Optical probe and instrumentation development for in vivo near-infrared fluorescence imaging of Alzheimer's disease

用于阿尔茨海默病体内近红外荧光成像的光学探针和仪器开发

• 批准号: 10554311
• 负责人: Steven Shuyu Hou
• 金额: $ 12.73万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-02-01 至 2027-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10554311
• 关键词: APP-PS1; Affinity; Alzheimer' s Disease; Alzheimer' s disease model; Alzheimer' s disease pathology; Alzheimer’s disease biomarker; Amyloid; Amyloid beta-Protein; Animals; Antibodies; Binding; Biodistribution; Blood Vessels; Brain; Chemical Structure; Custom; DAP kinase; Dependence; Deposition; Derivation procedure; Development; Disease Progression; Drug Kinetics; Dyes; Early Diagnosis; Environment; Enzymes; Fluorescence; Fluorescent Dyes; Fluorescent Probes; General Hospitals; Human; Image; Impaired cognition; In Vitro; Individual; International; Label; Leadership; Libraries; Magnetic Resonance Imaging; Massachusetts; Measures; Memory impairment; Mentored Research Scientist Development Award; Mentors; Methods; Microscope; Microscopic; Modeling; Monitor; Multi-modal optical imaging; Mus; Near-infrared optical imaging; Neurodegenerative Disorders; Neurofibrillary Tangles; Nonionizing Radiation; Optics; Pathologic; Pathway interactions; Patients; Penetration; Performance; Photons; Positron-Emission Tomography; Property; Quantitative Structure-Activity Relationship; Research; Research Personnel; Resolution; Resources; Scalp structure; Senile Plaques; Skin; Structure; System; Techniques; Testing; Therapeutic; Therapeutic Effect; Time; Tissues; Training; Transgenic Mice; Validation; Work; abeta accumulation; blood-brain barrier crossing; candidate identification; cost; cranium; efficacy study; extracellular; falls; fluorophore; imaging approach; imaging biomarker; imaging modality; imaging study; imaging system; in vivo; in vivo evaluation; in vivo imaging; inhibitor; instrumentation; medical schools; microscopic imaging; molecular imaging; mouse model; multi-photon; multimodality; multiphoton microscopy; neurofibrillary tangle formation; neuropathology; non-invasive imaging; non-invasive monitor; non-invasive optical imaging; novel; optical imaging; optical spectra; pre-clinical; protein aggregation; simulation; single photon emission computed tomography; skills; small molecule libraries; tau Proteins; tau aggregation; theranostics; tool; treatment response; tumor

PROJECT SUMMARY / ABSTRACT The main neuropathological hallmarks of Alzheimer's disease (AD) are the extracellular accumulation of amyloid plaque deposits and intracellular formation of neurofibrillary tangles (NFTs). To enable the study of disease progression and the effect of therapeutics in preclinical mouse models of AD, there is an urgent need for non- invasive methods of imaging both hallmarks in the living brain. The most established non-invasive imaging approaches at the whole animal level are magnetic resonance imaging (MRI), positron emission tomography (PET), and single photon emission computed tomography (SPECT). However, only optical techniques like near- infrared (NIR) fluorescence imaging have the theoretical spatial resolution to image individual amyloid plaques and NFTs. In contrast to PET and SPECT, NIR fluorescence has the additional advantage of using safe, non- ionizing radiation. Previous NIR fluorescence probes for AD have mostly emitted in the 600-650 nm wavelength range. While these probes can be used to provide low resolution, bulk estimates of AD pathology burden in vivo and to distinguish between AD mice and wildtype controls, they lack the ability to provide absolute quantitation and cannot be used to image non-invasively with microscopic resolution. To enable non-invasive optical imaging through the intact scalp and intact skull of mice, the wavelength of the fluorescent probe needs to be shifted into the NIR-II spectrum (1,000-1,700 nm). At these longer wavelengths, the penetration depth through tissue is increased dramatically while the background caused by intrinsic autofluorescence is minimized. Here we propose to develop and identify candidate probes for NIR-II imaging by employing two complimentary strategies. In the first approach, we will modify the chemical structure of a promising candidate probe, with existing affinity for amyloid plaques and NFTs to enhance its binding affinity and NIR-II fluorescence emission. In the second approach, we will screen an existing chemical library of > 650 NIR fluorophores using chemical structure-activity modeling and in vitro and in vivo characterization. To image our candidate NIR-II fluorophores in vivo, we will build a custom multi-modal microscope combining NIR confocal and multiphoton microscopy and fully characterize its performance with simulations and experimental validation. We will conduct longitudinal, in vivo imaging studies in AD mice to validate our developed probes and instrumentation and demonstrate for the first time non-invasive monitoring of AD pathology with microscopic resolution. The excellent scientific environment and world-class resources provided by the Massachusetts General Hospital and Harvard Medical School, as well as the internationally recognized leadership of the applicant's mentors, along with the expertise of the proposed collaborators, will be key to the successful completion of the proposed research. This K01 award will be instrumental for the applicant to strengthen his skillset with training in optical probe development and in vivo imaging in AD mice and to take the next steps towards becoming an independent researcher.

项目总结/摘要 阿尔茨海默病（Alzheimer's disease，AD）的主要神经病理学特征是细胞外淀粉样蛋白的积聚 斑块沉积和神经纤维缠结（NFT）的细胞内形成。为了研究疾病 研究进展和治疗剂在AD临床前小鼠模型中的作用，迫切需要非- 在活体大脑中对这两个标志进行成像的侵入性方法。最成熟的非侵入性成像 在整个动物水平的方法是磁共振成像（MRI），正电子发射断层扫描， (PET)和单光子发射计算机断层扫描（SPECT）。然而，只有光学技术，如近- 红外（NIR）荧光成像具有对单个淀粉样蛋白斑块成像的理论空间分辨率 和NFT。与PET和SPECT相比，NIR荧光具有使用安全、非放射性的额外优点。 电离辐射以前用于AD的NIR荧光探针大多在600-650 nm波长下发射 范围虽然这些探针可用于提供体内AD病理负荷的低分辨率、批量估计， 为了区分AD小鼠和野生型对照，他们缺乏提供绝对定量的能力， 并且不能用于以显微分辨率进行非侵入性成像。为了实现非侵入式光学成像， 通过小鼠的完整头皮和完整头骨，荧光探针的波长需要转移到 NIR-II光谱（1000 - 1700 nm）。在这些较长的波长下，通过组织的穿透深度为 显著增加，而由固有自发荧光引起的背景最小化。这里我们 建议通过采用两种互补策略开发和鉴定用于NIR-II成像的候选探针。 在第一种方法中，我们将修改有希望的候选探针的化学结构， 用于淀粉样蛋白斑块和NFT，以增强其结合亲和力和NIR-II荧光发射。在第二 方法，我们将筛选现有的化学库> 650近红外荧光团使用化学结构活性 建模以及体外和体内表征。为了在体内成像我们的候选NIR-II荧光团，我们将 建立一个定制的多模态显微镜结合近红外共焦和多光子显微镜， 通过模拟和实验验证来表征其性能。我们将在体内进行纵向研究 在AD小鼠中进行成像研究，以验证我们开发的探针和仪器，并首次证明 以显微分辨率对AD病理进行时间非侵入性监测。良好的科学环境 以及由马萨诸塞州总医院和哈佛医学院提供的世界级资源， 以及申请人导师的国际公认的领导力，沿着的专业知识， 建议的合作者，将是成功完成拟议研究的关键。K 01奖项将 有助于申请人通过光学探针开发和体内 在AD小鼠中进行成像，并采取下一步措施成为一名独立的研究人员。