Department of Neurosurgery, Beijing Tiantan Hospital, Capital Medical University, Beijing, China.
China National Clinical Research Center for Neurological Diseases, Beijing, China.
CNS Neurosci Ther. 2023 May;29(5):1312-1324. doi: 10.1111/cns.14103. Epub 2023 Jan 31.
To explore the underlying mechanism by which low-frequency KRAS mutations result in extensive EndMT occurrence.
Exosomes derived from primarily cultured brain arteriovenous malformation (bAVMs) and human umbilical vein endothelial cells (HUVECs) transfected with KRAS , KRAS , or KRAS lentiviruses were isolated, and their effects on HUVECs were identified by western blotting and immunofluorescence staining. The expression levels of exosomal microRNAs (miRNAs) were evaluated by miRNA microarray, followed by functional experiments on miR-3131 and detection of its downstream target, and miR-3131 inhibitor in reversing the EndMT process induced by KRAS -transfected HUVECs and bAVM endothelial cells (ECs) were explored.
Exosomes derived from KRAS bAVM ECs and KRAS -transfected HUVECs promoted EndMT in HUVECs. MiR-3131 levels were highest in the exosomes of KRAS -transfected HUVECs, and HUVECs transfected with the miR-3131 mimic acquired mesenchymal phenotypes. RNA-seq and dual-luciferase reporter assays revealed that PICK1 is the direct downstream target of miR-3131. Exosomal miR-3131 was highly expressed in KRAS bAVM compared with non-KRAS-mutant bAVM or HUVEC . Finally, a miR-3131 inhibitor reversed EndMT in HUVECs treated with exosomes or the supernatant of KRAS -transfected HUVECs and KRAS bAVM ECs.
Exosomal miR-3131 promotes EndMT in KRAS-mutant bAVMs, and miR-3131 might be a potential biomarker and therapeutic target in KRAS -mutant bAVMs.
探讨低频 KRAS 突变导致广泛 EndMT 发生的潜在机制。
分离源自原代培养的脑动静脉畸形（bAVM）和转染 KRAS G12V、KRAS G12D 或 KRAS G13D 的人脐静脉内皮细胞（HUVEC）的外泌体，并通过 Western blot 和免疫荧光染色鉴定其对 HUVEC 的作用。通过 miRNA 微阵列评估外泌体 microRNAs（miRNAs）的表达水平，随后对 miR-3131 及其下游靶基因进行功能实验，并检测 miR-3131 抑制剂在逆转 KRAS 转染的 HUVEC 和 bAVM 内皮细胞（EC）诱导的 EndMT 过程中的作用。
源自 KRAS G12V bAVM EC 和 KRAS G12V 转染的 HUVEC 的外泌体促进了 HUVEC 的 EndMT。KRAS G12V 转染的 HUVEC 的外泌体中 miR-3131 水平最高，转染 miR-3131 模拟物的 HUVEC 获得间充质表型。RNA-seq 和双荧光素酶报告基因检测显示 PICK1 是 miR-3131 的直接下游靶基因。与非 KRAS 突变型 bAVM 或 HUVEC 相比，KRAS G12V bAVM 中的外泌体 miR-3131 表达水平较高。最后，miR-3131 抑制剂逆转了用外泌体或 KRAS G12V 转染的 HUVEC 和 KRAS G12V bAVM EC 的上清液处理的 HUVEC 中的 EndMT。
外泌体 miR-3131 促进 KRAS 突变型 bAVM 中的 EndMT，miR-3131 可能是 KRAS 突变型 bAVM 的潜在生物标志物和治疗靶点。
