Integrated Imaging Tools for Intercellular Chemokine Signalling

用于细胞间趋化因子信号转导的集成成像工具

• 批准号: 10706896
• 负责人: Gary D Luker
• 金额: $ 21.88万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-09 至 2026-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10706896
• 关键词: Automobile Driving; Binding; Biochemistry; Bioluminescence; Breast Cancer Cell; CXCL11 gene; CXCR3 gene; CXCR4 Receptors; CXCR4 gene; Cell Proliferation; Cell surface; Cells; Chemotaxis; Complex; Computer Models; Coupling; Data Set; Drug Targeting; Environment; Event; Exhibits; Extracellular Matrix; Fluorescence Microscopy; Future; G-Protein-Coupled Receptors; Heterogeneity; Human; Image; Image Analysis; Imaging Device; Imaging technology; Immune; Invaded; Investigation; Ligands; Malignant Neoplasms; Measures; Mediator; Methods; Molecular; Neoplasm Metastasis; Organism; Output; Pathway interactions; Patients; Pattern; Performance; Phosphotransferases; Positioning Attribute; Process; Proliferating; Protein Isoforms; Receptor Signaling; Reporter; Research; Research Personnel; Resolution; Role; Shapes; Signal Transduction; Signaling Molecule; Site; Spatial Distribution; Specificity; Stromal Cell-Derived Factor 1; Stromal Cells; System; Technology; Testing; Tumor Biology; angiogenesis; bioluminescence imaging; cancer cell; cancer therapy; cell motility; cell type; cellular imaging; chemokine; chemokine receptor; chemokine therapy; clinical translation; complement system; drug development; extracellular; fluorescence imaging; high dimensionality; image processing; imaging study; improved; innovation; intercellular communication; large scale data; live cell imaging; malignant breast neoplasm; molecular imaging; multiplexed imaging; quantitative imaging; receptor; receptor binding; recruit; response; scavenger receptor; targeted treatment; technology development; therapy design; tool; treatment optimization; tumor

Chemokines and their G-protein coupled receptors (GPCRs) are key mediators of intercellular signaling between cancer cells and multiple stromal cell types. Cancer cell proliferation, local invasion, systemic metastasis, recruitment of immune cells and angiogenesis all are regulated by spatial and temporal dynamics of chemokines. The central role of chemokines in tumor biology has made them an attractive target for therapies designed to inhibit or enhance chemokine signaling. However, much of the complex biochemistry of chemokines remains to be discovered, due to the technical challenges of tracing their presence and function in living systems. Chemokines are effective at signaling at very low levels, and their spatial distribution is complex due to their ability to bind to cell surfaces and extracellular matrix. Furthermore, internalization of chemokines by GPCRs and scavenger receptors shape local chemokine gradients by sequestering and degrading chemokines. Experimental evidence and computational modeling by our lab show that short-range, steep local gradients of chemokine CXCL12 over distances of a single cell most effectively drive signaling and chemotaxis through receptor CXCR4. To enable transformative studies of chemokines and ultimately advance applications of drugs targeting chemokines in cancer, we propose to develop a new suite of quantitative molecular imaging tools to measure the spatial and temporal dynamics of chemokine distribution, association, and signaling in hundreds of single cells. These tools will allow us to measure cell signaling patterns during chemotaxis and identify potential mechanisms underlying intercellular heterogeneity in responses to chemokines. We will 1) Generate an integrated, multiplexed bioluminescence and fluorescence microscopy toolbox for single-cell imaging of extracellular and intracellular steps in chemokine signaling; and 2) Test our chemokine imaging technology as a generalizable, quantitative approach applicable to patient-derived cells. We will uniquely combine dynamic, single-cell bioluminescence and fluorescence microscopies technologies to measure numerous molecular events cells use to convert chemokine inputs into signaling outputs and chemotaxis. We will couple our multiplexed imaging readouts with advanced image processing methods to generate high-dimensional quantitative data sets needed for basic studies of chemokines and integration with other large-scale data. Our innovative imaging tools will allow an unprecedented view of chemokine gradients and heterogeneity of cellular responses in complex environments. This technology will transform investigations of chemokines in cancer and advance ongoing efforts to target chemokines for therapy.

趋化因子及其G蛋白偶联受体（GPCR）是细胞间的关键介体 癌细胞与多种基质细胞类型之间的信号传导。癌细胞增殖，局部 入侵，全身转移，免疫细胞的募集和血管生成都受到 趋化因子的空间和时间动力学。趋化因子在肿瘤生物学中的核心作用 使它们成为旨在抑制或增强趋化因子的疗法的有吸引力的靶标 信号。但是，许多复杂的趋化因子生物化学仍然有待发现， 由于追踪其在生活系统中的存在和功能的技术挑战。 趋化因子在非常低的信号传导方面有效，它们的空间分布很复杂 由于它们与细胞表面和细胞外基质结合的能力。此外，内在化 GPCR和清道夫受体的趋化因子塑造局部趋化因子梯度 隔离和降解趋化因子。实验证据和计算建模 我们的实验室表明，趋化因子CXCL12的短距离，陡峭的局部梯度在A 单细胞最有效地通过受体CXCR4驱动信号传导和趋化性。启用 趋化因子的变革性研究，并最终提高靶向药物的应用 我们建议开发一套定量分子成像工具的趋化因子，我们建议开发一套新的套件 测量趋化因子分布，关联和 在数百个单元中发出信号。这些工具将使我们能够测量细胞信号模式 在趋化性期间，并确定细胞间异质性的潜在机制 对趋化因子的反应。我们将1）产生一种集成的，多重的生物发光和 荧光显微镜工具箱，用于细胞外和细胞内步骤的单细胞成像 趋化因子信号传导； 2）测试我们的趋化因子成像技术是一种可推广的， 适用于患者衍生细胞的定量方法。我们将独特地结合动态， 单细胞生物发光和荧光显微镜技术，以测量众多 分子事件细胞用于将趋化因子输入转化为信号输出和趋化性。 我们将将我们的多路复用成像读数与高级图像处理方法相结合到 生成趋化因子基础研究所需的高维定量数据集和 与其他大规模数据集成。我们的创新成像工具将允许空前 复杂环境中细胞反应的趋化因子梯度和异质性的视图。 这项技术将改变对癌症趋化因子的研究，并进步 旨在靶向趋化因子进行治疗的努力。