Ali Ashrifa, Roy Bhaskar, Schott Micah B, Grove Bryon D
J Vasc Res. 2025 Jul 18:1-24. doi: 10.1159/000547350.
This study investigates the role of AKAP12 in endothelial cell motility with a specific focus on AKAP12 variants, AKAP12v1 and AKAP12v2. Previous work has shown that AKAP12, a multivalent A-kinase anchoring protein that binds to PKA and several other proteins regulating protein phosphorylation, is expressed at low levels in most endothelia in vivo but is expressed at higher levels in cells in vitro. Here, we found that AKAP12 expression in endothelial cell (HUVEC) cultures was cell density-dependent, with the expression being highest in subconfluent cultures and lowest in confluent cultures. AKAP12 expression was also elevated in cells at the wound edge of wounded endothelial cell monolayers. Knockdown of variants 1 and 2 inhibited cell migration. However, CRISPR/Cas9 knockout of AKAP12v1 enhanced migration, indicating that the absence of this variant and the presence of AKAP12v2 likely alters the signaling events controlling cell motility. Further analysis using bulk RNA sequencing revealed that the loss of AKAP12v1 affected genes associated with cell migration and intercellular junctions. We propose that AKAP12v1 and AKAP12v2 play distinct yet complementary roles in endothelial cell migration and likely work together in controlling the signaling events associated with vascular repair and development.
本研究调查了AKAP12在内皮细胞迁移中的作用，特别关注AKAP12变体AKAP12v1和AKAP12v2。先前的研究表明，AKAP12是一种多价A激酶锚定蛋白，可与PKA和其他几种调节蛋白磷酸化的蛋白结合，在体内大多数内皮细胞中低水平表达，但在体外细胞中高水平表达。在此，我们发现内皮细胞（HUVEC）培养物中AKAP12的表达呈细胞密度依赖性，在亚汇合培养物中表达最高，在汇合培养物中最低。在受伤内皮细胞单层的伤口边缘细胞中，AKAP12表达也升高。变体1和2的敲低抑制了细胞迁移。然而，AKAP12v1的CRISPR/Cas9敲除增强了迁移，表明该变体的缺失和AKAP12v2的存在可能改变了控制细胞运动的信号事件。使用批量RNA测序的进一步分析表明，AKAP12v1的缺失影响了与细胞迁移和细胞间连接相关的基因。我们提出，AKAP12v1和AKAP12v2在内皮细胞迁移中发挥不同但互补的作用，并可能共同控制与血管修复和发育相关的信号事件。
