Brain protein alteration by vascular overexpressed miRNA (BravomiR)

意义：阿尔茨海默病（AD）是最常见的年龄相关性痴呆，影响超过5.8 数百万美国人糖尿病影响着3400万美国人（其中90-95%是2型糖尿病，T2 D），其中 1.5去年新确诊的病例为1000万例。这些“新”病例大多数为45-64岁， 对AD前病因学很重要。大约有8800万人患有糖尿病前期。由于合并症在人类中很常见， AD和糖尿病的综合研究将产生重大的转化影响。我们的目标是揭示特定的共享 AD/T2 D调节通路对于为共病疾病的“联合治疗”奠定基础具有重要意义 原理：了解T2 D-AD轴的共同调节将有利于两个广泛相互关联的 慢性疾病AD神经病理学主要由淀粉样β（Aβ）肽神经炎斑、神经纤维变性、神经纤维变性和神经纤维变性组成。 过度磷酸化tau（τ）的神经缠结、神经炎症和突触丢失。高血压增加 Aβ生成。“通过”T2 D并“进入”AD的其他途径包括脑胰岛素抵抗，以及 糖尿病引发炎症和氧化应激的升高，加剧Aβ和tau聚集。等 共同的稳态失调提示共同的调节障碍。其中一种失调途径就是microRNAs miRNA是一种小的非编码RNA，可调节mRNA翻译成蛋白质。 假设：在T2 D中失调的两种特异性miRNA种类（miR 146 a，miR 200 b）也是 在AD中失调，并且显著的AD相关的直接和下游效应伴随着这种共调节， 失调方法：结合相关的非遗传性T2 D动物模型（人类神经元和人类 神经血管）培养操作和人AD样品（和对照脑）。 目的1：检验T2 D对CNS蛋白水平的破坏是通过以下的血管表达来挽救的假设： miR200 b/miR146 a。我们将测试miR 146 a和200 b对小鼠大脑中蛋白质水平的破坏和拯救。 目的2：验证选择的miRNAs直接与AD相关mRNA的3 '-UTR相互作用的假设。我们 将描述miR 146 a和200 b对AD相关蛋白的作用，并显示miR 200 b和146 a的活性 在小鼠模型中，当用人miR 200 b和/或146 a处理时，在人源细胞中具有对应物。 目的3：测试T2 D和AD类似地破坏人脑中选定的miRNA的假设。我们将 确定miR 200 b和-146在T2 D、AD和T2 D +AD尸检脑样品中被干扰的程度。 影响和未来计划：我们将发现和操纵一个在所有阶段共享的miRNA失调网络 的T2 D/AD，允许早期疾病阶段的监管进展是已知的平行。我们会研究神经- 在miRNA过表达/AD交叉转基因模型中的病理学和在AD Tg模型中诱导的T2 D。我们将 设计实验来跟踪T2 D和AD的共同生化进展，以及T2 D， 进入AD。总体影响是通过血管通路对脑蛋白质改变进行有意识的调节。 通过外周给药过量表达miRNA。

Significance: Alzheimer’s disease (AD) is the most common form of age-related dementia, affecting over 5.8 million Americans. Diabetes affects 34 million Americans (90-95% of which is type 2 diabetes, T2D), of which 1.5 million were newly diagnosed in the previous year. Most of these “new” cases were 45-64 years old, also important to pre-AD etiology. About 88 million have pre-diabetes. Since comorbidity is common in humans, an integrated study of AD and diabetes would have significant translational impact. Our goal to unveil specific shared AD/T2D regulatory pathways is significant to lay groundwork for “co-treatment” of comorbid conditions Rationale: Understanding co-regulation of the T2D-AD axis will be beneficial for two widespread interlinked chronic disorders. AD neuropathology primarily consists of neuritic plaques of amyloid-β (Aβ) peptide, neuro- fibrillary tangles of hyperphosphorylated tau (τ), neuroinflammation, and synaptic loss. Hyperglycemia increases Aβ production. Other pathways that operate “through” T2D and “into” AD include brain insulin resistance, and diabetes-triggered elevation of inflammation and oxidative stress, exacerbating Aβ and tau aggregation. Such shared dyshomeostasis suggests shared dysregulation. One such dysregulatory avenue would be microRNAs (miRNA), small non-coding RNAs that modulate translation of proteins from mRNA. Hypothesis: Two specific miRNA species that are dysregulated in T2D (miR146a, miR200b) are also dysregulated in AD, and significant AD-associated immediate and downstream effects accompany this co- dysregulation. Approach: Combining non-genetic T2D animal models, pertinent (human neuronal and human neurovascular) culture manipulation, and human AD samples (and control brains). Aim 1: Test the hypothesis that T2D disruption of CNS protein levels is rescued by vascular expression of miR200b/miR146a. We will test disruption of protein levels in mouse brains and rescue by miR146a and 200b. Aim 2: Test the hypothesis that selected miRNAs directly interact with 3’-UTRs of AD-associated mRNAs. We will characterize miR146a and 200b effects on AD-related proteins and show that activities of miR200b and 146a in mouse models have counterparts in human-origin cells when treated with human miR200b and/or 146a. Aim 3: Test the hypothesis that T2D and AD similarly disrupt selected miRNAs in human brains. We will determine the extent to which miR200b and -146 are perturbed in T2D, AD, and T2D+AD autopsy brain samples. Impact & Future Plan: We will discover and manipulate a network of shared miRNA dysregulation in all stages of T2D/AD, allow regulatory progression of early disease stages to be known in parallel. We will study neuro- pathology in miRNA overexpression/AD cross transgenic models and induced T2D in AD Tg models. We will design experiments to track shared biochemical progression of T2D and AD, as well as T2D that does not progress into AD. The overall impact is to develop intentional regulation of brain proteins alteration via vascular overexpressed miRNA through peripheral administration.