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Targeting emerging P2RX7 signaling pathways in animal models of Alzheimer's disease

针对阿尔茨海默病动物模型中新兴的 P2RX7 信号通路

基本信息

批准号：
9914594
负责人：
Tsuneya Ikezu
金额：
$ 41.25万
依托单位：
BOSTON UNIVERSITY MEDICAL CAMPUS
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-02-01 至 2020-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9914594
关键词：
Age Age-Months Alzheimer&apos s Disease Alzheimer&apos s disease model Anatomy Animal Model Astrocytes Brain Brain Diseases Brain region Cations Cell Aggregation Cell model Cells Clinical Complex Cre driver Data Dependovirus Development Disease Enzyme-Linked Immunosorbent Assay Genetic Goals Hippocampus (Brain)In Vitro Incubated Inflammatory LoxP-flanked allele Maps Mediating Microglia Molecular Mus Neurofibrillary Tangles Neuroglia Neurons Pathologic Pathology Pattern Phagocytosis Pharmacology Prevention Process Proteins Purinoceptor Seeds Series Signal Pathway Sorting - Cell Movement Source Specificity Symptoms System TSG101 gene Tauopathies Testing Therapeutic base behavior test behavioral phenotyping cell type exosome experimental study extracellular vesicles feeding hyperphosphorylated tau in vivo in vivo Model inhibitor/antagonist microvesicles mouse model multi-electrode arrays neuropathology neurotoxicity novel prevent protein aggregation tau Proteins tau aggregation tau expression tau mutation trafficking uptake

项目摘要

Neurofibrillary tangles, composed of intracellular aggregates of hyperphosphorylated tau protein are by far the most correlated pathology with clinical symptoms of Alzheimer disease (AD). Emerging evidence suggests that extracellular vesicles (EVs), such as exosomes and microvesicles, transfer pathological tau protein between cells as vehicles, and propagate tau pathology in different brain regions. It is urgently important to find the molecular basis of brain-derived EV, which critically regulates the transport and uptake of pathogenic tau protein between neuronal cells and aggregation of tau protein in recipient neurons. The purpose of the current application is to delineate the effect of P2RX7, a purinergic receptor, on EV- mediated tau propagation. Our preliminary data have shown that suppressing microglial EV secretion by GSK 1482160 compound, a specific inhibitor of the P2RX7, dramatically reduces tau aggregation in CA1 and CA3 pyramidal neuronal cells and dentate granular cells with P301S tauopathy animal model. Interestingly this coalesces with reduction of exosome-specific ‘endosomal sorting complexes required for transport’ (ESCRT) EV marker, TSG101, in the same hippocampal regions, suggesting the possible EV trafficking from microglia to hippocampal neurons, which may transfer and seeds misfolded tau and accelerate protein aggregation in receiving neurons. Thus, those data indicate the regulatory mechanism by P2RX7 on EV mediated tau propagation and posit the therapeutic potential of the P2RX7 inhibitor for AD or other tauopathy. We hypothesize that P2RX7 critically regulates the transfer of EVs between microglia and neurons in the hippocampal neurons, thereby facilitate spreading misfolded tau. We will validate the effect of GSK1482160 on tau propagation by recapitulating those findings using P2rx7 deletion in P301S tau mice and adeno-associated virus (AAV)-based tau propagation mouse model. In Aim 1, we will determine the effect of systemic deletion of P2rx7 in P301S mouse. The distribution of EV markers and tau pathology in the hippocampal regions will be evaluated and compared with the findings from GSK1482160-administered P301S mice. In Aim 2, we will determine the effect of P2RX7 on secretion and transfer of EV and EV-associated tau and its aggregation potency in vitro. This will determine which cell type is particularly responsible for P2RX7 regulated tau spread into hippocampal neurons. In Aim 3, we will confirm the cell type, which is selected in Aim 2, for the export of EVs into hippocampal neurons by cell type- specific deletion of P2rx7 or Tsg101, an exosome synthesis molecule, and validate if P2RX7-mediated EV secretion are responsible for tau propagation using AAV-based tau propagation mouse model. Successful completion of this study will enhance our understanding of molecules that mediates secretion of EVs from glia to neurons in vitro and in vivo, and identify novel targets for AD.

由过度磷酸化的tau蛋白的细胞内聚集体组成的神经元缠结是迄今为止最常见的神经元缠结。大多数与阿尔茨海默病（AD）的临床症状相关的病理学。新出现的证据表明细胞外囊泡（EV），例如外来体和微囊泡，转移病理性tau蛋白在细胞之间作为载体，并在不同的大脑区域传播tau病理。当务之急是找到脑源性EV的分子基础，它关键地调节病原体的运输和摄取，神经元细胞之间的tau蛋白和受体神经元中的tau蛋白聚集。本申请的目的是描述嘌呤能受体P2 RX 7对EV-1的作用。介导的tau增殖。我们的初步数据表明GSK抑制小胶质细胞EV分泌 1482160化合物，一种P2 RX 7的特异性抑制剂，显著减少CA 1和CA 3中的tau聚集锥体神经元细胞和齿状颗粒细胞与P301 S tau蛋白病动物模型。有趣的是，合并减少外泌体特异性的“转运所需的内体分选复合物”（ESCRT） EV标志物TSG 101在相同的海马区域，表明EV可能从小胶质细胞运输海马神经元，这可能会转移和种子错误折叠的tau蛋白，并加速蛋白质聚集，接收神经元。因此，这些数据表明P2 RX 7对EV介导的tau的调节机制本发明的目的是提供P2 RX 7抑制剂用于AD或其他tau蛋白病的治疗潜力。我们假设P2 RX 7关键性地调节EV在小胶质细胞和神经元之间的转移，海马神经元，从而促进扩散错误折叠的tau蛋白。我们将验证通过在P301 S tau小鼠中使用P2 rx 7缺失重现这些发现，腺相关病毒（AAV）的tau增殖小鼠模型。在目的1中，我们将确定P2 rx 7系统性缺失在P301 S小鼠中的作用。的分布将评估海马区的EV标志物和tau病理学，并与研究结果进行比较。来自GSK 1482160给药的P301 S小鼠。在目标2中，我们将确定P2 RX 7对分泌的影响。以及EV和EV相关tau的转移及其体外聚集效力。这将决定哪个细胞型特别负责P2 RX 7调节的tau扩散到海马神经元中。在目标3中，我们通过细胞类型确认目标2中选择的用于将EV输出到海马神经元中的细胞类型，特异性缺失P2 rx 7或Tsg 101（一种外泌体合成分子），并验证P2 rx 7介导的EV 使用基于AAV的tau繁殖小鼠模型，分泌的蛋白质负责tau繁殖。成功这项研究的完成将增强我们对介导神经胶质细胞分泌EV的分子的理解神经元在体外和体内，并确定新的目标，为AD。