SPECIAL FOCUS: EMERGING IMPACT OF EXTRACELLULAR VESICLES ON TISSUE ENGINEERING AND REGENERATION* A Systematic Evaluation of Factors Affecting Extracellular Vesicle Uptake by Breast Cancer Cells
SPECIAL FOCUS: EMERGING IMPACT OF EXTRACELLULAR VESICLES ON TISSUE ENGINEERING AND REGENERATION* A Systematic Evaluation of Factors Affecting Extracellular Vesicle Uptake by Breast Cancer Cells
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发表时间：
2017
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通讯作者：
Devin M. Stranford;Michelle E. Hung;Emma S Gargus;R. Shah;J. Leonard
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作者：
Devin M. Stranford;Michelle E. Hung;Emma S Gargus;R. Shah;J. Leonard
细胞外囊泡是由细胞分泌的纳米级颗粒，通过在细胞间转移生物分子来介导细胞间通讯。利用这种机制进行治疗性生物分子递送代表了再生医学和其他临床应用的一个有前途的前沿。实现这一目标的一个挑战是，到目前为止，我们对哪些因素影响受体细胞对EV的吸收的理解仍然不完整。在这项研究中，我们系统地研究了乳腺癌细胞的背景下，这是一个最充分研究的细胞类型的EV交付，因此包括一个简单的模型系统，这项调查的交付问题。通过在EV表面上展示各种靶向肽，我们观察到尽管在EV上展示GE 11适度增加了MCF-7细胞的摄取，但神经肽Y（NPY）展示对相同细胞的摄取没有影响。相比之下，神经降压素（NTS）和尿激酶纤溶酶原激活剂（uPA）显示减少的EV摄取的MDA-MB-231细胞。有趣的是，EV吸收率并不取决于EV的来源;与HEK 293 FT衍生的EV相比，乳腺癌细胞对乳腺癌衍生的EV的吸收没有增加。此外，EV摄取大大增强了交付的聚凝胺和spinoculation的存在下，表明最大EV摄取率是远远大于在细胞培养的基础条件下观察到的。通过研究细胞的环境如何提供影响EV摄取的线索，我们还观察到，与在硬组织培养聚苯乙烯上培养相比，在软基质上培养细胞显著增强了EV摄取。这些观察结果中的每一个都提供了对影响乳腺癌细胞EV摄取的因素的见解，同时也为系统地评估和可能增强各种细胞类型的EV摄取提供了比较基础。
Extracellular vesicles (EVs) are nanometer-scale particles that are secreted by cells and mediate intercellular communication by transferring biomolecules between cells. Harnessing this mechanism for therapeutic biomolecule delivery represents a promising frontier for regenerative medicine and other clinical applications. One challenge to realizing this goal is that to date, our understanding of which factors affect EV uptake by recipient cells remains incomplete. In this study, we systematically investigated such delivery questions in the context of breast cancer cells, which are one of the most well-studied cell types with respect to EV delivery and therefore comprise a facile model system for this investigation. By displaying various targeting peptides on the EV surface, we observed that although displaying GE11 on EVs modestly increased uptake by MCF-7 cells, neuropeptide Y (NPY) display had no effect on uptake by the same cells. In contrast, neurotensin (NTS) and urokinase plasminogen activator (uPA) display reduced EV uptake by MDA-MB-231 cells. Interestingly, EV uptake rate did not depend on the source of the EVs; breast cancer cells demonstrated no increase in uptake on administration of breast cancer-derived EVs in comparison to HEK293FT-derived EVs. Moreover, EV uptake was greatly enhanced by delivery in the presence of polybrene and spinoculation, suggesting that maximal EV uptake rates are much greater than those observed under basal conditions in cell culture. By investigating how the cell’s environment might provide cues that impact EV uptake, we also observed that culturing cells on soft matrices significantly enhanced EV uptake, compared to culturing on stiff tissue culture polystyrene. Each of these observations provides insights into the factors impacting EV uptake by breast cancer cells, while also providing a basis of comparison for systematically evaluating and perhaps enhancing EV uptake by various cell types.