Development of a mechanosensitive synthetic cell for mediating intercellular communication.

项目摘要 我们设计能够与活细胞交流的合成细胞系统的能力仍然有限。的 长期目标是设计具有越来越复杂的仿生功能的细胞样系统， 作为细胞替代或增强天然细胞的功能。本建议的目的是制定一个 机械敏感的合成细胞，可以响应剪切应力的增加，这是最普遍的， 心血管系统，并分泌生物活性分子影响活细胞。我们体内的细胞 不断感知和响应微环境刺激，包括被动和主动的物理刺激， 例如细胞外基质刚性、粘附配体密度、张力、压缩和流体剪切流。的 这一提议的基本原理是，完成将导致一种新的仿生细胞样系统， 新型剪切应力响应“材料”，可以与天然活细胞接触。大多数 工程生物材料响应于生物化学环境的差异（例如氧化还原的差异， pH和酶组成）。相比之下， 在利用刺激反应行为的力量方面相对较少的努力。合成细胞的想法 通过天然血小板的结合能力和对升高的剪切应力的反应以及分泌颗粒内容物， 绑定到一个表面。本文的工作包括三个具体目标：1）表征剪切应力 机械感测囊泡的响应，2）在合成细胞中将机械感测与胞吐作用耦合，3） 体外检测切应力激活的合成细胞与内皮细胞的细胞间通讯。我们 我将使用一种创新的方法来实现这些目标，即重新利用机械敏感通道进行剪切 应力传感和使用基于肽的膜融合。我们的实验室是第一个展示 机械感应合成细胞，我们在自下而上的合成生物学方面拥有重要的专业知识。的 这项研究意义重大，因为它将是第一个开发出来的用于交流的合成细胞系统。 与哺乳动物细胞的钙触发分泌。这项工作将制定基本战略， 将机械感测与合成细胞中的生化反应耦合。这将为其他国家开辟新的途径。 对开发更复杂的细胞样系统感兴趣的研究人员。其结果将对 因为它将支持机械敏感通道可以感知 侧膜张力由于剪应力而长期存在， 工程合成细胞与其他传感能力。

Project Summary Our abilities to engineer synthetic cell systems that can communicate with living cells remain limited. The long-term goal is to engineer cell-like systems with increasingly complex biomimetic functions that can serve as cell replacement or augment functions of natural cells. The objective of this proposal is to develop a mechanosensitive synthetic cell that can respond to an increase in shear stress, which is most prevalent in the cardiovascular system, and secrete bioactive molecules to effect living cells. Cells in our bodies constantly sense and respond to microenvironmental stimuli, including passive and active physical stimuli, such as extracellular matrix rigidity, adhesive ligand density, tension, compression, and fluid shear flow. The rationale underlying this proposal is that completion will result in a novel biomimetic cell-like system as a novel shear stress-responsive ‘material’ that can interface with natural living cells. The majority of engineered biomaterials respond to differences in the biochemical environment (e.g. differences in redox, pH, and enzyme composition) between normal and diseased tissues. By comparison, there has been relatively less effort in exploiting forces for stimulus-responsive behaviors. The synthetic cell idea is inspired by natural platelets’ ability to bind and respond to elevated shear stress and secrete granule contents when bound to a surface. The proposed work consists of three specific aims: 1) Characterize shear stress response of mechanosensing vesicles, 2) Couple mechanosensing with exocytosis in synthetic cells, 3) Test intercellular communication of shear stress-activated synthetic cells with endothelial cells in vitro. We will pursue these aims using an innovative approach of repurposing mechanosensitive channel for shear stress sensing and using peptide-based membrane fusion. Our lab was the first group to demonstrate mechanosensing synthetic cells and we have significant expertise in bottom-up synthetic biology. The proposed research is significant, because it will be the first synthetic cell system developed to communicate with mammalian cells using calcium-triggered secretion. The work will develop fundamental strategies for coupling mechanosensing to a biochemical response in synthetic cells. This will open new avenue for other researchers interested in developing more complex cell-like systems. The results will have an important positive impact immediately because it will support the idea that mechanosensitive channels can sense lateral membrane tension due to shear stress and long-term because they lay the groundwork of engineering synthetic cells with other sensing abilities.