Alam P, Tharp D L, Bowles H J, Grisanti L A, Bui H, Bender S B, Bowles D K
Pathology and Integrative Biomedical Sciences, University of Missouri, Columbia, MO, USA.
Dalton Cardiovascular Research Center, University of Missouri, Columbia, MO, USA.
Channels (Austin). 2025 Dec;19(1):2538864. doi: 10.1080/19336950.2025.2538864. Epub 2025 Aug 3.
Increased expression of K3.1 has been found in vascular smooth muscle cells (SMC), macrophages, and T cells in atherosclerotic lesions from humans and mice. Pharmacological inhibition of K3.1 in limiting atherosclerosis has been demonstrated in mice and pigs, however direct, loss-of-function, i.e. gene silencing, studies are absent. Therefore, we generated K3.1Apoe (DKO) mice and assessed lesion development in the brachiocephalic artery (BCA) of DKO versus Apoe mice on a Western diet for 3 months. In BCAs of DKO mice, lesion size and relative stenosis were reduced by ~70% compared to Apoe mice, with no effect on medial or lumen area. Additionally, DKO mice exhibited a significant reduction in macrophage content within plaques compared to Apoe mice, independent of sex. migration assays showed a significant reduction in migration of bone marrow-derived macrophages (BMDMs) from DKO mice compared to those from Apoe mice. experiments using rat aortic smooth muscle cells revealed inhibition of PDGF-BB-induced MCP1/Ccl2 expression upon K3.1 inhibition, while activation of K3.1 further enhanced MCP1/Ccl2 expression. Both and analyses showed that silencing K3.1 had no significant effect on the collagen content of plaque. RNAseq analysis of BCA samples from DKO and Apoe mice revealed PPAR-dependent signaling as a potential key mediator of the reduction in atherosclerosis due to K3.1 silencing. Overall, this study provides the first genetic evidence that K3.1 is a critical regulator of atherosclerotic lesion development and composition and provides novel mechanistic insight into the link between K3.1 and atherosclerosis.
在人类和小鼠动脉粥样硬化病变的血管平滑肌细胞（SMC）、巨噬细胞和T细胞中，已发现K3.1的表达增加。在小鼠和猪中已证明K3.1的药理学抑制作用可限制动脉粥样硬化，但缺乏直接的功能丧失即基因沉默研究。因此，我们构建了K3.1Apoe（双敲除，DKO）小鼠，并评估了DKO小鼠与Apoe小鼠在西方饮食3个月后，其头臂动脉（BCA）中的病变发展情况。在DKO小鼠的BCA中，与Apoe小鼠相比，病变大小和相对狭窄程度降低了约70%，而对中膜或管腔面积没有影响。此外，与Apoe小鼠相比，DKO小鼠斑块内的巨噬细胞含量显著降低，且与性别无关。迁移试验表明，与Apoe小鼠相比，DKO小鼠骨髓来源巨噬细胞（BMDM）的迁移显著减少。使用大鼠主动脉平滑肌细胞的实验表明，抑制K3.1可抑制血小板衍生生长因子-BB（PDGF-BB）诱导的MCP1/Ccl2表达，而激活K3.1则进一步增强MCP1/Ccl2表达。免疫组化和免疫荧光分析均显示，沉默K3.1对斑块的胶原含量没有显著影响。对DKO和Apoe小鼠BCA样本的RNA测序分析表明，PPAR依赖性信号传导是K3.1沉默导致动脉粥样硬化减轻的潜在关键介质。总体而言，本研究提供了首个遗传学证据，证明K3.1是动脉粥样硬化病变发展和组成的关键调节因子，并为K3.1与动脉粥样硬化之间的联系提供了新的机制性见解。
