Exosomal miR-27a-3p derived from tumor-associated macrophage suppresses propranolol sensitivity in infantile hemangioma
Exosomal miR-27a-3p derived from tumor-associated macrophage suppresses propranolol sensitivity in infantile hemangioma
复制标题
源自肿瘤相关巨噬细胞的外泌体 miR-27a-3p 抑制婴儿血管瘤的普萘洛尔敏感性
DOI：
10.1016/j.cellimm.2021.104442
复制
发表时间：
2021-10-08
影响因子：
4.3
通讯作者：
Zheng, Jiawei
中科院分区：
文献类型：
--
作者：
Liu, Chao;Zhao, Zeliang;Zheng, Jiawei
普萘洛尔是治疗婴幼儿血管瘤（IH）的一线药物，而临床观察到普萘洛尔抵抗。肿瘤相关巨噬细胞（TAM）衍生的外泌体可以递送生物分子以促进肿瘤进展。在这里，我们的目的是研究TAM衍生的外泌体miR-27 a-3 p和IH中普萘洛尔敏感性之间的关系。用巨噬细胞集落刺激因子（M-CSF）培养人外周血单核细胞（PBMC）7天，获得未活化的巨噬细胞（Un-Mac），再用IL-4和IL-13诱导M2极化的巨噬细胞。从M2巨噬细胞的条件培养基中分离外泌体，随后进行鉴定。进行细胞共培养和/或转染以探索M2极化的巨噬细胞来源的外泌体（M2-exos）是否可以介导TAM来源的miR-27 a-3 p和血管瘤干细胞（HemSC）之间的串扰。此外，裸鼠皮下移植有用或不用M2-Exos预处理的HemSC，以检查M2-Exos对体内IH的影响。M2极化的巨噬细胞抑制HemSC的普萘洛尔敏感性，如通过增加细胞活力和减少细胞凋亡所示。miR-27 a-3 p在M2极化巨噬细胞和M2-Exos中上调。此外，M2-exos将miR-27 a-3 p从巨噬细胞递送至HemSC，随后降低普萘洛尔敏感性。荧光素酶报告基因和生物素-RNA pulldown实验证明dickkopf相关蛋白2（DKK 2）是miR-27 a-3 p的直接靶点。这些结果表明，M2-exos可将miR-27 a-3 p从巨噬细胞递送至HemSC，通过下调DKK 2表达降低HemSC对普萘洛尔的敏感性，HemSC中的外泌体miR-27 a-3 p和DKK 2可被视为治疗靶点。
Propranolol is the first-line drug for infantile hemangioma (IH) therapy, whereas propranolol resistance is clinically observed. Tumor-associated macrophages (TAMs)-derived exosomes may deliver biological molecules to promote tumor progression. Here, we aimed to investigate the relationship between TAMs-derived exosomal miR-27a-3p and propranolol sensitivity in IH. Human peripheral blood monocytes (PBMCs) were cultured with macrophage colony-stimulating factor (M-CSF) for 7 days to get unactivated macrophages (Un-Mac), which were further treated with IL-4 and IL-13 to induce M2 polarized macrophages. Exosomes were isolated from the conditioned medium of M2 macrophage, followed by identification. Cell co-culture and/or transfection were performed to explore whether M2 polarized macrophage-derived exosomes (M2-exos) could mediate the crosstalk between TAMs-derived miR-27a-3p and hemangioma stem cells (HemSCs). In addition, nude mice were subcutaneously transplanted with HemSCs pretreated with or without M2-Exos to examine the effects of M2-Exos on IH in vivo. M2 polarized macrophages inhibited propranolol sensitivity of HemSCs, as shown by the increased cell viability and decreased apoptosis. miR-27a-3p was upregulated in M2 polarized macrophages and M2-Exos. Moreover, M2-exos delivered miR-27a-3p from macrophages to HemSCs and subsequently reduced propranolol sensitivity. Luciferase reporter and biotin-RNA pulldown assay proved that dickkopf-related protein 2 (DKK2) was the direct target of miR-27a-3p. These results demonstrate that M2-exos could deliver miR-27a-3p from macrophages to HemSCs to reduce the sensitivity of HemSCs to propranolol by down-regulating DKK2 expres-sion, and exosomal miR-27a-3p and DKK2 in HemSCs could be considered as treatment targets.