Photocleavable Mass-Tags for Spatial Multiomics of Alzheimer’s Brain Tissue

用于阿尔茨海默病脑组织空间多组学的光裂解质量标签

• 批准号: 10483988
• 负责人: Mark Lim
• 金额: $ 135.5万
• 依托单位: AMBERGEN, INC
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10483988
• 关键词: AD transgenic mice; Address; Aducanumab; Affect; Agar; Alzheimer' s Disease; Alzheimer' s disease brain; Alzheimer' s disease pathology; Alzheimer' s disease therapy; Amyloid beta-Protein; Antibodies; Autopsy; Big Data; Biological Markers; Biotechnology; Boston; Brain Diseases; Brain imaging; Cholesterol; Chronic; Collaborations; Data Analyses; Dementia; Diagnostic; Drug Targeting; FDA approved; Fluorescence; Functional disorder; Goals; Health Care Costs; Hospitals; Human; Image; Image Analysis; Imaging Techniques; Imaging technology; Immunohistochemistry; Institutes; Label; Lectin; Letters; Link; Lipids; Machine Learning; Malignant neoplasm of prostate; Maps; Methods; Modeling; Molecular; Molecular Disease; Molecular Target; Monoclonal Antibodies; Multiomic Data; Mus; Mutation; Neurodegenerative Disorders; Neurons; Peptides; Persons; Pharmaceutical Preparations; Pharmacologic Substance; Phase; Phospholipids; Physics; Play; Polysaccharides; Procedures; Process; Production; Proteins; Public Health; Research; Research Personnel; Resolution; Role; Sampling; Senile Plaques; Source; Spatial Distribution; Specimen; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Tissue imaging; Tissues; Transgenic Mice; Uncertainty; Universities; Validation; Woman; base; brain tissue; cohort; commercialization; design; drug candidate; human disease; human old age (65+); innovation; instrument; malignant breast neoplasm; mass spectrometric imaging; molecular imaging; molecular pathology; mouse model; multiple omics; novel; operation; peptide drug; polypeptide; presenilin-1; receptor; small molecule; tau Proteins

Summary/Abstract Alzheimer's Disease (AD) is a chronic neurodegenerative disorder characterized by progressively worsening dementia. AD currently affects over 6.2 million persons in the U.S. and approximately 30 million world-wide with 70% over the age of 65. The total public health cost of AD is expected to reach over $20 trillion by 2050. Despite extensive efforts to develop AD therapies including small molecules, monoclonal antibodies and peptide-based drugs, only aducanumab, whose efficacy is in doubt, has been approved by the FDA since 2004. A major challenge is elucidating the molecular pathology involved in AD in order to develop effective early diagnostics and drugs. While amyloid plaque formation due to aggregation of different Aβ-peptides has been an important focus, a myriad of other molecules including tau, neuronal and glial receptors, endosomal-lysosomal related proteins, glycans, phospholipids, cholesterol and metabolites have also been implicated in AD pathology. In order to obtain a detailed understanding of the possible role of these diverse molecular species as well as the molecular targeting of candidate drugs, there is an urgent need to develop sufficiently powerful, highly multiplexed and multiomic tissue imaging techniques that can map at cellular resolution the 2D-spatial distribution and association of these diverse, AD molecular players. The proposed Phase II project seeks to address this challenge by applying a new highly- multiplexed, targeted method termed mass spectrometric imaging immunohistochemistry (MSI-IHC™). MSI- IHC™ is based on the use of novel photocleavable mass-tags (PC-MTs) developed by AmberGen which when linked to antibody or lectin probes enable targeted biomolecules to be identified in the mass spectrometric image. This approach significantly exceeds the multiplex capability of fluorescence immunohistochemistry (IHC) and previous cleavable mass-tag based methods which are generally limited to 5 biomarkers or require extensive cycling procedures in the case of fluorescence. In addition, the ability to combine MSI-IHC™ with label-free, untargeted small molecule mass spectrometric imaging (MSI) as well as fluorescence IHC imaging, on the same sample, greatly extends its power. This is possible using unique double-labeled fluorescent-PC-MT probes and performing 2 rounds of MSI. Together, these innovations can provide a much more comprehensive multiomic picture of the role of various molecules in AD pathology. In Phase I, we have demonstrated the feasibility of this approach on mouse and human brain tissue specimens including the ability to image simultaneously a variety of AD related molecules. In Phase II we will build on this progress by applying MSI-IHC™ to human and transgenic AD mouse brain tissue obtained from collaborators and commercial sources. One goal, in collaboration with Prof. R.A. Nixon at NYU, a leading AD researcher, will be to investigate the role of neuronal endosomal dysfunction, the earliest known pathobiology specific to AD. Image analysis with be performed using novel statistical physics and AI methods previously developed for AD tissue and brain imaging.

摘要/摘要 阿尔茨海默氏病（AD）是一种慢性神经退行性疾病，其特征是逐渐担心 失智。广告目前影响美国超过620万人，全球约3000万人 65岁以上的70％。到2050年，AD的总公共卫生成本预计将超过20万亿美元。 广泛的努力开发广告疗法，包括小分子，单克隆抗体和基于胡椒的疗法 自2004年以来，只有毒品，只有阿德卡司纳单抗的有效性就得到了质疑。 为了开发有效的早期诊断和药物，阐明了与AD有关的分子病理。 虽然由于不同Aβ肽的聚集而导致的淀粉样菌斑是一个重要的重点，但无数 其他分子，包括tau，神经元和神经胶质受体，内体 - 溶酶体相关蛋白，聚糖， AD病理学也暗示了磷脂，胆固醇和代谢产物。为了获得详细的 了解这些不同分子物种的可能作用以及候选者的分子靶向 药物，迫切需要开发足够强大的，高度多重的和多构的组织成像 可以在细胞分辨率绘制的技术2D空间分布和这些潜水员的关联，AD 分子玩家。拟议的第二阶段项目旨在通过应用新的高度 - 多路复用的靶向方法称为质谱成像免疫组织化学（MSI-IHC™）。 ms IHC™基于使用Ambergen开发的新型光电标记（PC-MT）的使用 对于抗体或讲座问题，可以在质谱图像中识别靶向生物分子。这 方法大大超过了荧光免疫组织化学（IHC）和以前的多重能力 基于可切合的质量标签方法通常限于5个生物标志物或需要大量循环 在荧光的情况下。另外，将MSI-IHC™与无标签，不靶向的能力相结合 小分子质谱成像（MSI）以及荧光IHC成像，在同一样品上 扩展其力量。使用独特的双标签荧光-PC-MT问题并执行2发子弹，这是可能的 MSI。这些创新在一起可以提供更全面的多构图。 AD病理学中的各种分子。在第一阶段，我们已经证明了这种方法在鼠标上的可行性， 人脑组织标本，包括同时对各种AD相关的分子进行图像的能力。在 第二阶段，我们将通过将MSI-IHC™应用于人和转基因AD小鼠脑组织来建立这一进度 从合作者和商业来源获得。一个目标，与R.A.教授合作尼克松在纽约大学 领先的广告研究人员将研究神经元内体功能障碍的作用，最早已知 特定于AD的病理生物学。使用新型统计物理和AI方法进行图像分析 先前用于AD组织和脑成像。