Department of Ultrasound, The Second Affiliated Hospital of Harbin Medical University, Harbin, China; The Key Laboratory of Myocardial Ischemia, Harbin Medical University, Ministry of Education, Harbin, Heilongjiang, China.
Department of Cardiology, The First Affiliated Hospital of Harbin Medical University, Harbin, China; Department of Cardiology, Guangdong Provincial People's Hospital, Guangdong Academy of Medical Sciences, Guangzhou, China.
Int J Biochem Cell Biol. 2022 Oct;151:106294. doi: 10.1016/j.biocel.2022.106294. Epub 2022 Aug 27.
Angiogenesis plays a key role in coronary collateral circulation (CCC), the compensatory formation of new blood vessels during chronic total coronary occlusion. This study aimed to determine whether plasmacytoma variant translocation 1 (PVT1), a long non-coding (lnc) RNA involved in tumor angiogenesis, plays a role in regulating angiogenesis during chronic coronary ischemia.
Patients with coronary artery disease, and ≥ 90% stenosis, were examined and divided into "Good" and "Poor" CCC groups based on Rentrop Cohen classification. RNA samples were obtained from all patients, as well as from oxygen and glucose-deprived (OGD) HUVECs. PVT1, miR-15b-5p and AKT3 levels were measured with RT-qPCR or Western blot, while HUVEC migration and angiogenesis were detected by, respectively, wound-healing and tube formation assays. Luciferase reporter assay confirmed direct PVT1-miR-15b-5p binding.
Increased PVT1 was found in "Good CCC" patient plasma, along with being highly expressed among OGD HUVECs; PVT1 knockdown reduced HUVEC migration, tube formation, and pro-angiogenic factor expression. Conversely, OGD HUVECs had downregulated miR-15b-5p, and miR-15b-5p overexpression significantly depressed their angiogenic capabilities. These PVT1 knockdown- or miR-15b-5p overexpression-associated reductions in angiogenic effects were reversed by AKT3 overexpression. In vivo, neovascularization and functioning in both ischemic mice hind-limbs and infarcted myocardium injected with ADV-sh-PVT1 were reduced, which were ameliorated by concurrent antagomiR-15b-5p injections.
Circulating PVT1 may serve as a useful biomarker to distinguish between good versus poor CCC, as it is involved in orchestrating angiogenesis via the miR-15b-5p-AKT3 axis; it thus has potential as a target for treating ischemic disease.
血管生成在冠状动脉侧支循环（CCC）中起着关键作用，即在慢性全冠状动脉闭塞期间新血管的代偿形成。本研究旨在确定浆细胞瘤变异易位 1（PVT1），一种参与肿瘤血管生成的长链非编码（lnc）RNA，是否在慢性冠状动脉缺血期间调节血管生成中发挥作用。
对患有冠心病且≥90%狭窄的患者进行检查，并根据 Rentrop Cohen 分类将其分为“良好”和“不良” CCC 组。从所有患者以及缺氧和葡萄糖剥夺（OGD）的 HUVEC 中获取 RNA 样本。通过 RT-qPCR 或 Western blot 测量 PVT1、miR-15b-5p 和 AKT3 水平，通过划痕愈合和管形成测定分别检测 HUVEC 迁移和血管生成。荧光素酶报告实验证实了 PVT1-miR-15b-5p 的直接结合。
在“良好 CCC”患者的血浆中发现 PVT1 增加，并且在 OGD 的 HUVEC 中高度表达；PVT1 敲低减少了 HUVEC 的迁移、管形成和促血管生成因子的表达。相反，OGD 的 HUVEC 下调了 miR-15b-5p，而 miR-15b-5p 的过表达显著抑制了它们的血管生成能力。这些 PVT1 敲低或 miR-15b-5p 过表达相关的血管生成效应的降低可通过 AKT3 的过表达逆转。在体内，用 ADV-sh-PVT1 注射缺血小鼠后肢和梗死心肌的新生血管和功能减少，用同时注射 antagomiR-15b-5p 减轻。
循环 PVT1 可作为区分良好与不良 CCC 的有用生物标志物，因为它通过 miR-15b-5p-AKT3 轴来协调血管生成；因此，它具有作为治疗缺血性疾病的靶点的潜力。
