Lnc-072240靶向TIM-4调控巨噬细胞炎症反应在深静脉血栓形成中的作用与机制研究

巨噬细胞介导的炎症反应是导致深静脉血栓（DVT）形成的重要因素，但其调控机制尚不清楚。TIM-4是负调控巨噬细胞炎症反应的重要免疫分子，我们前期发现敲除TIM-4促进巨噬细胞炎症反应及血栓形成，并发现TIM-4在DVT患者外周血及DVT小鼠血栓中巨噬细胞表达显著降低，结合全转录组高通量筛选发现长链非编码lnc-072240与TIM-4的关键负调控因子miR-93-5p具有潜在结合位点，且在DVT患者表达显著降低，与TIM-4表达呈显著正相关。基于以上工作基础，推测lnc-072240靶向TIM-4调控巨噬细胞炎症反应在DVT形成中发挥了重要作用。本项目拟采用Tim-4-/-小鼠及CRISRP/Cas9等技术，探讨lnc-072240作为竞争性内源RNA靶向TIM-4调控巨噬细胞炎症反应在DVT形成中的作用与机制，以期从非编码RNA免疫调控角度阐释DVT发病机理，为治疗提供新的靶点与思路。

Macrophage inflammatory responses are the critical steps for deep vein thrombosis（DVT）progression, but molecular mechanism underlying the pathogenesis of DVT remains unclear so far. T-cell immunoglobulin domain and mucin domain 4（TIM-4）is a key regulator in suppressing macrophage inflammatory responses. Our preliminary data demonstrated that TIM-4 deficiency in macrophages promoted inflammatory responses and thrombosis. Additionally, the expression of TIM-4 was decreased in both peripheral blood mononuclear cells（PBMCs）from DVT patients and macrophages from thrombus tissue of DVT mice. These findings, together with high-throughput full transcriptome sequencing aimed to identify key factors regulating the abnormal expression of TIM-4, revealed that miR-93-5p suppressed expression of TIM-4, which possesses the potential binding site with TIM-4. Subsequent studies also disclosed that lnc-072240 possessed the potential binding site with miR-93-5p, and the expression of lnc-072240 was decreased in PBMCs from DVT patients and showed positive correlation with TIM-4. These exciting preliminary findings led us to hypothesize that lnc-072240 plays a critical role in regulating macrophage inflammatory responses in the pathogenesis of DVT by regulating TIM-4. In brief, we will leverage our Tim-4 knockout mice and CRISPR/Cas9 technique to further explore the functions and mechanisms of lnc-072240 as a competing endogenous RNA（ceRNA）regulating TIM-4-mediated macrophage inflammatory responses in DVT. The proposed studies will not only provide new insights into the pathogenesis of DVT from aspect of epigenetic modification of non-coding RNA, but may also offer new intervention targets for treating DVT in clinic.