Endocytic pathway inhibition attenuates extracellular vesicle-induced reduction of chemosensitivity to bortezomib in multiple myeloma cells.
Endocytic pathway inhibition attenuates extracellular vesicle-induced reduction of chemosensitivity to bortezomib in multiple myeloma cells.
复制标题
内吞途径抑制减弱细胞外囊泡诱导的多发性骨髓瘤细胞对硼替佐米化疗敏感性的降低
作者：
Tu C;Du Z;Zhang H;Feng Y;Qi Y;Zheng Y;Liu J;Wang J
来源于骨髓基质细胞（BMSCs）的细胞外囊泡（EV），包括外泌体和微囊泡，已被证明是多发性骨髓瘤（MM）进展和耐药的关键因素。EV摄取涉及多种机制，这在很大程度上取决于囊泡来源和受体细胞类型。本研究的目的是确定骨髓基质细胞来源的小EV（sEV）的MM细胞的摄取机制，并评估针对这一过程的抗MM效果。方法：应用透射电镜、纳米颗粒示踪分析和蛋白质印迹法对人骨髓基质细胞来源的sEV进行鉴定。流式细胞仪分析化学抑制剂和shRNA介导的内吞相关基因敲低对sEV摄取和细胞凋亡的影响。在体外和异种移植MM小鼠模型中评价了阻断sEV摄取的抗MM作用。结果如下：来源于BMSC的sEV以时间和剂量依赖性的方式被MM细胞吸收，随后促进MM细胞周期并降低其对硼替佐米的化学敏感性。靶向硫酸肝素蛋白聚糖、肌动蛋白、酪氨酸激酶、发动蛋白-2、钠/质子交换剂或磷酸肌醇3-激酶的化学内吞抑制剂显著降低了骨髓基质细胞衍生的sEV的MM细胞内化。此外，在MM细胞中，shRNA介导的内吞相关蛋白（包括小窝蛋白-1、漂浮蛋白-1、网格蛋白重链和发动蛋白-2）的敲低抑制sEV摄取。此外，靶向发动蛋白-2的内吞作用抑制剂优先抑制原代MM细胞离体摄取sEV，并增强硼替佐米在体外和小鼠模型中的抗MM作用。结论：网格蛋白和小窝蛋白依赖的内吞作用和巨胞饮作用是sEV介导的BMSC和MM细胞之间的通信的主要途径，并且抑制内吞作用减弱sEV诱导的对硼替佐米的化学敏感性降低，从而增强其抗MM特性。
Extracellular vesicles (EVs), including exosomes and microvesicles, derived from bone marrow stromal cells (BMSCs) have been demonstrated as key factors in the progression and drug resistance of multiple myeloma (MM). EV uptake involves a variety of mechanisms which largely depend on the vesicle origin and recipient cell type. The aim of the present study was to identify the mechanisms involved in the uptake of BMSC-derived small EVs (sEVs) by MM cells, and to evaluate the anti-MM effect of targeting this process. Methods: Human BMSC-derived sEVs were identified by transmission electron microscopy, nanoparticle tracking analysis, and western blot. The effects of chemical inhibitors and shRNA-mediated knockdown of endocytosis-associated genes on sEV uptake and cell apoptosis were analyzed by flow cytometry. The anti-MM effect of blocking sEV uptake was evaluated in vitro and in a xenograft MM mouse model. Results: sEVs derived from BMSC were taken up by MM cells in a time- and dose-dependent manner, and subsequently promoted MM cell cycling and reduced their chemosensitivity to bortezomib. Chemical endocytosis inhibitors targeting heparin sulphate proteoglycans, actin, tyrosine kinase, dynamin-2, sodium/proton exchangers, or phosphoinositide 3-kinases significantly reduced MM cell internalization of BMSC-derived sEVs. Moreover, shRNA-mediated knockdown of endocytosis-associated proteins, including caveolin-1, flotillin-1, clathrin heavy chain, and dynamin-2 in MM cells suppressed sEV uptake. Furthermore, an endocytosis inhibitor targeting dynamin-2 preferentially suppressed the uptake of sEV by primary MM cells ex vivo and enhanced the anti-MM effects of bortezomib in vitro and in a mouse model. Conclusion: Clathrin- and caveolin-dependent endocytosis and macropinocytosis are the predominant routes of sEV-mediated communication between BMSCs and MM cells, and inhibiting endocytosis attenuates sEV-induced reduction of chemosensitivity to bortezomib, and thus enhances its anti-MM properties.
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影响因子：
7.4
作者：
Marleau AM;Chen CS;Joyce JA;Tullis RH
通讯作者：
Tullis RH
影响因子：
11.8
作者：
Macia, Eric;Ehrlich, Marcelo;Kirchhausen, Tomas
通讯作者：
Kirchhausen, Tomas
影响因子：
20.3
作者：
Bender, Markus;Giannini, Silvia;Falet, Herve
通讯作者：
Falet, Herve
影响因子：
37.3
作者：
Iaccino E;Mimmi S;Dattilo V;Marino F;Candeloro P;Di Loria A;Marimpietri D;Pisano A;Albano F;Vecchio E;Ceglia S;Golino G;Lupia A;Fiume G;Quinto I;Scala G
通讯作者：
Scala G
影响因子：
4
作者：
Fitzner, Dirk;Schnaars, Mareike;Simons, Mikael
通讯作者：
Simons, Mikael