Development of a mechanosensitive synthetic cell for mediating intercellular communication.

开发用于介导细胞间通讯的机械敏感合成细胞。

• 批准号: 10031135
• 负责人: Allen Po-Chih Liu
• 金额: $ 27.05万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-02 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10031135
• 关键词: Adhesions; Adhesives; Apoptosis; Behavior; Binding; Biochemical; Biocompatible Materials; Biological Markers; Biomimetics; Blood Platelets; Blood Vessels; Calcium; Calpain; Cardiovascular system; Cell Death; Cell model; Cells; Communication; Complex; Coupling; Cytoplasmic Granules; Data; Development; Disease; Encapsulated; Endothelial Cells; Engineering; Environment; Enzymes; Epidermal Growth Factor Receptor; Exocytosis; Extracellular Matrix; Fostering; Future; Goals; Growth; Health; Human body; Hybrids; Immunotherapy; In Vitro; Lateral; Ligands; Lipid Bilayers; Liquid substance; Location; Lytic; Mammalian Cell; Measures; Mechanics; Mediating; Membrane; Membrane Fusion; Mission; Modeling; Monitor; Morphology; Oxidation-Reduction; Pathology; Patients; Peptides; Physiological; Polymers; Probability; Process; Property; Public Health; Research; Research Personnel; Signal Transduction; Signal Transduction Pathway; Specificity; Stimulus; Surface; System; T-Lymphocyte; Techniques; Technology; Testing; Therapeutic; Tissues; United States National Institutes of Health; Variant; Vascular Endothelial Growth Factors; Vesicle; Work; base; biological adaptation to stress; cancer cell; cell killing; cell type; chimeric antigen receptor; density; exosome; experience; hemodynamics; improved; innovation; intercellular communication; interest; mechanical force; mechanotransduction; membrane reconstitution; novel; prevent; receptor binding; reconstitution; response; shear stress; stem cells; success; synthetic biology; tool; tumor

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.

项目概要 我们设计能够与活细胞通信的合成细胞系统的能力仍然有限。这 长期目标是设计具有日益复杂的仿生功能的类细胞系统，以服务于 作为细胞替代或增强自然细胞的功能。该提案的目标是开发一个 机械敏感的合成细胞，可以响应剪切应力的增加，这在 心血管系统，并分泌生物活性分子来影响活细胞。我们体内的细胞 不断感知并响应微环境刺激，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。这 该提案的基本原理是，完成后将产生一种新型仿生细胞样系统，作为 新型剪切应力响应“材料”，可以与天然活细胞相互作用。大部分 工程生物材料对生化环境的差异做出反应（例如氧化还原的差异， 正常组织和患病组织之间的 pH 值和酶组成）。相比之下，已有 利用力量进行刺激反应行为的努力相对较少。合成细胞的想法受到启发 通过天然血小板结合和响应升高的剪切应力并分泌颗粒内容物的能力 绑定到表面。拟议的工作包括三个具体目标：1）表征剪切应力 机械传感囊泡的响应，2) 将机械传感与合成细胞中的胞吐作用结合起来，3) 体外测试剪切应力激活的合成细胞与内皮细胞的细胞间通讯。我们 将使用一种创新方法来实现这些目标，重新利用剪切力的机械敏感通道 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的小组 机械传感合成细胞，我们在自下而上的合成生物学方面拥有丰富的专业知识。这 拟议的研究意义重大，因为它将是第一个开发用于通信的合成细胞系统 哺乳动物细胞利用钙触发分泌。这项工作将制定基本战略 将机械传感与合成细胞中的生化反应耦合。这将为其他领域开辟新途径 对开发更复杂的类细胞系统感兴趣的研究人员。结果将具有重要意义 立即产生积极影响，因为它将支持机械敏感通道可以感知的想法 由于剪切应力和长期的侧向膜张力，因为它们奠定了基础 工程合成细胞具有其他传感能力。