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.

项目概要 拟议的分子机械技术探针中心（CPMM）将致力于开发和优化 能够研究活细胞中的机械生物学和机械传导途径的技术。这 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 Mattheyses、Khalid Salaita 和 Yonggang Ke 在联合出版和工作方面有着良好的记录 共同开发张力探针技术。在 TDP#1：机械生物学的高分辨率探针中，我们 将创造出“坚不可摧”的探针，可以突破 DNA 张力的空间和方向分辨率的极限 探针技术。 Tension-PAINT 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，以绘制在机械附近组装的蛋白质的图谱 活性受体。将使用交钥匙荧光偏振方法来绘制力方向 商业显微镜。在 TDP#2：机械标记探针中，我们将开发力诱导方法 标记。核心设计特征是 DNA 探针，可在以下位置介导结合事件或解离事件： 力的阈值水平。根据机械事件的幅度和频率对细胞进行标记 由细胞表面受体产生。该 TDP 将导致使用流式细胞术对细胞进行高通量分析 还将允许蛋白质组学分析打开“力学组学”的大门。 TDP#3 下：力量增强 传感器，这里的核心技术是放大机械输入的响应性 DNA 结构。 CPMM 有九个相关的首个驱动生物医学项目 (DBP)，由来自不同地域的团队领导 合作者。 DBP #1-#4 专注于 T 细胞的力学生物学，并使用 CPMM 工具来测试 T 细胞受体 (TCR) 和粘附受体 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 针对血小板的机械传感器反应。最后，DBP#8 和＃9解决了力学在粘着斑中的作用的基本问题。我们的原型 TDP 技术提供了与测量分子力相同的轻松和简单的方法 免疫染色、流式细胞术、PCR 和 ELISA。但与这些主流技术不同的是，机械成像 mechano-PCR、mechano-flow 和 mechano-ELISA 尚未商业化。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 对非专家来说具有挑战性，而且不是常规的。因此，CPMM 将整合一个强大的社区 参与 (CE) 组件。 CE 活动将侧重于实践培训研讨会、方法出版 文章、虚拟研讨会系列、行业参与、强大的网络存在以及三个关键的参与 机械生物学会议将有助于加速张力探针技术的采用。这些CE 活动最终将导致商业化，从而能够在各个领域广泛传播 细胞生物学界。