Dinuclear ruthenium light-switches as multi-output sub-cellular imaging probes within live cells and tissues

双核钌光开关作为活细胞和组织内的多输出亚细胞成像探针

• 批准号: EP/M015572/1
• 负责人: James Thomas
• 金额: $ 90.72万
• 依托单位: University of Sheffield
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2015
• 资助国家: 英国
• 起止时间: 2015 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=EP%2FM015572%2F1
• 关键词: Dinuclear; ruthenium; light; switches; multi

Just as atoms are the basic unit of matter, cells are the basic unit of life. All the functions required to maintain a healthy organism can ultimately traced back to molecular processes occurring within cells. When there is a malfunction of these processes or they are disrupted, disease states, including cancer, can arise. To understand the complex structures and functions of cellular components in more depth, cell biologists wish to probe cells at a molecular level. However, since cell are mostly transparent and colourless, coloured or luminescent stains must be used to mark and visulise specific cellular components. In previous work the Thomas group has identified a compound that is a luminescent probe for DNA (the genetic "blueprint" molecule) within the nucleus of cells. This probe is particularly exciting as, unlike commercial cells, its emission is induced by illumination with low energy light, which can penetrate into tissue through layers of cells and is not deleterious to live samples. Furthermore once "lit up" the bound probe emission lifetime is also a distinctive marker for DNA. This is significant as this lifetime marker can be used as a "fingerprint" , even if emission from other naturally occurring molecules within the cell is occurring,nIn this project these exciting results will be further developed.The probe we used in our original studies contains two chiral metal centres (non-superimposible "mirror images") that were not resolved, consequently it is a mixture of products. Since many biomolecules are also chiral, and binding between molecules can be highly dependent on the chirality of each component, in this project we will carry out studies on chirally pure examples of the original probes to investigate whether the individual stereoisomers are taken up and bind/image different cellular targets. We will also make a series of related probes designed to bind to different in cellulo targets. While optical microscopy is an attractive tecnique for dynamic imaging, it relies on a probe emitting light when bound to a target, which is not always the case. Furthermore other techniques - such as electron microscopy - can potentially provide (static) imaging at a higher resolution. Consequently, the use of the new compounds as multifunctional probes will also be investigated. In particular their use as probes for Transmission Electron Microscopy and Raman Microscopy techniques will also be pursued. Such probes will be useful as imaging at a range of scales using the same probe will be possible and, potentially, systems that can image separate structures through different modalities will be produced.The compounds we have identified do not passively diffuse into cells, but are actively taken up. Very interestingly, we have found that although they are taken up by most of the commonly used cell lines used in biological and medical research, not all lines take up the probes. To exploit this striking result, in a proof-of-concept study we will investigate "tumour genesis" in a 3-D skin tumour model. A successful outcome in this study will help understand the process of tumour development and may lead to new diagnostic technologies.

正如原子是物质的基本单位一样，细胞是生命的基本单位。维持健康有机体所需的所有功能最终都可以追溯到细胞内发生的分子过程。当这些过程出现故障或被破坏时，可能会出现包括癌症在内的疾病状态。为了更深入地了解细胞成分的复杂结构和功能，细胞生物学家希望在分子水平上探测细胞。然而，由于细胞大多是透明且无色的，因此必须使用有色或发光染色剂来标记和可视化特定的细胞成分。在之前的工作中，托马斯小组已经鉴定出一种化合物，它是细胞核内 DNA（遗传“蓝图”分子）的发光探针。该探针特别令人兴奋，因为与商业细胞不同，它的发射是通过低能光照射诱导的，可以穿过细胞层渗透到组织中，并且对活样品无害。此外，一旦“点亮”，结合探针的发射寿命也是 DNA 的独特标记。这是很重要的，因为这个寿命标记可以用作“指纹”，即使细胞内其他天然存在的分子正在发生发射，在这个项目中，这些令人兴奋的结果将得到进一步发展。我们在最初的研究中使用的探针包含两个手性金属中心（非重叠的“镜像”）未解析，因此它是产物的混合物。由于许多生物分子也是手性的，分子之间的结合可能高度依赖于每个组分的手性，在这个项目中，我们将对原始探针的手性纯实例进行研究，以研究单个立体异构体是否被吸收并结合/对不同的细胞目标进行成像。我们还将制作一系列相关探针，设计用于结合不同的细胞靶标。虽然光学显微镜对于动态成像来说是一种有吸引力的技术，但它依赖于与目标结合时发射光的探针，但情况并非总是如此。此外，其他技术 - 例如电子显微镜 - 可以提供更高分辨率的（静态）成像。因此，还将研究新化合物作为多功能探针的用途。特别是，还将继续将它们用作透射电子显微镜和拉曼显微镜技术的探针。此类探针将非常有用，因为使用同一探针在一系列尺度上进行成像将是可能的，并且可能会产生能够通过不同方式对单独结构进行成像的系统。我们已经识别出的化合物不会被动扩散到细胞中，而是会通过积极采取。非常有趣的是，我们发现虽然它们被生物和医学研究中使用的大多数常用细胞系所吸收，但并非所有细胞系都吸收探针。为了利用这一惊人的结果，在概念验证研究中，我们将研究 3D 皮肤肿瘤模型中的“肿瘤发生”。这项研究的成功结果将有助于了解肿瘤发展的过程，并可能带来新的诊断技术。

项目成果

A three-in-one-bullet for oesophageal cancer: replication fork collapse, spindle attachment failure and enhanced radiosensitivity generated by a ruthenium(ii) metallo-intercalator.

• DOI: 10.1039/c7sc03712k
• 发表时间: 2018-01-28
• 期刊: Chemical science
• 影响因子: 8.4
• 作者: Gill MR;Jarman PJ;Halder S;Walker MG;Saeed HK;Thomas JA;Smythe C;Ramadan K;Vallis KA
• 通讯作者: Vallis KA

A Dinuclear Osmium(II) Complex Near-Infrared Nanoscopy Probe for Nuclear DNA

• DOI: 10.1021/jacs.1c10325
• 发表时间: 2021-12-08
• 期刊: JOURNAL OF THE AMERICAN CHEMICAL SOCIETY
• 影响因子: 15
• 作者: Droge, Fabian;Noakes, Felicity F.;Thomas, Jim A.
• 通讯作者: Thomas, Jim A.

A Dinuclear Ruthenium(II) Complex Excited by Near-Infrared Light through Two-Photon Absorption Induces Phototoxicity Deep within Hypoxic Regions of Melanoma Cancer Spheroids

• DOI: 10.1021/jacs.9b11313
• 发表时间: 2020-03-11
• 期刊: JOURNAL OF THE AMERICAN CHEMICAL SOCIETY
• 影响因子: 15
• 作者: Raza, Ahtasham;Archer, Stuart A.;Haycock, John W.
• 通讯作者: Haycock, John W.