Microglial function of GWAS risk factor BIN1 in Alzheimer's disease pathogenesis and inflammatory signaling

GWAS危险因子BIN1在阿尔茨海默病发病机制和炎症信号传导中的小胶质细胞功能

基本信息

批准号：
10524611
负责人：
Srikant Rangaraju
金额：
$ 225.05万
依托单位：
UNIVERSITY OF SOUTH FLORIDA
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-09-15 至 2025-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10524611
关键词：
Ablation Acute Adaptor Signaling Protein Age Alternative Splicing Alzheimer&apos s Disease Alzheimer&apos s disease risk Amyloid Apolipoprotein E Binding Biochemical Brain Brain Diseases Cerebrum Coupling Data Data Set Disease Disease Progression Disease associated microglia Disease model Endocytosis Enhancers Event Foundations Functional disorder Future Gene Expression Genes Goals Human IRF1 gene Immune signaling Impairment In Vitro Inflammatory Inflammatory Response Injections Interferons Investigation Knockout Mice Label Late Onset Alzheimer Disease Lipopolysaccharides Mass Spectrum Analysis Mediating Membrane Microglia Microscopy Modeling Molecular Molecular Analysis Mus Neurodegenerative Disorders Neurons Pathogenesis Pathogenicity Pathology Pathway interactions Phenotype Play Predisposition Proteins Proteomics Recycling Regulation Reporting Research Risk Factors Role Seeds Signal Pathway Signal Transduction Single Nucleotide Polymorphism Synaptic Vesicles TREM2 gene Tauopathies Testing Time Tissues Transcript Transgenic Organisms Up-Regulation Variant Wild Type Mouse abeta deposition amyloid pathology cell type cerebral amyloidosis conditional knockout genome wide association study in vivo innovation insight large scale data multidisciplinary neuroinflammation neuropathology novel prevent protein expression receptor response risk variant tau Proteins transcriptomics vesicular release β-amyloid burden

项目摘要

PROJECT SUMMARY BIN1, an adaptor protein encoded by the second most common susceptibility GWAS risk factor of late-onset AD, regulates membrane dynamics in the context of endocytosis, membrane remodeling, and synaptic vesicle release. Large-scale expression datasets have reported high-level BIN1 expression in microglia, and AD- associated BIN1 SNPs are thought to alter BIN1 expression through a microglia-specific enhancer. However, the precise functional role(s) of microglial BIN1 in regulating AD pathophysiology has not been investigated systematically. Our central hypothesis is that microglial BIN1 plays an essential role in neuroinflammatory signaling through which BIN1 influences AD pathophysiology. Our preliminary studies show that the loss of Bin1 expression in vitro (cultured microglia) and in vivo (microglia-specific cKO mice) profoundly impairs proinflammatory gene expression and the upregulation of several disease-associated microglia (DAM) genes. Our transcriptomic profiling identified BIN1 as a homeostatic microglial regulator with a non-redundant role in activating proinflammatory response upstream of Apoe, Trem2, and Tyrobp, and upstream of PU.1 and IRF1. BIN1 was predicted to regulate type 1 and 2 interferon responses in microglia in vitro and in vivo. Collectively, these findings offer important insights into microglial BIN1 function, demonstrating its significance in brain inflammatory response. The overall objective of this proposal is to explore BIN1’s role in microglia further, especially in the context of AD pathogenesis, and gain molecular insights. The goal of Aim 1 is to generate 5XFAD:Bin1 cKO mice to elucidate microglial BIN1 function in the modulation of cerebral amyloid burden and amyloid-associated pathophysiology. We will conduct detailed biochemical, molecular, and neuropathological characterization and perform transcriptomics profiling of neuroinflammation and DAM transition to understand BIN1’s role in microglial response to amyloid pathology. Aim 2 studies seek to generate PS19:Bin1 cKO mice to elucidate the involvement of microglial BIN1 function in tau pathophysiology and pathology propagation using detailed neuropathology and comprehensive biochemical, proteomics, and molecular analyses. Aim 3 studies will investigate the mechanistic role of BIN1 as a crucial regulator of early inflammatory signaling events in microglia. We will use unbiased and hypothesis-driven approaches to define the microglial BIN1 interactome and elucidate how BIN1 and its binding partners are reorganized in a context-dependent manner to facilitate immune signaling via key microglial receptors. This timely and unique proposal is highly innovative. Our strategy to use microglia-specific inducible Bin1 cKO mice represents the most direct in vivo approach to rigorously investigate how microglial BIN1 regulates AD pathophysiology and gain insights using comprehensive transcriptomics, proteomics, and interactome characterization. We believe that the successful completion of the proposed research will fill significant gaps in our understanding of BIN1 as a risk factor for LOAD and guide future functional characterizations of molecular pathways and pathogenic mechanisms regulated by this major LOAD risk gene.

项目摘要 BIN1，一种适配蛋白，由第二个最常见的敏感性GWAS危险因素编码，在内吞作用，膜重塑和突触囊泡的背景下调节膜动力学发布。大规模表达数据集报道了小胶质细胞中的高级BIN1表达，并且相关的BIN1 SNP被认为可以通过小胶质细胞特异性增强剂改变BIN1的表达。然而，尚未研究小胶质细胞BIN1在调节AD病理生理学中的精确功能作用系统地。我们的中心假设是小胶质细胞BIN1在神经炎症中起着至关重要的作用 BIN1影响AD病理生理学的信号传导。我们的初步研究表明，BIN1的丧失体外（培养的小胶质细胞）和体内（小胶质细胞特异性CKO小鼠）的表达严重损害促炎基因表达和几种疾病相关的小胶质细胞（DAM）基因的上调。我们在激活ApoE，Trem2和Tyrobp上游的促炎反应，以及PU.1和IRF1的上游。预测BIN1在体外和体内调节小胶质细胞中的1型干扰素反应。共同这些发现为小胶质细胞BIN1功能提供了重要的见解，证明了它在大脑中的重要性炎症反应。该提案的总体目的是进一步探索BIN1在小胶质细胞中的作用，特别是在AD发病机理的背景下，并获得分子见解。目标1的目标是产生 5xFAD：BIN1 CKO小鼠在调节大脑淀粉样burnen和淀粉样蛋白相关的病理生理学。我们将进行详细的生化，分子和神经病理学表征和执行神经炎症和大坝过渡的转录组学分析 BIN1在小胶质病对淀粉样病理学的小胶质反应中的作用。 AIM 2研究试图生成PS19：BIN1 CKO小鼠到使用小胶质细胞BIN1功能参与使用Tau病理生理学和病理传播的参与详细的神经病理学和全面的生化，蛋白质组学和分子分析。 AIM 3研究将研究BIN1作为早期炎症信号事件的关键调节剂的机械作用小胶质细胞。我们将使用公正和假设驱动的方法来定义小胶质的BIN1 Itspettootome和阐明BIN1及其结合伙伴如何以上下文依赖性方式重组以促进免疫通过关键小胶质接收器发出信号。这个及时和独特的提议具有很高的创新性。我们使用的策略小胶质细胞特异性诱导BIN1 CKO小鼠代表了严格研究的最直接的体内方法小胶质细胞BIN1如何调节AD病理生理学并使用综合转录组学获得见解，蛋白质组学和相互作用的表征。我们认为，拟议的成功完成研究将填补我们对BIN1作为负载危险因素的理解并指导未来功能的显着空白该主要负荷风险基因调节的分子途径和致病机制的表征。