CARMN Loss Regulates Smooth Muscle Cells and Accelerates Atherosclerosis in Mice.
CARMN Loss Regulates Smooth Muscle Cells and Accelerates Atherosclerosis in Mice.
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DOI：
10.1161/circresaha.120.318688
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发表时间：
2021-04-30
影响因子：
20.1
通讯作者：
Baker AH
中科院分区：
文献类型：
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作者：
Vacante F;Rodor J;Lalwani MK;Mahmoud AD;Bennett M;De Pace AL;Miller E;Van Kuijk K;de Bruijn J;Gijbels M;Williams TC;Clark MB;Scanlon JP;Doran AC;Montgomery R;Newby DE;Giacca M;O'Carroll D;Hadoke PWF;Denby L;Sluimer JC;Baker AH
在动脉粥样硬化病变的微环境中，血管平滑肌细胞（vSMC）转换为去分化状态，但驱动这种转换的潜在分子机制尚未完全了解。长链非编码RNA（lncRNA）在血管病理过程中失调，但对它们参与控制vSMC功能知之甚少。CARMN是一种位于microRNA-143和-145（miR-143和miR-145）上游的lncRNA，两者都参与vSMC功能。我们研究了独立于miR-143和miR-145的lncRNA CARMN作为体外vSMC表型的潜在调节剂的作用，以及在体内动脉粥样硬化发展过程中其损失的后果。我们假设CARMN的丧失是控制向致动脉粥样硬化vSMC表型的功能转换的主要事件，并加速了体内斑块的发展。在人冠状动脉平滑肌细胞（hCASMC）中使用GapmeR沉默CARMN lncRNA的表达，揭示了GapmeR介导的CARMN丧失对miR-143和miR-145 miRNA表达产生负面影响。CARMN缺失的hCASMCs的RNA测序显示大的转录组学变化，与vSMC增殖、迁移、炎症、脂质代谢和去分化相关。miR-143和miR-145模拟物的使用揭示了CARMN以miRNA非依赖性方式调节hCASMC增殖。在人类和小鼠中，CARMN和相关miRNA在晚期与早期动脉粥样硬化病变中下调。使用CRISPR（成簇规则间隔短回文重复序列）- Cas9（CRISPR相关蛋白9）敲除方法，我们探索了体内动脉粥样硬化期间CARMN消耗的影响。与体外结果一致，CARMN的敲除在稳态条件下损害了miR-143和miR-145的表达。重要的是，当在这些小鼠中诱导动脉粥样硬化时，CARMN敲除增加了体积，大小，促炎LGALS 3表达细胞含量和改变斑块组成，产生了晚期表型。我们将CARMN lncRNA的早期丢失确定为关键事件，其在体外使vSMC朝向促动脉粥样硬化表型并加速体内动脉粥样硬化的发展。
In the microenvironment of atherosclerotic lesions, vascular smooth muscle cells (vSMCs) switch to a dedifferentiated state but the underlying molecular mechanisms driving this switch are not fully understood. Long noncoding RNAs (lncRNAs) are dysregulated during vascular pathology, but relatively little is known about their involvement in controlling vSMCs function. CARMN is a lncRNA located immediately upstream of the microRNAs-143 and -145 (miR-143 and miR-145), both involved in vSMCs function. We investigated the role of the lncRNA CARMN, independent from miR-143 and miR-145, as potential a regulator of vSMC phenotypes in vitro and the consequences of its loss during the development of atherosclerosis in vivo. We hypothesized that loss of CARMN is a primary event controlling the functional switch towards pro-atherogenic vSMC phenotype and accelerates the development of the plaques in vivo. Expression of CARMN lncRNA was silenced using GapmeRs in human coronary arterial smooth muscle cells (hCASMCs), revealing that GapmeR-mediated loss of CARMN negatively affects miR-143 and miR-145 miRNA expression. RNA sequencing of CARMN-depleted hCASMCs revealed large transcriptomic changes, associated with vSMC proliferation, migration, inflammation, lipid metabolism and dedifferentiation. The use of miR-143 and miR-145 mimics revealed that CARMN regulates hCASMC proliferation in a miRNA-independent manner. In human and mouse, CARMN and associated miRNAs were downregulated in advanced versus early atherosclerotic lesions. Using a CRISPR (clustered regularly interspaced short palindromic repeats)- Cas9 (CRISPR-associated protein 9) knock-out approach, we explored the implications of CARMN depletion during atherosclerosis in vivo. Consistent with in vitro results, the knock-out of CARMN impaired the expression of miR-143 and miR-145 under homeostatic conditions. Importantly, when atherosclerosis was induced in these mice, CARMN knock-out increased the volume, size, pro-inflammatory LGALS3-expressing cells content and altered plaque composition, yielding an advanced phenotype. We identified the early loss of CARMN lncRNA as critical event which primes vSMCs towards a pro-atherogenic phenotype in vitro and accelerates the development of atherosclerosis in vivo.