Knockout of the NONO Gene Inhibits Neointima Formation in a Mouse Model of Vascular Injury
Knockout of the NONO Gene Inhibits Neointima Formation in a Mouse Model of Vascular Injury
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NONO 基因的敲除抑制血管损伤小鼠模型中新内膜的形成
DOI：
10.1161/atvbaha.119.313581
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发表时间：
2021
期刊：
影响因子：
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通讯作者：
Zhang Cheng
中科院分区：
文献类型：
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作者：
Xu Xingli;Xu Xinghua;Mao Yang;Lu Lin;Ma Jing;Zheng Tengfei;Zhang Jie;Zhang Meng;Meng Linlin;Ma Lianyue;Cheng Jing;Chen Wenqiang;Jiang Hong;Zhang Yun;Zhang Cheng
NONO（non-POU domain-containing octamer-binding protein）是一种多功能核蛋白，参与RNA合成、转录调控和DNA修复。然而，NONO对血管损伤或再狭窄引起的新生内膜的影响尚不清楚。我们假设NONO是维持血管完整性所必需的，NONO敲除可能抑制新生内膜形成。方法和结果NONOgene KO（敲除; NONO KO或NONOgt/0）小鼠使用CRISPR/Cas9（成簇规则间隔短回文重复序列/成簇规则间隔短回文重复序列相关9）技术从C57 BL/6 J小鼠产生。结扎小鼠左颈总动脉，诱导颈动脉新生内膜形成。原代培养的小鼠主动脉血管平滑肌细胞（VSMCs）来源于野生型和NONOgt/0小鼠腹主动脉的培养液。人冠状动脉含有动脉粥样硬化斑块和正常冠状动脉来自身体供体。组织学染色显示NONO在人冠状动脉粥样硬化病变和小鼠颈动脉结扎中表达增加。NONO主要来源于新生内膜的VSMCs。NONO缺乏小鼠的颈动脉结构与对照小鼠无显著差异。然而，颈动脉结扎后，NONOgt/0小鼠显示结扎动脉中新生内膜厚度减少。NONO缺乏导致VSMCs增殖和迁移减少，新生内膜和VSMCs收缩标志基因表达增加。NONO与VSMCs中的Erk（胞外调节激酶）1/2相互作用，影响其在VSMCs中的活化，提示Erk信号级联可能介导NONO在VSMCs中的作用。结论NONO不是维持血管完整性所必需的，但NONO基因敲除可逆转VSMCs增殖、迁移和表型转换的病理过程。这些作用的机制涉及NONO和Erk信号级联的相互作用。因此，抑制NONO可能为内膜增厚相关的心血管疾病提供一种新的治疗策略。
ObjectiveThe NONO (non-POU domain-containing octamer-binding protein) is a multifunctional nuclear protein involved in RNA synthesis, transcriptional regulation, and DNA repair. However, the effect of NONO on neointima induced by vascular injury or restenosis remains unclear. We hypothesized that NONO is required for maintaining vascular integrity and NONO knockout may inhibit neointima formation.Approach and ResultsNONOgene KO (knockout; NONO KO or NONOgt/0) mice were produced from C57BL/6J mice using the CRISPR/Cas9 (clustered regularly interspaced short palindromic repeats/clustered regularly interspaced short palindromic repeat–associated 9) technique. The left common carotid artery of mice was ligated to induce carotid neointima formation. The primary mouse aortic vascular smooth muscle cells (VSMCs) were derived from the media of wide-type and NONOgt/0mouse aortas for in vitro studies. Human coronary arteries containing atherosclerotic plaque and normal coronary arteries were obtained from body donors. Histological staining demonstrated that NONO expression was increased in human coronary atherosclerotic lesions and mouse ligated carotid arteries. Moreover, the increased NONO was primarily from VSMCs of neointima. Mice with NONO deficiency showed no significant difference in carotid artery structure from control mice. However, after carotid artery ligation, NONOgt/0mice exhibited reduced neointima thickness in ligated arteries. NONO deficiency led to decreased proliferation and migration of VSMCs and increased expression of contractile marker genes in neointima and VSMCs. The mechanistic study indicated that NONO interacted with Erk (extracellular regulated kinase) 1/2 in VSMCs and affected its activation in VSMCs, implying Erk signaling cascade might mediate the roles of NONO in VSMCs.ConclusionsNONO was not required for maintaining vascular integrity, but NONO knockout reversed the pathological processes mediated by increased proliferation, migration, and phenotypic switching of VSMCs. The mechanism of these effects involved an interaction of NONO and Erk signaling cascade. Thus, inhibition of NONO may provide a novel therapeutic strategy in cardiovascular disease associated with intimal thickening.