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.

阿尔茨海默病（AD）是最常见的年龄相关性痴呆，目前没有疾病 改良疗法我们提交了修改后的提案，其中我们现在提供了设计的详细说明， 分析和解释单细胞（sc）RNAseq管道，我们将遵循全面概况 AD单核细胞的转录组与我们中心的教师Martin Hemberg博士合作， 在scRNAseq方面的丰富经验。此外，我们描述了我们将使用 Protollin处理的AD单核细胞，其将从scRNAseq分析的新鉴定的簇中分离。 吞噬性先天免疫细胞，包括CNS驻留的小胶质细胞和浸润性外周 单核细胞/巨噬细胞，失去了限制Aβ积聚的能力，并导致疾病。已经 表明AD单核细胞对Aβ的摄取和降解受损，尽管这方面的研究还不够深入， 有限的和已发表的研究仅在总血液单核细胞水平上。我们将单细胞RNA-seq应用于 对来自两个早期症状性AD的外周CD 14+单核细胞进行无偏转录分析 患者和两个性别、年龄匹配的健康个体。我们发现AD患者的单核细胞亚群 获得独特的转录签名相比，健康的捐助者。差异表达的基因， 已确定参与干扰素、抗原呈递、吞噬体和趋化性途径，并表明 AD外周血单核细胞亚群获得转录特征，可能导致免疫改变， 导致疾病的特性。此外，我们以前已经表明，Protollin，一个蛋白体- 基于佐剂通过清除脑淀粉样蛋白和新的初步数据改善AD小鼠模型的疾病 我们发现用Protollin体外处理人单核细胞可诱导可溶性Aβ摄入增加。 我们假设AD患者外周血单核细胞在转录水平发生变化， 功能水平，损害他们的稳态特性，促进疾病的发展和体外 用Protollin刺激AD单核细胞亚群可以调节其内源性AD特征并促进AD细胞增殖。 有益的功能表型。我们将在以下具体目标中阐述我们的假设。 目的1：AD患者单核细胞亚群的转录和功能分析。 目的2：通过用Protollin刺激来调节AD单核细胞亚群。 我们的研究将首次确定一个独特的AD转录签名和改变的功能特征 AD单核细胞我们将研究一种免疫调节治疗方法，最终可以在患者中进行测试。