Neurobiological Role of MicroRNA in Alzheimer's

我们的目标是确定导致阿尔茨海默病(AD)神经退行性变的机制。我们会研究 淀粉样蛋白β前体蛋白、微管相关蛋白tau、−突触核蛋白的调控 (SNCA)和Re1-沉默转录因子(REST)。这项提案将考验miR153作为 监管者的。我们已经确定miR153调节重要的AD相关mRNAs的翻译 APP、SNCA和REST通过非翻译区(UTR)。我们建议检验miR153的假设 作为多种AD相关蛋白的神经网络调节器，控制神经元存活。这项提议将 显著促进了对miR153‘S在调节神经退行性变相关蛋白中的作用的理解。 SA1.分析miR153与REST-153网络(R-153net)伙伴mRNAs中的目标站点相互作用。这个 REST、APP、tau、IL-1、α和IL-6蛋白在AD中起重要作用。控制这种蛋白质的“主调节器” 血药浓度可能是预防AD相关神经毒性的重要工具。我们将测试miR153在多个 基于选定基因的信使核糖核酸序列的融合报告克隆。我们期待miR153治疗 将沉默预测的靶点，而miRNA治疗将进一步改变细胞培养中的蛋白质水平。 SA2.测试miR153干扰R-153net的表达和细胞的健康和生长。我们将定义 MiR153‘S相互作用的神经生物学，包括活性氧化物种(ROS)。我们将在以下方面确立效力 天然蛋白质水平和细胞存活率。确定miRNA介导的R-153net Will的调节作用 阐明miRNA依赖的增强细胞活力的机制。我们预计miR153与总体 这些研究将揭示miR153在细胞生存中的作用机制。 SA3.测量miR153治疗改变诱导多能干细胞(IPSC)中的R-153net。我们会 检测miR153对正常和AD供者分化的IPSC的作用。我们预计MIR153 将改变R-153net水平以及细胞活力和形态。将IPSC培养成神经元、神经胶质细胞和 混合类型细胞诱导将允许明确地测量每一种主要脑细胞类型的影响。 SA4.评估miR153在AD分期和脑区特异性方式中表达异常。我们将测试和 模型miR153在AD、MCI和对照人脑样本中的mRNA和蛋白质水平，以及APOE， 性别、年龄与疾病进展的关系。我们预计miR153和R-153net蛋白质和mRNAs的水平会有所不同 以依赖于进程的方式。我们的结果将预测与R-153net成员级别相关的风险。 SA5.确定MIR153基因附近的SNP与改变的脑脊液A、β和磷酸化tau水平相关。我们 将确定接近MIR153-1和-2基因的SNPs的影响。我们将研究大规模的基因组 SNP与AD内表型的相关性数据，这将提供预测性AD生物标记物。我们的研究 在细胞培养中，ipscs，miRNA在人脑中的表达，并识别MIR153基因附近的SNPs 将建立它们与阿尔茨海默病的关系，并导致新的诊断和治疗策略。

Our goal is to identify mechanisms leading to neurodegeneration in Alzheimer's disease (AD). We will study the regulation of amyloid-β precursor protein (APP), microtubule-associated protein tau (MAPT), −Synuclein (SNCA), and RE1-Silencing Transcription Factor (REST). This proposal will test miR153 as a 'regulator of the regulators'. We have established that miR153 regulates the translation of important AD-associated mRNAs for APP, SNCA, and REST via the untranslated region (UTR). We propose testing the hypothesis that miR153 serves as a nexus regulator of multiple AD-related proteins and control neuronal survival. This proposal will significantly advance the understanding of miR153's role in regulating proteins related to neurodegeneration. SA1. Analyze miR153 interacts with target sites in REST-153 network (R-153net) partner mRNAs. The proteins REST, APP, tau, IL1α, and IL6 play critical roles in AD. A "master regulator" that controls such protein levels could be a vital tool to prevent AD-related neurotoxicity. We will test the activity of miR153 on multiple mRNA-fused reporter clones based on the mRNA sequence of selected genes. We expect miR153 treatments will silence predicted target sites, and miRNA treatment will further alter protein levels in cell cultures. SA2. Test miR153 perturbs expression of the R-153net and cell health and growth. We will define the neurobiology of miR153's interactions, including reactive oxidizing species (ROS). We will establish effects on native protein levels and cell survival. Identifying the roles of miRNA-mediated regulation of the R-153net will elucidate miRNA-dependent mechanisms to enhance cell vitality. We expect that miR153 interacts with overall cell health, and these studies will reveal mechanisms for miR153 function in cell survival. SA3. Measure miR153 treatment alters the R-153net in induced pluripotent stem cells (iPSC). We will test the effects of miR153 treatment in differentiated iPSC from normal and AD donors. We expect that miR153 will alter R-153net levels and cell vitality and morphology. "Forking" iPSC cultures into neurons, glia, and mixed-type cell induction will allow for explicitly measuring effects in each of the major brain cell types. SA4. Assess miR153 is dysregulated in AD stage and brain region-specific manners. We will test and model miR153, mRNA, and protein levels in AD, MCI, and control human brain samples, along with APOE, sex, and age vs. disease progression. We expect levels of miR153 and R-153net proteins and mRNAs to vary in progression-dependent manners. Our results will predict the risk associated with R-153net member levels. SA5. Identify SNPs near MIR153 genes associate with altered CSF Aβ and phospho-tau levels. We will identify the effects of SNPs close to the MIR153-1 and -2 genes. We will investigate large-scale genomic data for SNP associations with AD endophenotypes, which will provide predictive AD biomarkers. Our studies in cell cultures, iPSCs, miRNA expression in human brains, and identifying SNPs near MIR153 genes will establish their relationships to AD and lead to novel diagnostic and therapeutic strategies.