单细胞水平实时可视研究超声联合微泡介导质膜穿孔后修复的机制

项目名称： 单细胞水平实时可视研究超声联合微泡介导质膜穿孔后修复的机制

项目编号： No.81471667

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 医药、卫生

项目作者： 秦鹏

作者单位： 上海交通大学

项目金额： 73万元

中文摘要： 低强度超声联合微泡能够介导质膜穿孔，递送大分子药物进入活细胞，其无创安全、低成本、双靶向性的特点彰显了这一物理技术用于药物传输的优势。质膜穿孔后的修复过程和细胞命运息息相关，对药物传输效率及生物安全性至关重要。本研究以多种细胞系为模型，创建单细胞水平上实时可视平台，从时空域揭示超声联合微泡致膜穿孔后修复这一短暂快速、微观复杂的生物过程。基于此，采取多种生物学实验技术进行以下研究：（1）稳态空化致膜穿孔和细胞骨架响应、机械敏感蛋白及胞吞的时空域关系，确定参与质膜修复的机械传导通路；（2）确定胞内钙离子暂流、溶酶体胞吐及细胞骨架动态行为在瞬态空化致膜穿孔后修复的作用机理；（3）确定不同空化剂量致膜穿孔后细胞的命运。通过该项研究，期望深入揭示超声联合微泡介导质膜穿孔后修复的生物学机理，并提出促进质膜修复，改进药物传输效率，提高生物安全性的策略，为其走向临床应用奠定可靠、充分的理论基础。

中文关键词： 超声；微泡；声致穿孔；质膜修复；药物传输

英文摘要： Sonoporation refers to the temporal cell membrane opening caused by the interaction between the low intensity ultrasound pulses and microbubbles, resulting in facilitation of the transport of membrane impermeable macromolecular into living cells. The merits of non-invasion, dual-targeting to malignant tissues, safety and low cost make this physical delivery approach promising for clinical drug delivery.The resealing of the transient membrane poration induced by ultrasound and microbubbles is closely correlated with the fate of targeted cells, and therefore is important for the efficiency and biosafety of drug delivery. Based on the establishment of a real-time visualization platform at the single-cell level, diverse tumor cell lines were adopted to reveal the kinetics and microscopic process of membrane repair after acoustic cavitation in a spatial and temporal manner.Based on the above system, various experimental technologies will be utilized to study the following scientific problems. (1) The spatiotemporal relation in the cytoskeleton response, mechanosensitive channels and exocytosis after stable cavitation induced by sonoporation will be analyzed; and specific mechanotransduction pathways responsive for the membrane repair after stable cavitation will be studied. (2) The role of the intracellular calcium transient, lysosome exdocytosis and cytoskeleton dynamic behaviors in the membrane resealing after inertial cavitation induced membrane damage will be revealed. (3) The cell fate in the long period after treatment with different acoustic cavitation doses will be determined.These studies will provide us a detailed understanding of the repair mechanisms after the membrane perforations induced by ultrasound and microbubbles, and will propose the strategies for promoting the membrane resealing and improving delivery efficiency and biosafety of ultrasound induced drug delivery. These findings will also establish a solid theoretical foundation for clinical drug delivery by the ultrasound with combination of microbubbles.

英文关键词： Ultrasound;Microbubble;Sonoporation;Membrane Resealing;Drug Delivery