Nanjing Comprehensive Stroke Center, Affiliated Nanjing Brain Hospital, Nanjing Medical University, Nanjing, PR China.
Nanjing Comprehensive Stroke Center, Affiliated Nanjing Brain Hospital, Nanjing Medical University, Nanjing, PR China.
World Neurosurg. 2022 Feb;158:e880-e888. doi: 10.1016/j.wneu.2021.11.104. Epub 2021 Nov 27.
Moyamoya disease (MMD) is an idiopathic cerebrovascular disease with unknown etiology. Long noncoding RNA (lncRNA) and messenger RNA (mRNA) profiles in MMD remain unknown. In this current study, we aim to investigate lncRNA-mRNA co-expression pattern and their biological functions in superficial temporal artery (STA) of MMD.
STA of 3 MMD patients without RNF213 mutation and 3 age-matched controls were obtained for transcriptomic RNA sequencing. Bioinformatics analysis was performed to investigate their molecular functions and interactions. Then, differentially expressed genes relative to vascular remodeling were further validated by quantitative real-time polymerase chain reaction and immunofluorescence. WNT5A functions were tested by tube formation assay and wound scratching assay in human microvascular endothelial cells (HMECs).
We detected 6235 different lncRNAs and 2065 different mRNAs from the RNA-sequencing between MMD patients and controls (P < 0.05; fold change >2.0). Gene ontology showed that altered mRNAs were enriched for endothelial cell morphogenesis and positive regulation of angiogenesis, which were closely related with vascular remodeling. We then searched 76 altered genes related with vascular remodeling and applied Kyoto Encyclopedia of Genes and Genomes analysis. Integrated analysis of lncRNA-TF-mRNA co-expression networks and gene verifications indicated that molecular including WNT5A, TEK, and GATA2 may contribute to the vascular malformation of MMD. Overexpression of WNT5A in HMECs promoted tube formation and cell migration.
In MMD patients, genes related to vascular remodeling including WNT5A and their regulators were aberrantly disrupted. These results will help elucidate the complicated pathogenic mechanism of MMD and develop potential therapeutic targets facilitating MMD angiogenesis in the future.
烟雾病（MMD）是一种病因不明的特发性脑血管病。MMD 的长链非编码 RNA（lncRNA）和信使 RNA（mRNA）谱尚不清楚。在本研究中，我们旨在研究 MMD 患者颞浅动脉（STA）中的 lncRNA-mRNA 共表达模式及其生物学功能。
从 3 名无 RNF213 突变的 MMD 患者和 3 名年龄匹配的对照者的 STA 中获取转录组 RNA 测序。进行生物信息学分析以研究它们的分子功能和相互作用。然后，通过定量实时聚合酶链反应和免疫荧光进一步验证与血管重塑相关的差异表达基因。在人微血管内皮细胞（HMEC）中通过管形成测定和划痕实验测试 WNT5A 的功能。
我们从 MMD 患者和对照组之间的 RNA-seq 中检测到 6235 个不同的 lncRNA 和 2065 个不同的 mRNA（P<0.05；倍数变化>2.0）。基因本体论显示，改变的 mRNA 富集于内皮细胞形态发生和血管生成的正调控，这与血管重塑密切相关。然后，我们搜索了 76 个与血管重塑相关的改变基因，并进行了京都基因与基因组百科全书分析。lncRNA-TF-mRNA 共表达网络和基因验证的综合分析表明，包括 WNT5A、TEK 和 GATA2 在内的分子可能有助于 MMD 的血管畸形。在 HMEC 中过表达 WNT5A 可促进管形成和细胞迁移。
在 MMD 患者中，与血管重塑相关的基因，包括 WNT5A 及其调节因子，发生异常破坏。这些结果将有助于阐明 MMD 的复杂发病机制，并为未来促进 MMD 血管生成开发潜在的治疗靶点。
