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：
10573168
负责人：
Tsuneya Ikezu
金额：
$ 39.13万
依托单位：
MAYO CLINIC JACKSONVILLE
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 2025-02-28
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10573168
关键词：
Acceleration Age Age Months Alzheimer&apos s Disease Alzheimer&apos s disease model Anatomy Animal Model Astrocytes Brain Brain Diseases Brain region Cations Cell model Cells Clinical Complex Cre driver Data Dependovirus Development Disease Endosomes Enzyme-Linked Immunosorbent Assay Genetic Goals Hippocampus In Vitro Incubated Inflammatory LoxP-flanked allele Maps Mediating Microglia Molecular Mus Neurofibrillary Tangles Neuroglia Neurons Pathologic Pathology Pattern Phagocytosis Prevention Process Proteins Purinoceptor Series Signal Pathway Sorting Source Specificity Symptoms System TSG101 gene Tauopathies Testing Therapeutic behavior test behavioral phenotyping cell type comparison control exosome experimental study extracellular vesicles feeding hippocampal pyramidal neuron hyperphosphorylated tau in vivo in vivo Model inhibitor microvesicles mouse model multi-electrode arrays neuropathology neurotoxicity novel pharmacologic prevent protein aggregation tau Proteins tau aggregation tau expression tau mutation trafficking uptake

Acceleration Age Age Months Alzheimer&apos s Disease Alzheimer&apos s disease model Anatomy Animal Model Astrocytes Brain Brain Diseases Brain region Cations Cell model Cells Clinical Complex Cre driver Data Dependovirus Development Disease Endosomes Enzyme-Linked Immunosorbent Assay Genetic Goals Hippocampus In Vitro Incubated Inflammatory LoxP-flanked allele Maps Mediating Microglia Molecular Mus Neurofibrillary Tangles Neuroglia Neurons Pathologic Pathology Pattern Phagocytosis Prevention Process Proteins Purinoceptor Series Signal Pathway Sorting Source Specificity Symptoms System TSG101 gene Tauopathies Testing Therapeutic behavior test behavioral phenotyping cell type comparison control exosome experimental study extracellular vesicles feeding hippocampal pyramidal neuron hyperphosphorylated tau in vivo in vivo Model inhibitor microvesicles mouse model multi-electrode arrays neuropathology neurotoxicity novel pharmacologic prevent protein aggregation tau Proteins tau aggregation tau expression tau mutation trafficking uptake

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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.

由高磷酸化tau蛋白的细胞内聚集体组成的神经纤维缠结是迄今为止大多数与阿尔茨海默氏病临床症状（AD）相关的病理学。新兴证据表明细胞外蔬菜（EV），例如外泌体和微泡，转移病理tau蛋白在细胞作为车辆之间，并在不同的大脑区域传播tau病理学。迫切重要的是找到脑衍生的EV的分子基础，该EV严重调节致病性的转运和吸收神经元细胞之间的tau蛋白和受体神经元中tau蛋白的聚集。当前应用的目的是描述P2RX7（一种嘌呤能受体）对EV-的影响介导的tau繁殖。我们的初步数据表明，抑制GSK的小胶质细胞EV分泌 1482160化合物是P2RX7的特定抑制剂，大大降低了CA1和CA3中的Tau聚集锥体神经元细胞和牙齿颗粒细胞，带有p301s tauopathy动物模型。有趣的是汇聚，减少了外泌体特异性的“运输所需的内体分选络合物”（ESCRT） EV标记，TSG101，在同一海马区域，表明可能来自小胶质细胞的EV运输到海马神经元，可能转移并种子错误折叠的tau并加速蛋白质聚集接受神经元。这就是这些数据表明P2RX7在EV介导的TAU上的调节机制 P2RX7抑制剂在AD或其他tauopathy中的繁殖和阳性。我们假设P2RX7严重调节了小胶质细胞和神经元之间电动汽车的转移海马神经元，因此最喜欢传播错误的tau。我们将验证 GSK1482160在tau传播上，通过使用P2RX7删除在P301S Tau小鼠中概括这些发现和腺相关病毒（AAV）基于TAU传播小鼠模型。在AIM 1中，我们将确定P2RX7在P301S小鼠中的全身缺失的影响。分布将评估海马区域中的EV标记和TAU病理学来自GSK1482160-ADMINDERED P301S小鼠。在AIM 2中，我们将确定P2RX7对分泌的影响以及EV和EV相关的TAU的转移及其在体外的聚集效力。这将确定哪个单元类型特别负责P2RX7调节的Tau扩散到海马神经元中。在AIM 3中，我们将确认在AIM 2中选择的细胞类型，以通过细胞类型将电动汽车导出到海马神经元中 P2RX7或TSG101的特定缺失，外泌体合成分子，并验证P2RX7介导的EV 分泌是使用基于AAV的TAU繁殖小鼠模型的tau传播的。成功的这项研究的完成将增强我们对介导GLIA分泌的分子的理解在体外和体内进行神经元，并确定AD的新靶标。