ASPH-notch Axis guided Exosomal delivery of Prometastatic Secretome renders breast Cancer multi-organ metastasis
ASPH-notch Axis guided Exosomal delivery of Prometastatic Secretome renders breast Cancer multi-organ metastasis
复制标题
ASPH-notch Axis引导的前转移分泌组的外泌体递送使乳腺癌多器官转移
DOI：
10.1186/s12943-019-1077-0
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发表时间：
2019-11-07
期刊：
影响因子：
37.3
通讯作者：
Dong, Xiaoqun
中科院分区：
文献类型：
--
作者：
Lin, Qiushi;Chen, Xuesong;Dong, Xiaoqun
天冬氨酸β羟基酶在正常成人组织中不表达，但在肿瘤发生过程中重新出现，其功能是产生和维持恶性表型所必需的。Exosome使原癌分泌体能够运输和运输，以实现细胞间的远距离通讯。本研究旨在探讨ASPH网络调控特定外切体调控乳腺癌发生发展的分子机制。方法采用慢病毒转染法或CRISPR-Cas9系统建立稳定的ASPH过表达或敲除细胞系。应用免疫印迹、四甲基偶氮唑盐、免疫荧光、荧光素酶报告、免疫共沉淀、2D/3-D侵袭、导管形成、乳房形成、免疫组织化学和新开发的体外转移等方法。结果ASPH通过与Notch受体、配体(Jags)和调节剂(ADAM10/17)的物理相互作用，激活Notch级联反应，为合成/释放促转移外体提供原料(尤其是MMPs/ADAMS)。Exosome协调EMT、2-D/3-D侵袭、茎干、血管生成和转移前的生态位形成。ASPHβ羟基酶的小分子抑制剂(SMI)能特异性/有效地抑制原发部位的基底膜侵袭、血管内/外渗(跨内皮细胞迁移)和远处的定植/生长。在原位和尾静脉注射的小鼠模型中，特定的SMI基本上可以阻断多器官转移。ASPH在正常成人乳腺中是沉默的，在原位恶性肿瘤中表达上调，在浸润性/晚期导管癌中高表达。ASPH的中高表达使乳腺癌具有更多侵袭性的分子亚型(扩增的TNBC或Her2)、早期复发/进展和毁灭性的结果(总体/无病生存率降低)。在乳腺癌患者中，Notch信号元件的表达谱与ASPH的表达呈正相关，证实了ASPH-Notch轴在乳腺肿瘤发生中的作用。结论ASPH-Notch轴引导选择性外体增强多方面的转移。ASPH的促癌/促转移特性对乳腺癌的发生/发展至关重要，揭示了潜在的治疗靶点。
BackgroundAspartate β-hydroxylase (ASPH) is silent in normal adult tissues only to re-emerge during oncogenesis where its function is required for generation and maintenance of malignant phenotypes. Exosomes enable prooncogenic secretome delivering and trafficking for long-distance cell-to-cell communication. This study aims to explore molecular mechanisms underlying how ASPH network regulates designated exosomes to program development and progression of breast cancer.MethodsStable cell lines overexpressing or knocking-out of ASPH were established using lentivirus transfection or CRISPR-CAS9 systems. Western blot, MTT, immunofluorescence, luciferase reporter, co-immunoprecipitation, 2D/3-D invasion, tube formation, mammosphere formation, immunohistochemistry and newly developed in vitro metastasis were applied.ResultsThrough physical interactions with Notch receptors, ligands (JAGs) and regulators (ADAM10/17), ASPH activates Notch cascade to provide raw materials (especially MMPs/ADAMs) for synthesis/release of pro-metastatic exosomes. Exosomes orchestrate EMT, 2-D/3-D invasion, stemness, angiogenesis, and premetastatic niche formation. Small molecule inhibitors (SMIs) of ASPH’s β-hydroxylase specifically/efficiently abrogated in vitro metastasis, which mimics basement membrane invasion at primary site, intravasation/extravasation (transendothelial migration), and colonization/outgrowth at distant sites. Multiple organ-metastases in orthotopic and tail vein injection murine models were substantially blocked by a specific SMI. ASPH is silenced in normal adult breast, upregulated from in situ malignancies to highly expressed in invasive/advanced ductal carcinoma. Moderate-high expression of ASPH confers more aggressive molecular subtypes (TNBC or Her2 amplified), early recurrence/progression and devastating outcome (reduced overall/disease-free survival) of breast cancer. Expression profiling of Notch signaling components positively correlates with ASPH expression in breast cancer patients, confirming that ASPH-Notch axis acts functionally in breast tumorigenesis.ConclusionsASPH-Notch axis guides particularly selective exosomes to potentiate multifaceted metastasis. ASPH’s pro-oncogenic/pro-metastatic properties are essential for breast cancer development/progression, revealing a potential target for therapy.