Developing fluorescent probes for the endogenous gaseous transmitters NO and H2S

开发内源性气体递质 NO 和 H2S 荧光探针

• 批准号: 7872197
• 负责人: Michael Pluth
• 金额: $ 8.92万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-04-01 至 2012-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7872197
• 关键词: Address; Adsorption; Alzheimer' s Disease; Binding; Binding Sites; Biological; Biological Process; Biology; Carbon Nanotubes; Cardiovascular system; Catalysis; Cells; Chemistry; Complex; Cytosol; Detection; Development; Dextrans; Doctor of Philosophy; Down Syndrome; Electromagnetics; Electron Transport Complex III; Electronics; Electrons; Endocytosis; Excision; Family; Fluorescence; Fluorescent Probes; Goals; Human body; Huntington Disease; Hydrogen Sulfide; Hydrophobicity; Hypertension; Image; Immune; In Vitro; Individual; Inflammation; Iron; Joints; Laboratories; Lead; Life; Ligands; Measurement; Mentors; Metals; Methodology; Methods; Microscopic; Monitor; Multiple Sclerosis; Natural regeneration; Nerve Degeneration; Nervous system structure; Nitric Oxide; Nitrogen; Optics; Oxidation-Reduction; Parkinson Disease; Pathway interactions; Penetration; Phase; Photobleaching; Play; Polymers; Positron-Emission Tomography; Postdoctoral Fellow; Process; Production; Property; Reaction; Research; Research Personnel; Resistance; Resolution; Role; Ruthenium; Signal Transduction; Stimulus; Time; Tissues; Transition Elements; United States National Institutes of Health; Vasodilation; Water; Work; base; blood pressure regulation; carcinogenesis; design; dextran; extracellular; fluorophore; functional group; in vivo; interest; metal complex; molecular recognition; nervous system disorder; numb protein; physical property; prevent; public health relevance; scaffold; sensor; single walled carbon nanotube; small molecule; tool; tumor growth

DESCRIPTION (provided by applicant): The candidate received his Ph.D. from the UC Berkeley, under the joint direction of Kenneth Raymond and Robert Bergman where he studied host-guest chemistry, molecular recognition, and catalysis in water-soluble supramolecular complexes. He is currently an NIH postdoctoral fellow in Stephen Lippard's laboratory at MIT working on developing fluorescent probes for nitric oxide. The candidate's research interests span the field of molecular recognition with a specific focus on how microscopic processes lead to the recognition of individual atoms, functional groups and molecules. The candidate will use his background in mechanistic studies and molecular recognition to pursue his research interests as a principle investigator. His independent research will focus on the development of new tools for the detection and imaging of small molecules in biology. Nitric oxide (NO) and hydrogen sulfide (H2S) are now accepted as biologically important gaseous transmitters. Both NO and H2S are produced endogenously and are finely regulated by the body. Nitric oxide is beneficial for vasodilation and immune activity at low cellular concentrations but overproduction can lead to the proliferation of reactive NO species that have been implicated in carcinogenesis and several degenerative neurological disorders, including Alzheimer's (AD), Parkinson's, and Huntington's disease, as well as multiple sclerosis. Similarly, H2S has been implicated in AD, Downs syndrome and other forms of metal deficiency. H2S also plays an active role in inflammation and in blood pressure regulation. Despite the recognized importance of both of these gaseous transmitters, the current methods for detection in live cells are limited. The postdoctoral phase of the proposed research will focus on the development of new NO-selective fluorescent probes that address current limitations of NO detection. Transition metal based NO binding sites will be used to develop probes that can reversibly bind NO and probes that emit in the NIR. The proposed family of fluorescent probes will use paramagnetic (S=1/2) metals serving the dual role as both fluorescence quencher and NO binding site. Coordination of NO will form a diamagnetic (S=0) complex and restore the fluorescence of the pendant fluorophore. Adsorption or covalent attachment of such complexes to solubilized single-walled carbon nanotubes (SWNTs) will be used to develop NO-selective probes that emit in the NIR. The independent research phase of the proposed research will investigate the design of H2S-selective fluorescent probes for the imaging of endogenously produced H2S. Currently, such H2S detection methods are lacking and most measurements rely on bulk tissue measurements. The new H2S-selective fluorescent probes for use in live cells will provide much needed tools for the study of the biological functions of H2S. The unique physical properties of H2S will all be exploited in the design of H2S-selective fluorescent probes. Fluorophores will be derivatized with specially designed protecting groups that can only be removed by H2S. Removal of the fluorophore protecting group will restore the fluorescence, thus forming a turn-on probe for H2S. PUBLIC HEALTH RELEVANCE: Both nitric oxide (NO) and hydrogen sulfide (H2S) have been identified as important endogenous gaseous transmitters in the human body and have been implicated in carcinogenesis, hypertension, and several neurological disorders including Alzheimer's disease, Parkinson's disease, Downs syndrome, and multiple sclerosis. Despite this interest, there are currently few methods to detect or image intracellular levels of these small molecule transmitters. This proposal presents the design of fluorescent probes for NO and H2S, which would allow for the selective detection and imaging of these endogenous gasses in live cells.

描述（由申请人提供）：候选人获得了他的博士学位。在肯尼斯·雷蒙德（Kenneth Raymond）和罗伯特·伯格曼（Robert Bergman）的联合指导下，他从加州大学伯克利分校（UC Berkeley）研究了宿主 - 环化学，分子识别和水溶性超分子复合物的催化。他目前是MIT的Stephen Lippard的实验室的NIH博士后研究员，致力于开发一氧化氮的荧光探针。候选人的研究兴趣跨越了分子识别领域，其特定重点是微观过程如何导致单个原子，官能团和分子的识别。候选人将利用他在机械研究和分子认可方面的背景来追求他作为主要研究者的研究兴趣。他的独立研究将着重于开发生物学中小分子的新工具。 一氧化氮（NO）和硫化氢（H2S）现在被接受为具有生物学上重要的气态发射器。 NO和H2均可内源产生，并由人体细节调节。一氧化氮在低细胞浓度下有益于血管舒张和免疫活性，但生产过多会导致反应性无物种的增殖，这些物种与癌变和几种退化性神经系统疾病有关，包括阿尔茨海默氏症（AD），帕金森氏症和亨廷顿疾病，以及亨廷顿氏病，以及多重棘皮动物。同样，H2S与AD，Downs综合征和其他形式的金属缺乏有关。 H2S在炎症和血压调节中也起着积极作用。尽管这两种气态发射器都具有公认的重要性，但当前在活细胞中检测的方法受到限制。 拟议的研究的博士后阶段将重点介绍新的无选择荧光探针的发展，该探针解决了无检测的当前局限性。基于过渡金属的NO结合位点将用于开发可以可逆地结合NO的探针和在NIR中发出的探针。拟议的荧光探针家族将使用顺磁性（S = 1/2）金属，既可以用作荧光淬灭剂又无结合位点。 NO的配位将形成Diamagnetic（S = 0）复合物，并恢复吊坠荧光团的荧光。这种复合物在溶解的单壁碳纳米管（SWNT）上的吸附或共价附着将用于开发NIR中发出的无选择探针。 拟议的研究的独立研究阶段将研究H2S选择性荧光探针的设计，以成像内源产生的H2s。当前，缺乏此类H2S检测方法，大多数测量依赖于散装组织测量。用于活细胞中的新型H2S选择性荧光探针将为研究H2S的生物学功能提供急需的工具。 H2S的独特物理特性都将在H2S选择性荧光探针的设计中被利用。荧光团将通过专门设计的保护组进行衍生化，这些组只能由H2S去除。去除荧光团保护组将恢复荧光，从而形成对H2S的转交探针。 公共卫生相关性：一氧化氮（NO）和硫化氢（H2S）都被确定为人体中重要的内源气态发射器，并且与癌变，高血压和几种神经系统疾病有关，包括阿尔茨海默氏病，包括阿尔茨海默氏病，帕金森氏病，帕克森氏病，下降综合症和多重层库。尽管有这种兴趣，但目前几乎没有检测或图像这些小分子发射器的细胞内水平的方法。该建议介绍了NO和H2S的荧光探针的设计，这将允许对活细胞中这些内源性气体进行选择性检测和成像。