Sexually dimorphic miR-497 regulates alpha-synuclein and alpha-synucleinopathy

性二态性 miR-497 调节 α-突触核蛋白和 α-突触核蛋白病

基本信息

批准号：
8638195
负责人：
PETER T. NELSON
金额：
$ 22.5万
依托单位：
UNIVERSITY OF KENTUCKY
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-09-25 至 2015-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8638195
关键词：
Age Age-Years Alzheimer&apos s Disease Area Attenuated Autopsy Biochemical Biochemical Pathway Biological Markers Brain Cells Cessation of life Clinical Cultured Cells Data Data Analyses Data Set Databases Dementia Deposition Diagnostic Disease Elderly Female Gender Gene Dosage Gene Expression Genes Goals Human Knowledge Lentivirus Vector Lewy Bodies Lewy Body Dementia Methods MicroRNAs Molecular Neurodegenerative Disorders Neurons PARK7 gene Pathogenesis Pathology Patients Proteins Public Health Rattus Regulation Regulator Genes Reporting Research Personnel Risk Role Sampling Series Small RNA System Testing Therapeutic Viral Woman age group alpha synuclein alpha synuclein gene base combat data registry disease diagnosis high risk male men neocortical nervous system disorder neurochemistry novel novel diagnostics novel therapeutics public health relevance pup research study sex synucleinopathy trend

项目摘要

DESCRIPTION (provided by applicant): Dementia with Lewy bodies (DLB) is a devastating neurodegenerative disease. Cortical Lewy body pathology (LBP), which is the pathological substrate of DLB, is observed in 10%-30% of brains in most autopsy series of dementia patients. At the molecular level, LBP consists of aberrant deposits of alpha-synuclein (alpha-SN) protein and there is evidence that increased alpha-SN gene dosage is seen in some cases of DLB. The biochemical pathways that contribute to LBP are poorly understood and our overall goal is to help fill this knowledge gap. Men have strongly (~three-fold) increased risk for developing autopsy-confirmed cortical LBP compared to women; this is an observation replicated in many autopsy series. Our overarching hypothesis is that a particular microRNA (miRNA; miR-497) contributes to sexually-dimorphic DLB pathogenesis. In preliminary studies we made the following observations: miR-497 is expressed sexually dimorphically in human brain and in rat primary cultured neurons; miR-497 is enriched in human brains with LBP and in LBP- vulnerable brain areas; and, miR-497 expression in cultured cells alters alpha-SN and DJ-1/PARK7 expression. Our lab has expertise in both LBP and miRNAs which enable us to test the hypothesis that miR- 497 regulates alpha-SN expression and may influence LBP in a sexually dimorphic manner. Specific Aim 1: Test the hypothesis that miR-497 is sexually dimorphically expressed in human brain, and dysregulated in human brains with LBP. Proposed studies will represent the state of the art in miRNA profiling using miRNA microarrays to test for associations between miRNA expression and both LBP and gender status. Preliminary data indicate a systematic tendency for sexually dimorphic miR-497 to be dysregulated in brains with LBP and in human brain areas vulnerable to LBP. Specific Aim 2: Test the hypothesis that sexually dimorphic miR-497 regulates alpha-SN expression. Preliminary data indicate that miR-497 regulates alpha-SN. Primary rat neurons will be transduced with lentiviral vectors expressing miR-497 versus other control miRNAs to test the hypothesis that miR-497 regulates alpha-SN. Further experiments in these cells will query whether the regulation of alpha-SN by miR- 497 is direct or via an intermediate regulation of PARK7/DJ-1 gene. Following viral transduction with miR-497 and control miRNAs, we will determine the full list of miR-497 targets in cultured primary neurons. These mechanistic experiments will aid our ultimate goal of therapies to combat this currently untreatable disease. Public health significance: Understanding how sexually dimorphic miR-497 alters alpha-SN expression to induce sex-dimorphic pathology would be a major novel paradigm in neurochemistry with clinical/translational significance for both diagnostic and therapeutic strategies. The primary cel culture system will offer an experimental paradigm for testing novel methods of attenuating the dimorphic expression of miR-497 that drives alpha-SN expression.

描述（由申请人提供）：Lewy身体（DLB）的痴呆症是一种毁灭性的神经退行性疾病。在大多数尸检系列的痴呆症患者中，在10％-30％的大脑中观察到皮质路易斯体病理学（LBP）是DLB的病理底物。在分子水平上，LBP由α-核蛋白（α-SN）蛋白的异常沉积物组成，并且有证据表明，在某些DLB的情况下，α-SN基因剂量增加。有助于LBP的生化途径知之甚少，我们的总体目标是帮助填补这一知识差距。与女性相比，男性有强烈（〜3倍）增加尸检确认皮质LBP的风险；这是许多尸检系列中复制的观察结果。我们的总体假设是一种特定的microRNA（miRNA; miR-497）有助于性二态DLB发病机理。在初步研究中，我们进行了以下观察结果：miR-497在人脑和大鼠原发性培养神经元中的性二态表达； miR-497富含LBP和LBP脆弱大脑区域的人类大脑；并且，培养细胞中的miR-497表达改变了α-SN和DJ-1/PARK7的表达。我们的实验室在LBP和miRNA中具有专业知识，使我们能够检验Mir-497调节α-SN表达的假设，并可能以性二态性方式影响LBP。具体目的1：检验miR-497在人脑中表达的二态表达的假设，并且在LBP的人类大脑中失调。拟议的研究将使用miRNA微阵列在miRNA分析中代表最新技术，以测试miRNA表达与LBP和性别状态之间的关联。初步数据表明，在LBP的大脑和容易受LBP的人类脑区域中，性二态miR-497的系统趋势是在大脑中失调的趋势。具体目标2：检验性二态miR-497调节α-SN表达的假设。初步数据表明miR-497调节α-SN。原代大鼠神经元将用表达miR-497的慢病毒载体与其他对照miRNA进行转导，以检验miR-497调节α-SN的假设。这些细胞中的进一步实验将查询mir-497对α-SN的调节是直接的还是通过PARK7/DJ-1基因的中间调节。在用miR-497和对照miRNA进行病毒转导后，我们将确定培养的原代神经元中miR-497靶标的完整列表。这些机械实验将有助于我们抗击这种目前不可治疗的疾病的最终疗法。公共卫生的意义：了解性二态miR-497如何改变α-SN表达来诱导性二态病理学将是神经化学的主要新型范式，具有诊断和治疗策略的临床/转化意义。原发性CEL培养系统将提供一个实验范式，用于测试新的方法，用于衰减驱动α-SN表达的miR-497的二态表达。