Functional and Transcriptional Profiling of Monocytes in Alzheimer's Disease

阿尔茨海默病中单核细胞的功能和转录谱

• 批准号: 10370121
• 负责人: Howard L Weiner
• 金额: $ 49.23万
• 依托单位: BRIGHAM AND WOMEN'S 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 2024-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10370121
• 关键词: Abeta clearance; Abeta synthesis; Address; Adjuvant; Affect; Age; Alzheimer' s Disease; Alzheimer' s disease patient; Amyloid; Amyloid beta-Protein; Antigen Presentation; Area; Biological Assay; Brain; CD14 gene; Cells; Characteristics; Chemotaxis; Collaborations; Data; Defect; Dementia; Development; Disease; Dose; Event; Faculty; Gene Expression Profile; Gene Expression Profiling; Genes; Genetic Transcription; Hospitals; Human; Immune; Immune system; Impaired cognition; Impairment; In Vitro; Individual; Interferons; Investigation; Lead; Measures; Memory impairment; Microglia; Monitor; Neuraxis; Neurodegenerative Disorders; Nose; Pathogenesis; Pathway interactions; Patient Schedules; Patients; Peripheral; Peripheral Blood Mononuclear Cell; Phagocytes; Phagocytosis; Phagosomes; Phase; Phenotype; Play; Process; Property; Publishing; Reporting; Research; Role; Sampling; Signal Pathway; T-Lymphocyte; Testing; Time; Woman; abeta accumulation; age related; base; biobank; biomarker identification; design; differential expression; experience; experimental study; immunoregulation; macrophage; member; monocyte; mouse model; multicatalytic endopeptidase complex; response; sex; single-cell RNA sequencing; specific biomarkers; tool; transcriptome; uptake

Alzheimer’s disease (AD) is the most common age-related dementia, for which there is currently no disease modifying therapy. We submit our revised proposal in which we now provide a detailed description of the design, analysis and interpretation of the single-cell (sc)RNAseq pipeline we will follow to comprehensively profile the transcriptome of AD monocytes in collaboration with Dr. Martin Hemberg, a faculty member at our Center with extensive experience in scRNAseq. Also, we describe alternative functional assays we will perform using Protollin-treated AD monocytes that will be isolated from the newly identified clusters of the scRNAseq analysis. Phagocytic innate immune cells, including CNS-resident microglia and infiltrating peripheral monocytes/macrophages, lose their ability to restrict Aβ accumulation and contribute to the disease. It has been shown that AD monocytes have impaired uptake and degradation of Aβ, though investigations in this area are limited and published studies are only at the level of total blood monocyte. We applied single cell RNA-seq to perform unbiased transcriptional analysis of peripheral CD14+ monocytes from two early symptomatic AD patients and two sex, age-matched healthy individuals. We found that monocyte subsets from AD patients acquire unique transcriptional signatures compared to healthy donors. The differentially expressed genes we identified are involved in the interferon, antigen presentation, phagosome and chemotaxis pathways and indicate that AD peripheral monocyte subsets acquire transcriptional signatures that may lead to altered immune properties that contribute to the disease. Furthermore, we have previously shown that Protollin, a proteosome- based adjuvant ameliorates disease in AD mouse models by clearing brain amyloid and in new preliminary data we have found that in vitro treatment of human monocytes with Protollin induces increased soluble Aβ uptake. We hypothesize that peripheral monocytes from AD patients undergo changes at the transcriptional and functional level which impair their homeostatic properties and promotes disease development and that in vitro stimulation of AD monocyte subsets with Protollin can modulate their endogenous AD signature and promote a beneficial functional phenotype. We will address our hypothesis in the following specific aims. Aim 1: Transcriptional and functional profiling of monocyte subsets from AD patients. Aim 2: Modulation of AD monocyte subsets by stimulation with Protollin. Our studies will for the first time define a unique AD transcriptional signature and altered functional characteristics of AD monocytes. We will investigate an immune modulating treatment that could ultimately be tested in patients.

阿尔茨海默氏病（AD）是最常见的与年龄相关的痴呆症，目前尚无疾病 修改疗法。我们提交我们的修订建议，现在我们提供了有关设计的详细说明， 单细胞（SC）RNASEQ管道的分析和解释我们将遵循，以全面介绍 单核细胞的转录组与我们中心的教职员工Martin Hemberg博士合作 Scrnaseq的丰富经验。此外，我们描述了我们将使用的替代功能分析 蛋白质处理的AD单核细胞将从ScRNASEQ分析的新鉴定簇中分离出来。 吞噬先天性免疫小球，包括CNS居民小胶质细胞和浸润的外围 单核细胞/巨噬细胞失去限制Aβ积累的能力并有助于该疾病。它一直 表明单核细胞损害了Aβ的摄取和降解，尽管该领域的研究是 有限和已发表的研究仅在总血单核细胞水平上。我们将单细胞RNA-Seq应用于 对两个早期症状AD的外周三核细胞进行无偏的转录分析 患者和两个性别，年龄匹配的健康个体。我们发现来自AD患者的单核细胞子集 与健康的捐助者相比，获得独特的转录特征。我们表达的基因不同 鉴定出参与干扰素，抗原表现，吞噬体和趋化途径，并表明 该广告周围单核细胞子集获取可能导致免疫变化的转录特征 导致该疾病的特性。此外，我们以前已经表明，蛋白质蛋白是一种蛋白质体 - 通过清除大脑淀粉样蛋白和新的初步数据来调整AD小鼠模型中的疾病 我们发现，在体外用蛋白酶治疗人单核细胞会诱导固体Aβ摄取量增加。 我们假设AD患者的周围单核细胞在转录和 功能水平损害其稳态特性并促进疾病的发展，并在体外 用蛋白蛋白刺激单核细胞亚群可以调节其内源性AD特征并促进A 有益的功能表型。我们将在以下特定目标中解决我们的假设。 AIM 1：AD患者的单核细胞子集的转录和功能分析。 AIM 2：通过用蛋白刺激来调节多核细胞子集。 我们的研究将首次定义独特的AD转录签名和更改功能特征 单核细胞。我们将调查最终可以在患者中测试的免疫调节治疗。