主控基因RUNX1对银屑病患者骨髓造血干细胞向T细胞分化的影响

We had found that many immuno-character of T cell differentiated from psoriatic hematopoietic stem cell(HSC) were similar to T cell from psoriatic peripheral blood in previous study, and most of the abnormality in Notch signal particpate in the psoriatic differentiation of HSC to T cell. The expression of RUNX1 which play critical role in HSC and differentiation of T cell, was regulated by Notch signal.Recently,our studies indicated that target genes of RUNX1 were abnormal in HSC from psoriasis.The further study of RUNX1 will make the differentiated mechasim of HSC to T cell become clearer in psoriasis.In this study,expression of SCL,methylation of DNA and acetylation of histone will be tested in CD34+ cell. Secondly,the expression or protein methylation of RUNX1 will be interfered.After that the impaction will be invested including protein methylation of RUNX1,interaction between RUNX1 protein and SLC9A3R1, expression of SLC9A3R1 and function of cell adhesion. CD34+ cell will also be differentiated to T cell after interfere, then activity of T cell and expression of RUNX1 target genes be invested. At the same time, the T cell separated from psoriatic peripheral blood will also be tested.The topic is original.

前期研究我们在国际上首先发现银屑病造血干细胞（HSC）分化的T细胞与银屑病T细胞具有许多相似的免疫特性，且Notch信号转导多个环节障碍参与了银屑病HSC向T细胞分化。Notch信号可调节RUNX1表达，而RUNX1是调节HSC和T细胞分化的主控基因，新近我们又得到银屑病HSC RUNX1部分靶基因异常的直接证据。故对RUNX1的深入研究将使银屑病HSC向T细胞分化机理变的更加清楚。本课题拟在前期基础上对银屑病CD34+细胞进行以下研究：①检测 SCL表达、DNA甲基化和组蛋白乙酰化；②转染PRMT1/干扰RUNX1表达，检测转染/干扰对RUNX1蛋白甲基化、与SLC9A3R1蛋白-DNA相互作用、SLC9A3R1表达及细胞粘附的影响；③将转染/干扰后CD34+细胞定向分化为T细胞，检测其对T细胞分化和靶基因表达的影响；④分选外周血T细胞，检测RUNX1靶基因表达等。课题是实质性原创项目。

银屑病是一种在遗传背景下发生的免疫紊乱性皮肤疾病，我们的前期研究结果显示干细胞参与了银屑病的发病。由于项目执行过程中，骨髓取材特别困难，故正如进展报告中所述研究过程中对研究内容进行了部分调整。该项目对银屑病骨髓造血干细胞、骨髓间充质干细胞、皮肤间充质干细胞及外周血T细胞进行了系列研究。间充质干细胞的研究结果显示银屑病患者骨髓和皮肤间充质干细胞分泌细胞因子等活性存在异常，结果证实间充质干细胞在银屑病的发生发展中具有重要作用，为银屑病发病机理的研究提供了新的思路；外周血T细胞研究发现银屑病患者外周血T细胞活性与正常不同，且对表皮通过时间、增殖相关蛋白表达等的影响相似于银屑病皮损，充实了银屑病免疫机理研究的理论依据。项目中对银屑病造血干细胞、间充质干细胞、T细胞等的研究均具有较高的创新性，且取得了很好的研究成果。