Molecular Probes for Biomembrane Recognition

用于生物膜识别的分子探针

• 批准号: 10116431
• 负责人: BRADLEY D. SMITH
• 金额: $ 39.29万
• 依托单位: UNIVERSITY OF NOTRE DAME
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-04-01 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10116431
• 关键词: Animal Cancer Model; Animal Disease Models; Cells; Clinical; Collaborations; Diagnostic; Drug Kinetics; Dyes; Endosomes; Ensure; Fluorescence; Fluorescent Dyes; Fluorescent Probes; Glaucoma; Health Status; Image; Image Enhancement; Imaging Techniques; Label; Light; Lipids; Medical; Methods; Microscopy; Molecular; Molecular Probes; Oligonucleotide Probes; Oligonucleotides; Operative Surgical Procedures; Pathology; Patient imaging; Peptides; Performance; Procedures; Process; Property; Proteins; Protocols documentation; Reporting; Research; Research Personnel; Resolution; Science; Structure; Techniques; Time; Translation Process; Vendor; Virus Diseases; Work; base; bioimaging; clinical imaging; clinical translation; contrast imaging; cyanine; design; exosome; fluorescence imaging; fluorescence-guided surgery; implantable device; improved; in vivo; in vivo imaging; innovation; interest; nanoscale; near infrared dye; polyhistidine; pre-clinical; preclinical imaging; single molecule; trafficking

Project Summary/Abstract Fluorescence imaging is having a transformative impact on general medical science with the advent of powerful preclinical techniques such as super-resolution microscopy and single-molecule-tracking, along with emerging clinical procedures such as fluorescence guided surgery and rapid pathology. Each method requires a specific type of fluorescent molecular probe, and in many cases non-optimal probe performance is a major factor that is limiting long term progress and broad impact. Thus, the central objective of this research is to develop greatly improved fluorescent dyes and convert them into molecular probes for important applications in preclinical and clinical imaging. The largest set of projects are based on new fluorescent cyanine heptamethine (Cy7) dyes that emit near-infrared (NIR) light with wavelengths >700 nm and enable imaging of living subjects in the wavelength windows known as NIR I and NIR II (the latter window is also called Short Wavelength Infrared). The new Cy7 dyes have innovative molecular design features that greatly enhance image contrast and permit rapid fine-tuning of probe pharmacokinetic profiles. The near-infrared dyes will be converted into various targeted and bioresponsive molecular probes for: (a) preclinical microscopy, including single-molecule localization microscopy, (b) in vivo fluorescence imaging of animal models of disease, and clinical imaging of patients undergoing surgery, (c) long-term implantable devices that use NIR fluorescence to report health status, and (d) biomedical diagnostics and sequencing methods. Some of these Cy7 dyes will be so stable that they can be incorporated, for the first time, into automated peptide and oligonucleotide synthesis protocols. This will have a major impact on clinical translation since it will allow researchers to reliably make their own Cy7 labeled peptide or oligonucleotide probes on a large scale using a standard synthesizer machine. A second set of projects will develop a new class of fluorescent molecules whose unusual self-threaded structures are reminiscent of entangled peptides. The probes have rigid peptide loops that promote outstanding stability and targeting properties, along with bright deep-red fluorescence that is perfectly suited for advanced intracellular microscopy. An early focus is on molecular probes that spontaneously permeate into living cells and permit selective fluorescence labeling of any intracellular protein containing a polyhistidine sequence (His-tagged protein). The work will create the first deep-red fluorescent molecular probes for rigorous nanoscale imaging of subcellular polar lipids during biomedically important dynamic processes such as viral infection and endosome/exosome trafficking. The utility and versatility of these new fluorescent probes will be demonstrated by collaborations with experts in advanced microscopy. Close coordination with a commercial vendor will ensure that the important fluorescent probes developed by this research quickly become available to all interested investigators.

项目总结/摘要 随着荧光成像技术的出现， 强大的临床前技术，如超分辨率显微镜和单分子跟踪，沿着 新兴的临床程序，如荧光引导手术和快速病理学。每种方法都需要 一种特定类型的荧光分子探针，在许多情况下，非最佳的探针性能是一个主要的 限制长期进展和广泛影响的因素。因此，本研究的中心目标是 开发出大大改进的荧光染料，并将其转化为分子探针， 临床前和临床成像。最大的一套项目是基于新的荧光菁七甲川 (Cy7)发射波长>700 nm的近红外（NIR）光并能够对活体成像的染料 在被称为NIR I和NIR II的波长窗口中（后一窗口也被称为短波长 红外线）。新型Cy 7染料具有创新的分子设计特征，可大大增强图像对比度， 允许探针药代动力学曲线的快速微调。近红外染料将被转化为各种 靶向和生物响应分子探针，用于：（a）临床前显微镜检查，包括单分子 定位显微术，（B）疾病动物模型的体内荧光成像，和 接受手术的患者，（c）使用NIR荧光报告健康状况的长期植入式装置， 和（d）生物医学诊断和测序方法。其中一些Cy 7染料非常稳定，以至于 可以第一次整合到自动化肽和寡核苷酸合成方案中。这将 对临床翻译产生重大影响，因为它将使研究人员能够可靠地使他们自己的Cy 7标记 使用标准合成仪大规模制备肽或寡核苷酸探针。第二组 项目将开发一类新的荧光分子，其不寻常的自螺纹结构是 让人联想到纠缠的肽。探针具有刚性肽环，可促进出色的稳定性， 靶向特性，沿着明亮的深红色荧光，非常适合高级细胞内 显微镜早期的重点是分子探针，自发渗透到活细胞中， 选择性荧光标记含有多组氨酸序列（His标记的）的任何细胞内蛋白质 蛋白质）。这项工作将创建第一个深红色荧光分子探针，用于严格的纳米级成像， 亚细胞极性脂质在生物医学上重要的动态过程，如病毒感染， 内体/外体运输。这些新的荧光探针的实用性和多功能性将被证明 通过与先进显微镜专家的合作。与商业供应商密切协调， 这项研究开发的重要荧光探针很快就可以被所有感兴趣的人使用。 investigators.