Collaborative Research: A Nanostructured Model of the Apoptotic Cell Surface

合作研究：凋亡细胞表面的纳米结构模型

• 批准号: 1033215
• 负责人: Michelle Knowles
• 金额: $ 18.44万
• 依托单位: University of Denver
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-10-01 至 2015-09-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1033215&HistoricalAwards=false
• 关键词: Collaborative; Research; Nanostructured; Model; Apoptotic

AbstractCollaborative Research: A Nanostructured Model of the Apoptotic Cell SurfaceSensors that mimic cellular membranes have contributed to understanding of protein-membrane interactions. However, the membranes of an apoptotic cells differ in lipid composition and structure, containing oxidized lipids that cause nanoscale and microscale protrusions. Membrane shape is a potential site for protein recognition and a critical way in which the body identifies damaged cells for removal. By mimicking the shapes of apoptotic membranes, sensors will be created that more accurately reflect apoptotic cells. Nanostructured membranes will be created using supported bilayer techniques. These nanostructured models of the apoptotic cell surface will allow for separate control of both membrane curvature and lipid composition. Binding of protein to these sensors will be monitored using a surface sensitive, fluorescence microscopy technique. Sensors designed using this approach will be useful for understanding how proteins interact with curved membrane surfaces. The first goal of this research is to construct supported bilayer membranes that allow for control of lipid composition as well as nanoscale surface structure. Silica nanoparticles embedded with fluorophores will be prepared and coated with a series of oxidized and non-oxidized lipids. These nanoparticles will be attached to glass surfaces and a lipid bilayer will be formed over them. The second goal of this research is to demonstrate the ability of these nanostructured membrane sensors to measure protein binding to curved surfaces. The effects of curvature on the binding of proteins involved with recognizing apoptotic cells for removal will be measured using total internal reflection fluorescence microscopy. Binding of C-reactive protein to the membranes will be visualized and quantitatively measured to determine what features of an apoptotic membrane are critical for protein recognition. This information will guide the future design of sensors that detect how the body responds to apoptotic cells.Intellectual Merits: It is unknown whether lipids, membrane curvature, or the combination determines protein binding to apoptotic cells. This work will answer that question and improve understanding of apoptotic cell recognition. In turn, this will allow for the design of sensors that recognize physiological responses to apoptotic cells that could be critical in diagnosing cardiovascular disease. The approaches developed here for engineering mimics of apoptotic cell membranes will become the foundation for sensors based on optical, electronic, or mass based signal transduction.Broader impacts: The proposed research will impact teaching, training, and outreach. A postdoctoral scientist and several students will be trained in emerging techniques at the interface of nanoscience, biology, chemistry, and spectroscopy. Additionally, the educational impact of this project will extend beyond the two campuses and will reach out to involve underrepresented minorities. Students from the Strides Toward Encouraging Professions in Science program at the Community College of Aurora, Community College of Denver and Metropolitan State College of Denver will be involved in the proposed research. A second method of outreach will be to include middle school science teachers through the Rocky Mountain-Middle School Math and Science Partnership.

摘要调查研究：模拟细胞膜的凋亡细胞表面体的纳米结构模型有助于理解蛋白质 - 膜相互作用。 然而，凋亡细胞的膜在脂质组成和结构上有所不同，其中包含引起纳米级和微观突起的氧化脂质。 膜形状是蛋白质识别的潜在部位，也是人体识别受损细胞去除的关键方法。 通过模仿凋亡膜的形状，将创建更准确地反映凋亡细胞的传感器。 纳米结构的膜将使用支持的双层技术创建。这些凋亡细胞表面的纳米结构模型将可以单独控制膜曲率和脂质组成。将使用表面敏感的荧光显微镜技术来监测蛋白质与这些传感器的结合。使用这种方法设计的传感器将有助于了解蛋白质如何与弯曲膜表面相互作用。这项研究的第一个目标是构建支持的双层膜，以控制脂质成分以及纳米级的表面结构。 嵌入荧光团的二氧化硅纳米颗粒将与一系列氧化和非氧化脂质涂在一起。这些纳米颗粒将连接到玻璃表面，并在其上形成脂质双层。 这项研究的第二个目标是证明这些纳米结构膜传感器测量蛋白质与弯曲表面的结合的能力。 曲率对识别凋亡细胞去除的蛋白质的结合的影响将使用总内反射荧光显微镜进行测量。 C反应蛋白与膜的结合将被观察并进行定量测量，以确定凋亡膜的哪些特征对于蛋白质识别至关重要。 该信息将指导传感器的未来设计，这些传感器检测人体如何对凋亡细胞的反应。智能优点：尚不清楚脂质，膜曲率或组合是否决定蛋白质与凋亡细胞的结合。 这项工作将回答这个问题，并提高对凋亡细胞识别的理解。 反过来，这将允许设计传感器，以识别对凋亡细胞的生理反应，这对于诊断心血管疾病至关重要。 此处开发的用于工程模仿凋亡细胞膜的方法将成为基于光学，电子或基于质量的信号转导的传感器的基础。BRODER的影响：拟议的研究将影响教学，培训和外展。一位博士后科学家和几位学生将接受纳米科学，生物学，化学和光谱学界面的新兴技术培训。 此外，该项目的教育影响将超出两个校园，并将涉及代表性不足的少数民族。 从大步前进到鼓励奥罗拉社区学院，丹佛社区学院和丹佛大都会州立学院的科学课程专业的学生将参与拟议的研究。 宣传的第二种方法是通过落基山中学学校数学和科学伙伴关系来包括中学科学老师。