PARP-1编码区错义变异rs1136410调节单核-巨噬细胞功能降低冠心病发病风险的分子机制

Poly(ADP-ribose) polymerase 1 (PARP-1) gene regulates transcription, DNA repair, and cell metabolism, leading to endothelial dysfunction. However, the exact roles of PARP-1 in monocyte-macrophage functions are largely unknown. We previously showed that rs1136410, a missense variant in PARP-1, protected against the risk and severity of coronary artery disease (CAD), and was inversely correlated with PARP-1 activities in peripheral blood mononuclear cells. Bioinformatics suggested a potential impact of rs1136410 on the binding between PARP-1 and a transcription factor, GATA2. Thus, we assume that rs1136410 modified PARP-1 expression and enzyme activity to decrease CAD risk. To unveil the detailed mechanism, we propose the following expeiments: 1) to assess the impacts of rs1136410 on expression, enzyme activitiy, and stability of PARP-1 in monocyte-macrophage cells; 2) to assess whether rs1136410 affects the binding between PARP-1 and GATA2, and the transcriptional activity of PARP-1; 3) to comapre the differences in polarization, inflammation, cholesterol efflux, and adhesion to endothelium between different genotypes of rs11364410 in monocyte-macrophage cells; 4) to reveal the influence of rs1136410 knock-in on atherosclerosis development and macrophage functions in a mouse model.

聚腺苷酸二磷酸核糖转移酶1（PARP-1）基因调控转录、DNA修复和细胞代谢，致内皮功能紊乱。然而，PARP-1对单核-巨噬细胞功能的影响尚未阐明。我们前期发现PARP-1编码区错义变异rs1136410是冠心病发病风险和严重程度的保护因素，并与单个核细胞内PARP-1活性负相关。生物信息学分析显示rs1136410潜在影响PARP-1基因和转录因子GATA2的结合。由此假设rs1136410调控PARP-1表达和酶活性降低冠心病风险。为探索具体机制，拟分析：1.rs1136410对单核-巨噬细胞内PARP-1表达、酶活性和稳定性的影响；2.rs1136410对PARP-1基因和GATA2的结合，及该结合对PARP-1转录活性的作用；3.rs1136410对单核-巨噬细胞极化、炎症、胆固醇转运及粘附内皮功能的影响；4.rs1136410敲入对小鼠动脉粥样硬化进展和巨噬细胞功能的作用。

聚腺苷酸二磷酸核糖转移酶1（PARP-1）基因调控转录、DNA修复和细胞代谢，致内皮功能紊乱。然而，PARP-1对单核-巨噬细胞功能的影响尚未阐明。我们前期发现PARP-1编码区错义变异rs1136410与冠心病风险降低相关。为阐明其作用机制，本项目以THP-1单核细胞系、人原代单核-巨噬细胞、基因敲入小鼠模型和冠心病队列人群为研究对象，采用分子生物学干预、表型观察、流行病调查等技术手段，获得如下科学进展：1）分子机制上，rs1136410所在区域高度保守，其突变等位的出现改变了单核-巨噬细胞内PARP-1 mRNA稳定性，抑制转录因子GATA2结合PARP-1转录调控区域，进而下调PARP-1转录活性和表达；2）细胞功能上，rs1136410通过调控PARP-1表达减少人原代单核-巨噬细胞粘附内皮，抑制巨噬细胞向M1促炎型极化；3）动物实验显示，rs1136410敲入可保护高脂饮食小鼠的动脉和心脏功能，减少斑块内巨噬细胞数量，并降低斑块面积；4）人群中，基于包括rs1136410在内的多个遗传位点建立孟德尔随机化模型，证实PARP-1过度活化与冠心病风险升高之间的因果关系。本项目首次对单核-巨噬细胞中PARP-1编码区错义突变的功能和调控机制进行了较为系统、深入的研究，为冠心病的分子遗传学病因提供了新证据。项目资助成果已在ATVB、Cardivasc Diabetol等期刊发表SCI论文6篇。项目组成员继续获得国家自然科学基金面上项目1项、青年项目2项。

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