Probing Heterogeneity of Alzheimer's Disease Using iPSCs

使用 iPSC 探索阿尔茨海默病的异质性

基本信息

批准号：
10657140
负责人：
Tracy L YOUNG-PEARSE
金额：
$ 85.52万
依托单位：
BRIGHAM AND WOMEN'S HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-07-15 至 2028-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10657140
关键词：
Address Adult Alzheimer&apos s Disease Alzheimer&apos s disease brain Alzheimer&apos s disease risk Amyloid beta-Protein Astrocytes Biology Brain CASP1 gene CD36 gene CRISPR/Cas technology Cells Coculture Techniques Cognitive Constitution Constitutional Data Detergents Development Disease Disease Progression Environment Enzyme-Linked Immunosorbent Assay Exhibits Experimental Models Formic Acids Functional disorder Genes Genetic Transcription Genetic study Genome engineering Health Heterogeneity Heterozygote Human IL18 gene Immune Immune signaling Impairment Individual Induced pluripotent stem cell derived neurons Inflammasome Inflammation Inflammatory Interleukin-1 beta Interleukin-13 Knockout Mice Late Onset Alzheimer Disease Length Link Literature Lysosomes Measurement Medial Mediating Membrane Microglia Modeling Molecular Nerve Degeneration Neurodegenerative Disorders Neurofibrillary Tangles Neuroimmunomodulation Neurons Pathogenesis Pathway interactions Phagocytosis Phosphoric Monoester Hydrolases Play Prefrontal Cortex Process Proteins Proteomics RNA Receptor Up-Regulation Regulation Reporting Risk Role Sampling Senile Plaques Series Signal Pathway Signal Transduction Stains Synapses System TNF gene Transgenic Mice Up-Regulation Variant Visualization Water Western Blotting Work abeta accumulation brain cell brain health brain tissue cell type cohort conditional knockout cytokine experimental study genetic risk factor genome wide association study human model induced pluripotent stem cell inhibitor inorganic phosphate insight mouse model neuroinflammation pharmacologic phosphoinositide-3,4,5-triphosphate phosphoinositide-3,4-bisphosphate receptor upregulation response scavenger receptor single nucleus RNA-sequencing tau Proteins tau-1 transcriptome sequencing

项目摘要

PROJECT SUMMARY There is an increasing focus on understanding the role of microglia in neurodegenerative disorders including late-onset Alzheimer’s disease (LOAD), a disease defined by the accumulation of amyloid beta rich plaques and neurofibrillary tangles containing tau. Microglia are proposed to play a variety of critical roles during disease progression including synaptic engulfment, cytokine release, and phagocytosis of amyloid beta (Aβ). Further, recent GWAS studies have implicated innate immune processes in LOAD, supporting the importance of understanding the neuroimmunological processes underlying the risk and progression of AD. All recent large- scale GWAS have identified SNPs at the INPP5D locus that are significantly associated with AD. INPP5D expression is largely restricted to microglial cells in the human adult brain and we have confirmed previous findings that RNA levels of INPP5D are elevated in AD brain. However, through quantitative western blotting, we show that protein levels of full length, water-soluble INPP5D are reduced in AD brain. To study the functional consequences of reduced INPP5D, we used both pharmacological inhibition and CRISPR-Cas9 genome engineering to lower INPP5D activity in human iPSC-derived microglia. Through unbiased RNA and proteomic profiling and a series of pharmacological manipulations, we demonstrated that reduction of INPP5D activity induces changes in immune signaling and, more specifically, the activation of the inflammasome. These studies have raised important questions that we aim to address in this proposal regarding the constitution of INPP5D in microglia in the Alzheimer’s brain (aim 1), the molecular mechanism(s) linking INPP5D and inflammasome activation (aim 2), and the functional consequences of loss of INPP5D activity and inflammasome activation in microglia on astrocytes and neurons (aim 3). In aim 1, we will utilize quantitative immunostaining, sequential extraction and western blotting, and ELISA to deeply interrogate INPP5D levels and markers of inflammasome activation across a large cohort of human brain samples and iPSC-derived microglia cultures. Aim 2 interrogates candidate pathways that may link INPP5D activity with inflammasome activation that arose from our preliminary analyses. The first of these involves upregulation of PLA2G7, second through dysregulation of scavenger receptors and the third through disruption of lysosome function. Finally, in aim 3 we utilize co-culture models of iPSC-derived neurons, astrocytes and microglia and a conditional INPP5D knock out mouse model to determine the consequences of reduction of INPP5D levels in microglia on neuron and astrocyte biology in the context of AD relevant environments. Together, these studies will provide important new insights into our understanding of the fundamental role of INPP5D in microglia in health and disease, regulation of inflammasome activation in human microglia, and the consequences of sub-lytic inflammasome activation in microglia on neuron and astrocyte biology.

项目摘要越来越关注理解小胶质细胞在神经退行性疾病中的作用，包括晚发的阿尔茨海默氏病（负载），这是一种由淀粉样蛋白β丰富斑块的积累定义的疾病含有tau的神经纤维缠结。提议小胶质细胞在疾病中起多种关键作用进展包括突触吞噬，细胞因子释放和淀粉样β（Aβ）的吞噬作用。更远， GWAS最近的研究已经实施了载荷的先天免疫程序，支持了了解AD的风险和进展为基础的神经免疫学过程。所有最近的大型比例GWA已鉴定出与AD显着相关的INPP5D基因座的SNP。 inpp5d 表达在很大程度上仅限于人类大脑中的小胶质细胞，我们先前已经证实发现AD大脑中INPP5D的RNA水平升高。但是，通过定量的蛋白质印迹，我们表明在AD大脑中，全长，水溶性INPP5D的蛋白质水平降低。研究功能降低INPP5D的后果，我们同时使用了药物抑制和CRISPR-CAS9基因组在人IPSC衍生的小胶质细胞中降低INPP5D活性的工程。通过无偏的RNA和蛋白质组学分析和一系列药物操作，我们证明了INPP5D活性的降低诱导免疫信号传导的变化，更具体地说是炎症体的激活。这些研究已经提出了我们旨在在此提案中解决有关Inpp5d配置的重要问题阿尔茨海默氏症大脑中的小胶质细胞（AIM 1），分子机制与INPP5D和炎症体连接激活（AIM 2），以及INPP5D活性丧失和炎症体激活的功能后果星形胶质细胞和神经元的小胶质细胞（AIM 3）。在AIM 1中，我们将利用定量免疫染色，顺序提取和蛋白质印迹，以及ELISA，深入询问INPP5D水平和炎性体标志物大量人脑样品和IPSC衍生的小胶质细胞培养物的激活。目标2询问可能将INPP5D活性与炎症体激活联系起来的候选途径，这是我们的初步引起的分析。其中的第一个涉及PLA2G7的上调，其次是通过清除剂失调受体和第三次通过溶酶体功能的破坏。最后，在AIM 3中，我们利用了共同文化模型 IPSC衍生的神经元，星形胶质细胞和小胶质细胞以及有条件的INPP5D敲除小鼠模型在神经元和星形胶质细胞生物学的背景下，小胶质细胞中INPP5D水平降低的后果广告相关环境。这些研究将共同为我们的理解提供重要的新见解 INPP5D在小胶质细胞在健康和疾病中的基本作用，调节炎症体激活人类小胶质细胞，以及小胶质细胞中的亚乳化炎性体在神经元和星形胶质细胞生物学。

项目成果

期刊论文数量（10）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

RNA-binding protein ELAVL4/HuD ameliorates Alzheimer's disease-related molecular changes in human iPSC-derived neurons.

DOI：
10.1016/j.pneurobio.2022.102316
发表时间：
2022-10
期刊：
PROGRESS IN NEUROBIOLOGY
影响因子：
6.7
作者：
van der Linden, Robert J.;Gerritsen, Jacqueline S.;Liao, Meichen;Widomska, Joanna;Pearse II, Richard V.;White, Forest M.;Franke, Barbara;Young-Pearse, Tracy L.;Poelmans, Geert
通讯作者：
Poelmans, Geert

Loss of endosomal exchanger NHE6 leads to pathological changes in tau in human neurons.

DOI：
10.1016/j.stemcr.2022.08.001
发表时间：
2022-09-13
期刊：
STEM CELL REPORTS
影响因子：
5.9
作者：
Fernandez, Marty A.;Bah, Fatmata;Ma, Li;Lee, YouJin;Schmidt, Michael;Welch, Elizabeth;Morrow, Eric M.;Young-Pearse, Tracy L.
通讯作者：
Young-Pearse, Tracy L.

Lost in translational biology: Understanding sex differences to inform studies of diseases of the nervous system.

迷失在转化生物学中：了解性别差异为神经系统疾病的研究提供信息。

DOI：
10.1016/j.brainres.2019.146352
发表时间：
2019
期刊：
Brain research
影响因子：
2.9
作者：
Pearse2nd,RichardV;Young-Pearse,TracyL
通讯作者：
Young-Pearse,TracyL

Mosaic loss of Chromosome Y in aged human microglia.

DOI：
10.1101/gr.276409.121
发表时间：
2022-10
期刊：
Genome research
影响因子：
7
作者：
Vermeulen MC;Pearse R;Young-Pearse T;Mostafavi S
通讯作者：
Mostafavi S

Small Molecule Regulators of microRNAs Identified by High-Throughput Screen Coupled with High-Throughput Sequencing.

通过高通量筛选结合高通量测序鉴定 microRNA 的小分子调节因子。

DOI：
10.21203/rs.3.rs-2617979/v1
发表时间：
2023
期刊：
Research square
影响因子：
0
作者：
Krichevsky,Anna;Nguyen,Lien;Wei,Zhiyun;Silva,M;Barberán-Soler,Sergio;Rabinovsky,Rosalia;Muratore,Christina;Stricker,Jonathan;Hortman,Colin;Young-Pearse,Tracy;Haggarty,Stephen
通讯作者：
Haggarty,Stephen