Center on Probes for Molecular Mechanotechnology

分子机械技术探针中心

• 批准号: 10629919
• 负责人: Khalid S Salaita
• 金额: $ 146.8万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-10 至 2028-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10629919
• 关键词: Abnormal Cell; Acceleration; Adaptive Immune System; Address; Adoption; Affect; Apple; Architecture; Automobile Driving; Binding; Biomedical Technology; Blood Platelets; Blood Transfusion; Cardiopulmonary Bypass; Cell Surface Receptors; Cell physiology; Cells; Cellular biology; Clinical Research; Coagulation Process; Collaborations; Communities; Custom; DNA; DNA Probes; DNA Structure; DNA amplification; Data Analyses; Diagnosis; Disease; Dissociation; Educational workshop; Embryo; Ensure; Enzyme-Linked Immunosorbent Assay; Event; Extramural Activities; Fees; Flow Cytometry; Fluorescence Polarization; Focal Adhesions; Frequencies; Functional disorder; Geography; Goals; Growth; Heterogeneity; Image; Immune response; Industry; Internet; Invaded; Knowledge; Life; Mainstreaming; Malignant neoplasm of lung; Maps; Measures; Mechanics; Mediating; Methods; Microscope; Molecular; Molecular Probes; National Institute of General Medical Sciences; Neoplasm Metastasis; Normal Cell; Pathway interactions; Patients; Peer Review; Preparation; Process; Proteins; Proteomics; Protocols documentation; Publications; Publishing; Reader; Reagent; Research Personnel; Resolution; Role; Seminal; Series; Services; Signal Transduction; Stroke; Structure; Surface; System; T-Cell Immunologic Specificity; T-Cell Receptor; T-Lymphocyte; Techniques; Technology; Testing; Time; Traction; Training; Weight; Work; adhesion receptor; cancer cell; cell growth; commercialization; community engagement; design; force sensor; high throughput analysis; improved; interest; mechanical force; mechanical signal; mechanotransduction; nanoscale; new technology; prototype; public health relevance; receptor; recruit; response; symposium; technology development; tool; virtual

Project summary The proposed Center on Probes for Molecular Mechanotechnology (CPMM) will work to develop and optimize technologies to enable the study of mechanobiology and mechanotransduction pathways in living cells. The CPMM includes three highly synergistic Technology Development Projects (TDPs) that will be led by Alexa Mattheyses, Khalid Salaita, and Yonggang Ke who have a strong track record of jointly publishing and working together to developed tension probe technologies. In TDP#1: High resolution probes for mechanobiology, we will create “indestructible” probes that can push the limits of spatial and orientation resolution for the DNA tension probe technology. Tension-PAINT imaging will be refined to achieve realtime 20 nm spatial mapping of forces and to combine this with immunostaining to map the proteins that assemble within proximity to mechanically active receptors. Force orientation will be mapped using fluorescence polarization methods with turn-key commercial microscopes. In TDP#2: Probes for mechanical tagging, we will develop methods of force-induced tagging. The central design feature is a DNA probe that mediates a binding event or dissociation event at threshold levels of force. Cells are tagged based on the magnitude and frequency of mechanical events generated by a cell surface receptors. This TDP will lead to high-throughput analysis of cells using flow cytometry and will also allow for proteomic analysis to open the door to “mechanomics”. Under TDP#3: Amplified force sensors, the central technology here is responsive DNA structures that amplify mechanical inputs. The CPMM has nine associated inaugural Driving Biomedical Projects (DBPs) led by a team of geographically diverse collaborators. DBPs #1-#4 are focused on mechanobiology of T cells and use CPMM tools to test the mechanosensor function of the T cell receptor (TCR) and the adhesion receptor LFA-1. DBP#5 focuses on the heterogeneity in cancer cells. DBP#6 and #7 target the mechanosensor responses of platelets. Finally, DBP#8 and #9 address fundamental questions of the role of mechanics in focal adhesions. Our prototype TDP technologies provide methods to measure molecular forces with the same ease and simplicity as that of immunostaining, flow cytometry, PCR and ELISA. But unlike these mainstream techniques, mechano-imaging, mechano-PCR, mechano-flow, and mechano-ELISA are not commercialized. Hence, the reagents and surface preparation protocols and data analysis routines have to be custom prepared by the end user. This can be challenging to the non-expert and is not routine. Therefore, the CPMM will integrate a strong Community Engagement (CE) component. CE activities will focus on hands-on training workshops, publication of methods articles, virtual seminar series, industry engagement, a strong web presence and engagement with three key mechanobiology conferences that will help accelerate adoption of the tension probe technology. These CE activities will ultimately lead to commercialization which will enable wide spread dissemination across the various cell biology communities.

项目总结 拟建的分子力学技术探针中心(CPMM)将致力于发展和优化 能够研究活细胞中的机械生物学和机械转导途径的技术。这个 CPMM包括三个高度协同的技术开发项目(TDP)，这些项目将由Alexa领导 马修斯、哈立德·萨莱塔和柯永刚，他们有很强的联合出版和工作记录 共同开发了张力探头技术。在TDP#1：机械生物学的高分辨率探测器中，我们 将创造“坚不可摧”的探测器，可以突破dna张力的空间和方位分辨率的极限。 探头技术。张力-油漆成像将得到改进，以实现力的实时20 nm空间映射 并将其与免疫染色相结合，以绘制机械地在邻近区域组装的蛋白质图 活跃的受体。将使用带有交钥匙的荧光偏振法绘制部队方向图 商用显微镜。在TDP#2：用于机械标记的探针中，我们将发展力诱导的方法 标记。中心设计特征是一个DNA探针，它在 武力的门槛水平。单元格根据机械事件的大小和频率进行标记 由细胞表面受体产生。这种TDP将导致使用流式细胞术对细胞进行高通量分析 并将允许蛋白质组分析打开“机制组学”的大门。在TDP#3下：扩大力量 传感器，这里的核心技术是响应性DNA结构，它放大机械输入。CPMM 有九个相关的启动生物医学项目(DBP)，由一个地理位置不同的团队领导 合作者。DBP#1-#4专注于T细胞的机械生物学，并使用CPMM工具测试 T细胞受体(TCR)和黏附受体LFA-1的机械传感器功能。DBP#5重点关注 癌细胞的异质性。DBP#6和#7针对的是血小板的机械传感器反应。最后，DBP#8 和#9解决了力学在局部粘连中所起作用的基本问题。我们的TDP原型 技术提供了测量分子力的方法，其简单易行 免疫组织化学染色、流式细胞术、聚合酶链式反应和酶联免疫吸附试验。但与这些主流技术不同，机械成像， 机械式聚合酶链式反应、机械式流动免疫吸附试验和机械式酶联免疫吸附试验均未商业化。因此，试剂和表面 准备方案和数据分析例程必须由最终用户定制准备。这可以是 挑战非专家，这不是例行公事。因此，CPMM将整合一个强大的社区 接洽(CE)组件。CE的活动将侧重于动手培训讲习班、出版方法 文章、虚拟研讨会系列、行业参与度、强大的网络表现和参与度 机械生物学会议将有助于加快张力探头技术的采用。这些行政长官 活动最终将导致商业化，这将使各种不同的 细胞生物学群落。