在临床前小鼠模型中，骨转移性肾细胞癌分泌的细胞外囊泡以时间依赖性方式促进骨髓中的血管生成和内皮间隙形成。
Extracellular vesicles secreted from bone metastatic renal cell carcinoma promote angiogenesis and endothelial gap formation in bone marrow in a time-dependent manner in a preclinical mouse model.
作者信息
Takeda Masashi, Sakamoto Hiromasa, Shibasaki Noboru, Fukui Tomohiro, Magaribuchi Toshihiro, Sumiyoshi Takayuki, Utsunomiya Noriaki, Sawada Atsuro, Goto Takayuki, Kobayashi Takashi, Ueda Koji, Yamasaki Toshinari, Ogawa Osamu, Akamatsu Shusuke
机构信息
Department of Urology, Kyoto University Graduate School of Medicine, Kyoto, Japan.
Project for Personalized Cancer Medicine, Cancer Precision Medicine Center, Japanese Foundation for Cancer Research, Tokyo, Japan.
出版信息
Front Oncol. 2023 Mar 23;13:1139049. doi: 10.3389/fonc.2023.1139049. eCollection 2023.
INTRODUCTION
Bone is a major metastatic site of renal cell carcinoma (RCC). Recently, it is well recognized that bone metastatic tumor cells remodel bone marrow vasculature. However, the precise mechanism underlying cell-cell communication between bone metastatic RCC and the cells in bone marrow remains unknown. Extracellular vesicles (EVs) reportedly play crucial roles in intercellular communication between metastatic tumor cells and bone marrow. Therefore, we conducted the current study to clarify the histological alteration in vascular endothelium in bone marrow induced by EVs secreted from bone metastatic RCC cells as well as association between angiogenesis in bone marrow and bone metastasis formation.
MATERIALS AND METHODS
We established a bone metastatic RCC cell line (786-O BM) by in vivo selection and observed phenotypic changes in tissues when EVs were intravenously injected into immunodeficient mice. Proteomic analysis was performed to identify the protein cargo of EVs that could contribute to histological changes in bone. Tissue exudative EVs (Te-EVs) from cancer tissues of patients with bone metastatic RCC (BM-EV) and those with locally advanced disease (LA-EV) were compared for in vitro function and protein cargo.
RESULTS
Treatment of mice with EVs from 786-O BM promoted angiogenesis in the bone marrow in a time-dependent manner and increased the gaps of capillary endothelium. 786-O BM EVs also promoted tube formation . Proteomic analysis of EVs identified aminopeptidase N (APN) as a candidate protein that enhances angiogenesis. APN knockdown in 786-O BM resulted in reduced angiogenesis in vitro and in vivo. When parental 786-O cells were intracardially injected 12 weeks after treatment with786-O BM EVs, more bone metastasis developed compared to those treated with EVs from parental 786-O cells. In patient samples, BM-EVs contained higher APN compared to LA-EV. In addition, BM-EVs promoted tube formation in vitro compared to LA-EVs.
CONCLUSION
EVs from bone metastatic RCC promote angiogenesis and gap formation in capillary endothelium in bone marrow in a time-dependent manner.
引言
骨是肾细胞癌（RCC）的主要转移部位。最近，人们已经充分认识到骨转移肿瘤细胞会重塑骨髓血管系统。然而，骨转移性RCC与骨髓细胞之间细胞间通讯的确切机制仍不清楚。据报道，细胞外囊泡（EVs）在转移性肿瘤细胞与骨髓之间的细胞间通讯中起关键作用。因此，我们进行了本研究，以阐明骨转移性RCC细胞分泌的EVs诱导的骨髓血管内皮组织学改变，以及骨髓血管生成与骨转移形成之间的关联。
材料与方法
我们通过体内筛选建立了骨转移性RCC细胞系（786-O BM），并观察了将EVs静脉注射到免疫缺陷小鼠体内时组织的表型变化。进行蛋白质组学分析以鉴定可能导致骨组织学变化的EVs蛋白质成分。比较了骨转移性RCC患者（BM-EV）和局部晚期疾病患者（LA-EV）癌组织的组织渗出性EVs（Te-EVs）的体外功能和蛋白质成分。
结果
用786-O BM的EVs处理小鼠以时间依赖性方式促进了骨髓血管生成，并增加了毛细血管内皮的间隙。786-O BM EVs还促进了管形成。对EVs的蛋白质组学分析确定氨肽酶N（APN）是一种增强血管生成的候选蛋白质。786-O BM中的APN敲低导致体外和体内血管生成减少。在用786-O BM EVs处理12周后经心内注射亲代786-O细胞时相比于用亲代786-O细胞的EVs处理的小鼠，发生了更多的骨转移。在患者样本中，与LA-EV相比，BM-EV含有更高水平的APN。此外，与LA-EV相比，BM-EV在体外促进了管形成。
结论
骨转移性RCC的EVs以时间依赖性方式促进骨髓毛细血管内皮的血管生成和间隙形成。
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