Noggin及其共表达lncRNAs调控单核巨噬细胞表型分化及其在动脉硬化血管重塑中的机制与临床研究

Vascular inflammation and calcification are important pathogenesis in plaques stability. Monocytes/macrophages play a major role in the initiation, propagation, and progression of atherosclerosis from a stable to unstable state. With plasticity and heterogeneity, monocytes/macrophages exist in two subsets with different phenotypes and functional properties. M1, the classical activated macrophages, produce pro-inflammatory effects and promote the progression of atherosclerosis. In contrast, M2, the alternative activated macrophages, produce anti-inflammatory effects and may alienate the progression of atherosclerosis. However, microenvironment and precise mechanism in driving monocytes/macrophages differentiation remain to be further elucidated...With the use of whole-blood transcriptome sequencing, we identified an important bone morphogenetic proteins (BPMs) antagonist, noggin (NOG) and a new long non-coding RNA (lncRNA) AC005842.1, represented a significantly higher expression in patients with mixed plaques, compared with those patents with calcification plaques and healthy subjects without coronary plaques who were diagnosed by Coronary Computed Tomography Angiography (CCTA). Next, subcellular localization experiments showed that NOG particularly expressed in macrophages of human carotid atherosclerotic plaques. Furthermore, lncRNA AC005842.1 induced THP-1 derived macrophages polarization by upregulating the expression of NOG. Based on these interesting preliminary findings, we proposed the hypothesis that a feed-forward regulation between lncRNA AC005842.1 and NOG/BMPs signaling could be the key mechanistic basis for the coexistence of chronic inflammation and calcification in monocytes/macrophages in atherosclerosis plaque lesions, which may contribute to vascular remodeling, plaque instability, and atherosclerotic progression...In the present project, we aimed to (1) elucidate whether AC005842.1 promoted NOG/BMPs signaling in macrophages and modulate macrophages’ polarization and functions during the development of atherosclerosis, via the cell model of THP-1 derived macrophages infected with NOG lentivirus or siRNA and the ApoE-/- mice model of atherosclerosis with monocytes-specific knockdown of NOG-/- and AC005842.1-/-; (2) investigate the relationship between circulating NOG/AC005842.1 levels and progression of coronary artery calcification and the risk of future cardiovascular diseases during the follow-up in an established, perspective coronary heart disease cohort.

前期工作通过全转录组测序发现，与无斑块对照和斑块完全钙化组比较，钙化拮抗剂--头蛋白（NOG）及其共表达的lncRNA分子AC005842.1在冠脉混合斑块组患者高表达；亚细胞定位在斑块部位单核/巨噬细胞，并发现AC005842.1促进NOG表达及其下游骨形态发生蛋白（BMPs）信号通路，提示AC005842.1可能通过NOG调控巨噬细胞表型分化、功能，影响血管钙化和斑块进展。本项目拟解决问题1：通过基因过表达和siRNA阻断技术，以及单核细胞特异敲除NOG-/-、AC005842.1-/-的ApoE-/-动脉粥样硬化小鼠模型，明确AC005842.1是否通过NOG及下游BMPs信号调控巨噬细胞活动，影响斑块进展和炎性重塑；2：检测冠脉钙化前瞻性队列外周血AC005842.1和NOG与钙化积分和心血管事件的关系，将为阐明lncRNAs调控单核/巨噬细胞分化及血管炎性重塑的机制提供新靶点。

动脉粥样硬化是心脑血管疾病发生和发展的重要病理基础，循环血中的单核细胞迁移并黏附于活化的血管内皮细胞，浸润至内膜下层形成巨噬细胞是动脉粥样硬化的起始环节，单核-巨噬细胞通过表型转换和分泌炎症因子等机制调控动脉粥样硬化发展的各个阶段。斑块的稳定性决定了动脉粥样硬化的危险程度。本研究采用外周血全转录组测序技术，首次鉴别出一种lncRNA AC005842.1分子在具有混合易损斑块的冠心病患者外周血中特异高表达。并通过离体细胞实验、基因过表达和基因功能siRNA阻断技术、以及基因过表达和敲除的动脉粥样硬化小鼠模型等动物实验，发现lncRNA AC005842.1在调控单核-巨噬细胞活性、影响斑块不稳定性中发挥着重要作用。该lncRNA分子 位于人12号染色体，长686bp，我们将其命名为lncRNA APAT。本项目研究结果提示：lncRNA APAT具有主动脉粥样硬化斑块形成的作用，有望作为新的标记物对动脉粥样硬化斑块稳定性和进展进行评估预测，为基因治疗、药物治疗等临床应用提供了治疗靶点和重要依据。