NEAR INFRARED PH-SENSITIVE MOLECULAR PROBES FOR MICROSCOPY OF CELLS AND TISSUES

用于细胞和组织显微镜检查的近红外 PH 敏感分子探针

• 批准号: 7373077
• 负责人: Samuel Achilefu
• 金额: $ 29.66万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-04-01 至 2012-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7373077
• 关键词: Acidosis; Animals; Antibodies; Biochemical; Biological; Biological Assay; Capillary Electrophoresis; Cell Cycle; Cell Maintenance; Cell Proliferation; Cell membrane; Cell surface; Cells; Cellular biology; Class; Condition; Confocal Microscopy; Depth; Detection; Disease; Dyes; Environment; Enzymes; Event; Fluorescence; Fluorescent Dyes; Generations; Glean; Goals; Growth; Human; Image; Incubated; Integrins; Invasive; Label; Lead; Life; Ligands; Light; Localized; Lysosomes; MCF7 cell; Magnetic Resonance Imaging; Magnetic Resonance Spectroscopy; Maps; Measures; Methodology; Methods; Microelectrodes; Microscopy; Molecular; Molecular Probes; Monitor; Mus; Nature; Organelles; Pathologic; Pathologic Processes; Penetration; Peptides; Phenotype; Phototoxicity; Physiological; Physiological Processes; Positioning Attribute; Process; Property; Protein Conformation; Purpose; Quantitative Microscopy; Radiation; Range; Research; Research Personnel; Resistance; Serous; Signal Transduction; Solid Neoplasm; Spectrum Analysis; Standards of Weights and Measures; Sum; Technology; Tissue Sample; Tissues; Translating; Translations; Tumor Tissue; Visible Radiation; absorption; base; biomaterial compatibility; body system; cancer cell; cellular imaging; cyanine dye; cytotoxic; cytotoxicity; design; diffuse optical tomography; fluorophore; functional status; in vivo; malignant breast neoplasm; method development; molecular imaging; optical imaging; pH gradient; receptor; receptor mediated endocytosis; response; trafficking; transmission process; tumor; tumor progression

DESCRIPTION (provided by applicant): Many physiological processes function efficiently within a well-controlled pH range and significant deviations from this range typically are indicative of pathologic conditions. Within cells, maintenance of a pH gradient in different organelles allows for differential activation of certain enzymes or generation of active conformations of proteins for transient or prolonged activity. Globally, the sum of cellular events can reflect on the pH of tissues and organ systems. A common strategy to monitor the pH of biochemical events relies on the use of pH-sensitive fluorescent molecular probes to monitor diverse physiological and pathological processes, including studies related to cancer, cell proliferation, endocytotic and other physiological processes. Currently, there is a major disconnection between the use of pH-sensitive fluorescent dyes in molecular cell biology and for imaging diseases in vivo. This is because most of these dyes fluoresce in the visible wavelengths where absorption of light by native fluorophores and autofluorescence limits their use in both tissue and cell imaging. To overcome these problems, we propose to develop new classes of pH-sensitive near infrared fluorescent molecular probes that are useful for imaging molecular processes by microscopy and whole-body animal imaging by diffuse optical tomography. Advantages of this approach include high detection sensitivity because of low autofluorescence, low phototoxicity because of the use of low energy radiation, and the extension of the spectral window to the 900 nm which allows the use of multiple fluorophores between 400 and 900 nm wavelengths. More importantly, information gleaned from the molecular cell biology studies can be translated to small animals, and eventually to humans with the same pHsensitive NIR molecular probe because NIR light can penetrate into deep tissues. We will accomplish these goals by (1) designing and developing two new classes of pHsensitive NIR dyes; (2) demonstrating their utility to image pH-dependent molecular processes in cells; and (3) imaging the onset of acidosis in solid tumors in vivo. The products generated from this project will be distributed to other investigators for research purposes. Project Narrative The goals of this project are to (1) design and develop two new classes of pH-sensitive NIR dyes; (2) demonstrate their utility to image pH-dependent molecular processes in cells by microscopy; and (3) image the onset of acidosis in solid tumors in vivo. The technology platform developed will be useful for diverse pH-based biological applications.

描述（由申请人提供）：许多生理过程在良好控制的 pH 范围内有效地发挥作用，并且与该范围的显着偏差通常表明病理状况。在细胞内，不同细胞器中 pH 梯度的维持可实现差异化 某些酶的激活或蛋白质活性构象的产生，以实现短暂或长期的活性。在全球范围内，细胞事件的总和可以反映组织和器官系统的 pH 值。监测生化事件 pH 值的常用策略依赖于使用 pH 敏感荧光分子探针来监测各种生理和病理过程，包括与癌症、细胞增殖、内吞和其他生理过程相关的研究。目前，pH 敏感荧光染料在分子细胞生物学中的使用与体内疾病成像之间存在重大脱节。这是因为大多数这些染料在可见波长范围内发出荧光，其中天然荧光团对光的吸收和自发荧光限制了它们在组织和细胞成像中的使用。为了克服这些问题，我们建议开发新型 pH 敏感近红外荧光分子探针，这些探针可用于通过显微镜对分子过程进行成像以及通过扩散光学断层扫描对动物全身进行成像。这种方法的优点包括由于低自发荧光而具有高检测灵敏度、低光毒性 由于使用低能辐射，并且光谱窗口延伸至 900 nm，从而允许使用 400 至 900 nm 波长之间的多种荧光团。更重要的是，从分子细胞生物学研究中收集的信息可以转化为小动物，并最终利用相同的 pH 敏感近红外分子探针转化为人类，因为近红外光可以穿透深层组织。我们将通过以下方式实现这些目标：(1) 设计和开发两类新型 pH 敏感型近红外染料； (2) 展示它们在细胞中对 pH 依赖性分子过程进行成像的实用性； (3)体内实体瘤酸中毒发作的成像。该项目产生的产品将分发给其他研究人员用于研究目的。项目叙述 该项目的目标是 (1) 设计和开发两类新的 pH 敏感型近红外染料； (2) 展示它们通过显微镜对细胞中 pH 依赖性分子过程进行成像的实用性； (3)对体内实体瘤酸中毒的发生进行成像。开发的技术平台将可用于各种基于 pH 的生物应用。