCs上天然表达和作用；OR56B1受体点突变对BMSCss影响；OR56B1突变对Dicer的影响；OR56B1受体点突变对正常造血干细胞的支持作用；OR56B1受体点突变对免疫微环境的调控作用以及补肾健脾祛瘀法对上述损伤的修复作用。通过该研究可以深入了解造血微环境在再障发病中的作用，借此寻找新的再障预后判断和治疗靶点。

We found a new olfactory receptor mutation, OR56B1, in BMSCs with whole genome sequencing.This project will explore the injury role of OR56B1 mutation to the function of BMSCs,to the maintenance of hematopoietic and stem progenitor cells,and to hematopoietic microenvironment and immune environment,and the repair mechanism of “notifying kidney and spleen and removing stasis”.The products were identified by CCK-8, Annexin V-FITC / PI double staining, western-blot, ELISA, real-time fluorescence quantitative PCR, Transwell, RNA interference, CFSE staining, immunofluorescence staining and gene site mutation,to clarify the following cases,including OR56B1 receptor in BMSCs on the natural expression and role;effects of OR56B1 receptor point mutation on BMSCs ;effects of OR56B1 Mutation on Dicer；the support of OR56B1 receptor point mutation to normal hematopoietic stem cells,the role of OR56B1 receptor mutations to the regulation of immune microenvironment and the effect of “notifying kidney and spleen and removing stasis”on the repair of these lesions.Through the study will let us to understand the role of BMHM on the pathogenesis of AA but also to find a new prognosis and therapeutic target of AA.