Clusterin glycosylation as diagnostic and prognostic biomarker for Alzheimer's disease

簇蛋白糖基化作为阿尔茨海默病的诊断和预后生物标志物

基本信息

批准号：
10699168
负责人：
DOBRIN NEDELKOV
金额：
$ 38.48万
依托单位：
ISOFORMIX, INC.
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-06-15 至 2025-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10699168
关键词：
Abeta clearance Abeta synthesis Alzheimer&apos s Disease Alzheimer&apos s disease brain Alzheimer&apos s disease pathology Alzheimer&apos s disease risk Alzheimer&apos s disease therapeutic Alzheimer’s disease biomarker Amyloid Fibrils Amyloid beta-42 Amyloid beta-Protein Amyloid beta-Protein Precursor Antibodies Apolipoprotein E Area Attention Binding Biological Assay Biological Markers Blinded Blood - brain barrier anatomy Brain Clinical Data Deposition Detection Development Diagnosis Disaccharides Enzymes Excision Future Genotype Hippocampus Immunoassay Impairment Individual Late Onset Alzheimer Disease Link MALDI-TOF Mass Spectrometry Mass Spectrum Analysis Measures Molecular Chaperones Neuraminidase Pathogenesis Pathway interactions Pattern Performance Phase Phenotype Plasma Play Polysaccharides Prognostic Marker Protein Isoforms Reproducibility Research Risk Role Sampling Senile Plaques Severities Sialic Acids Specificity Structure Technology Testing Therapeutic Up-Regulation Validation Variant Western Blotting apolipoprotein E-3 apolipoprotein E-4 cohort commercialization cost effective cost efficient detection assay diagnostic biomarker diagnostic value extracellular genetic risk factor genome wide association study glycosylation neuroprotection novel prevent prognostic value sugar sulfated glycoprotein 2 tau Proteins therapeutic development trend β-amyloid burden

项目摘要

PROJECT SUMMARY Clusterin is the third most predominant genetic risk factor for late onset Alzheimer’s disease (AD). Clusterin facilitates the transport of soluble amyloid-b (Ab) across the blood-brain barrier. Clusterin binds soluble forms of Ab, preventing their aggregation into amyloid fibrils. However, there is no clear correlation between plasma or CSF clusterin concentration levels and AD, even though there is substantial evidence for upregulation of clusterin in the AD brain and colocalization with Ab plaques. Clusterin is heavily glycosylated, with ~25% of its mass comprised of N-linked glycans. Glycosylation of clusterin is spatially and temporally regulated, and it is important for its function as an extracellular chaperone. Glycosylation of clusterin may play an important role in the interaction with Ab and its clearance. In this Phase I project we propose to develop, optimize and validate a fast and cost-efficient mass spectrometry (MS)-immunoassay for detection of clusterin glycoforms that reveals both intact glycosylated clusterin and its component glycan structures. We will first optimize assay parameters such as antibody amount, sample volumes and dilutions, post-capture rinses, enzymatic release of sialic acids and the full glycans with sialidase and PNGaseF enzymes, and MS acquisition parameters. The assay performance will be validated by testing its robustness, accuracy, specificity, and reproducibility. We will use the assay on a cohort of matched CSF and plasma samples (n=106) from mildly impaired and healthy individuals at risk of AD. Data in the form of intact clusterin mass peaks shifts between plasma and CSF, mass shifts within samples following sialidase and PNGaseF treatment, the relative ratios of clusterin glycoforms, and IDs and ratios of the released glycans, will be compared across the cohort, and correlations with apoE genotype, apoE isoform-specific glycosylation, Aβ42, Tau and pTau will be examined. The findings from the first cohort will be validated with a second cohort (n=100). Our value proposition is a fast and cost-effective assay for analyzing clusterin glycoforms and glycans that could be utilized for evaluating the role of clusterin glycosylation in AD pathogenesis. Clusterin glycosylation could serve as a biomarker for assessing AD risk, or as a target for the development of therapeutics that modify its glycosylation.

项目概要 Clusterin 是晚发性阿尔茨海默病 (AD) 的第三大遗传风险因素。促进可溶性淀粉样蛋白-b (Ab) 穿过血脑屏障的运输，并与可溶性形式的凝聚素结合。 Ab，防止它们聚集成淀粉样原纤维然而，血浆或淀粉样蛋白之间没有明确的相关性。 CSF 聚集蛋白浓度水平和 AD，尽管有大量证据表明聚集蛋白上调在 AD 大脑中，与 Ab 斑块共定位的蛋白被高度糖基化，约占其质量的 25%。簇蛋白的糖基化在空间和时间上受到调节，并且很重要。由于其作为细胞外伴侣的功能，簇蛋白的糖基化可能在该过程中发挥重要作用。与 Ab 的相互作用及其清除。在这个第一阶段项目中，我们建议开发、优化和验证快速且经济高效的质谱法 (MS)-免疫测定法检测凝聚素糖型，揭示完整的糖基化凝聚素及其我们将首先优化抗体量、样品体积等测定参数。和稀释、捕获后漂洗、唾液酸和全聚糖的酶促释放以及唾液酸酶和 PNGaseF 酶和 MS 采集参数将通过测试其进行验证。我们将在一组匹配的脑脊液和可重复性上使用该测定。来自患有 AD 风险的轻度受损和健康个体的血浆样本 (n=106) 数据以完整形式呈现。聚集蛋白质量峰在血浆和脑脊液之间移动，在唾液酸酶和 PNGaseF 处理、簇蛋白糖型的相对比率以及释放聚糖的 ID 和比率，将在整个队列中进行比较，以及与 apoE 基因型、apoE 异构体特异性糖基化、Aβ42、将检查 Tau 和 pTau。第一个队列的研究结果将通过第二个队列 (n=100) 进行验证。我们的价值主张是一种快速且经济高效的分析方法，用于分析聚集蛋白糖型和聚糖，可用于评估 Clusterin 糖基化在 AD 发病机制中的作用。作为评估 AD 风险的生物标志物，或作为开发改变 AD 风险的治疗方法的目标糖基化。