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.

项目摘要 提议的分子机械技术探针中心（CPMM）将有效地开发和优化 技术能够研究活细胞中的机械生物学和机械转导途径。这 CPMM包括三个由Alexa领导的高度协同技术开发项目（TDP） Mattheyses，Khalid Salaita和Yonggang KE，在共同出版和工作方面有着良好的记录 在TDP＃1中：机械生物学的高分辨率问题，我们 将创建“可索引”的问题，可以推动DNA张力的空间和方向分辨率的限制 探针技术。张力涂料成像将被完善，以实现实时20 nm的力量映射力 并将其与免疫染色结合起来，以绘制在机械近端组装的蛋白质 活跃的接收器。力取向将使用荧光偏振法映射 商业显微镜。在TDP＃2：机械标记的探针中，我们将开发力诱导的方法 标记。中心设计特征是DNA探针，该探针介导了结合事件或解离事件 阈值的力水平。根据机械事件的幅度和频率标记细胞 由细胞表面受体产生。该TDP将通过流式细胞仪对细胞进行高通量分析 并且还将允许蛋白质组学分析打开“机械学”的门。在TDP＃3下：放大力 传感器，这里的中央技术是放大机械输入的响应式DNA结构。 CPMM 由一个由地理上的潜水员团队领导的九个相关的就职生物医学项目（DBP） 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）和粘合受体LFA-1的机理传感器功能。 DBP＃5专注于 癌细胞的异质性。 DBP＃6和＃7靶向血小板的机械学反应。最后，DBP＃8 ＃9解决了机械在焦点建议中作用的基本问题。我们的原型TDP 技术提供了以与之相同的轻松和简单性测量分子力的方法 免疫染色，流式细胞仪，PCR和ELISA。但是与这些主流技术不同，机械成像， Mechano-PCR，Mechano-Flow和Mechano-Elisa未商业化。因此，试剂和表面 准备协议和数据分析例程必须由最终用户定制。这可以 对非专家的挑战，不是例行公事。因此，CPMM将整合一个强大的社区 参与（CE）组件。 CE活动将集中于动手培训研讨会，方法发布 文章，虚拟节目系列，行业参与，强大的网络影响力和三个关键 机械生物学会议将有助于加速采用张力探针技术。这些CE 活动最终将导致商业化，这将使各个方面的广泛传播传播 细胞生物学群落。