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Imaging the metabolic and phagocytic landscape of microglia in Alzheimer’s disease

对阿尔茨海默病中小胶质细胞的代谢和吞噬景观进行成像

基本信息

批准号：
10393001
负责人：
Sarah C Heilshorn
金额：
$ 15.74万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-04-15 至 2023-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10393001
关键词：
Alzheimer&apos s Disease Alzheimer&apos s disease brain Alzheimer&apos s disease diagnosis Alzheimer&apos s disease pathology Alzheimer&apos s disease risk Amyloid Amyloid beta-Protein Brain Cells Characteristics Chronic Clustered Regularly Interspaced Short Palindromic Repeats Complement Data Deposition Disease Disease Progression Disease associated microglia Environment Exposure to Fluorescence Microscopy Gene Expression Profile Genes Genetic Genetic Risk Genetic Transcription Health Heterogeneity Human Human Cell Line Human Genome Image Immune In Situ Hybridization Individual Inflammatory Information Distribution Intervention Knock-out Link Lipids Lipofuscin Maps Messenger RNA Metabolic Microglia Microscope Microscopy Mitochondria Molecular Myelin Nerve Degeneration Neurites Oxidation-Reduction Pathologic Pathology Persons Phagocytes Phenotype Play Proteins RNA RNA Probes Role Route Tissues Transcript base brain cell brain parenchyma brain tissue cytokine disorder prevention fluorophore gene discovery genome wide association study genome-wide imaging probe lipid metabolism metabolic imaging metabolic phenotype metabolic profile microscopic imaging new technology new therapeutic target novel therapeutics protein aggregation reduce symptoms response risk variant single cell analysis single-cell RNA sequencing symptom treatment therapeutic target trait two-photon wasting

项目摘要

Imaging the Metabolic and Phagocytic Landscape of Microglia in Alzheimer’s Disease Genome-wide association studies show that some of the strongest genetic risk variants for Alzheimer’s disease (AD) involve genes exclusively expressed in microglia, indicating its central role in AD pathology. Microglia are the resident immune cells of the brain, essential for maintaining the health and function of the brain, as well as providing a first line of defense by phagocytizing debris and secreting cytokines. In AD, characterized by a CNS environment with chronic exposure to cellular debris and protein aggregation, recent single-cell RNA sequencing has uncovered a variety of microglial transcriptional states specific to AD, indicating both protective and detrimental functions. While their transcriptional profiles are well-characterized, we lack an understanding of the molecular mechanisms that drive the formation of protective/detrimental microglial phenotypes, their functional characteristics and how they inform AD disease pathology. Only by connecting transcriptional profiles to functional cell-states can we identify promising, new therapeutic targets. Subpopulations with detrimental functional signatures may represent novel therapeutic targets for AD. This requires the integration of new technologies into the field, where the transcriptional profile of individual cells can be complemented by their functional signatures and correlated with microglia-activating agents and pathological hallmarks. Here, we propose to complement available transcriptional data with microscopy of the metabolic and phagocytic landscape of microglia in the human AD brain. The heterogeneity of microglial transcription makes it difficult to unambiguously distinguish phenotypes based on immunostaining in conventional fluorescence microscopy. Instead, we have developed a front-line nonlinear microscopy platform, where microglial phenotypes can be distinguished based on their metabolic and phagocytic profiles using spectral coherent anti-Stokes Raman (CARS) and simultaneous two-photon excited fluorescence (TPEF) microscopy. The profiles will be compiled from quantitative data extracted from the microscopy images; amounts of (i) intracellular lipid stores, (ii) mitochondria, and (iii) the cellular redox ratio will be integrated into the metabolic profile, while (iv) lysosomal myelin/amyloid debris, (v) cytosolic myelin debris, and (vi) accumulating undegradable waste as lipofuscin will form the phagocytic profile. By further integrating a capability to map the distribution of specific RNA transcripts using RNA probes (RNAScope), we will be able to link transcriptional expression to the metabolic and phagocytic profiles at the single cell level. Specifically, we will investigate the metabolic and phagocytic signatures of microglia in human AD brain tissues that express a set of genes, which we have discovered to modulate lipid accumulation in human immune cells through our functional genome-wide CRISPR knock-out screens. This will reveal genetic regulators of dysfunctional lipid accumulation, characteristic for detrimental microglia, which may represent novel therapeutic targets. We envision that metabolic reprogramming of microglia will become a new therapeutic route for AD.

阿尔茨海默病小胶质细胞代谢和吞噬的影像学研究