Molecular Diagnosis Utilizing Cerebrospinal Fluid

描述(由申请人提供)：该研究项目的目标是开发能够从脑脊液(CSF)中提取可重复性的mRNA用于后续基因微阵列分析的技术。在这里提出的一系列研究中，将使用脑脊液创建“分子指纹”，这些患者来自：1)患有继发性神经发育和认知障碍的儿童癌症幸存者，以及2)患有阿尔茨海默病(AD)等与年龄相关的神经退行性疾病的受试者。从脑脊液样本中获取的mRNAs将用于通过扩增反义(ARNA)扩增方法和cDNA阵列技术相结合的分析，以提供数百个基因的广泛、同时表示。设计了两个特定的目的来改进从脑脊液中获取RNA的方法，并将其用作RNA扩增和杂交到本文概述的各种cDNA阵列平台的起始材料。 具体目标1：建立一种从两种不同人群的脑脊液中重复提取和扩增mRNA的新方法：接受化疗的儿童癌症患者和老年痴呆患者。 具体目标2：将从Aim 1扩增的RNA与cDNA微阵列技术相结合，使用脑脊液样本创建表达谱，通过将认知状态与脑脊液特异性分子指纹的rnRNA表达相关联，用于痴呆症的分子诊断和对长期存活的儿童癌症患者的认知能力的跟踪。

DESCRIPTION (provided by applicant):The goal of this research project is to develop technologies that enable reproducible mRNA extraction from cerebrospinal fluid (CSF) for subsequent gene microarray analysis. In the series of studies proposed here, a "molecular fingerprint" will be created using CSF from: 1) pediatric cancer survivors with neurodevelopmental and cognitive impairments secondary to chemotherapy treatment and 2) subjects with age-related neurodegenerative disorders such as Alzheimer's disease (AD). mRNAs harvested from CSF samples will be used for analysis by an amplified antisense (aRNA) amplification methodology combined with cDNA array technology to provide an extensive, concurrent representation of hundreds of genes. Two specific aims are designed to refine a method for harvesting RNA from CSF and using this as a starting material for RNA amplification and hybridization to a variety of cDNA array platforms as outlined herein. Specific aim 1: Develop a novel method for reproducibly extracting and amplifying mRNA from CSF of two different populations of patients: subjects exposed to chemotherapy for the treatment of pediatric cancers and aged individuals with dementia. Specific aim 2: Combine the amplified RNA from Aim 1 with cDNA microarray technology to create expression profiles using CSF samples for molecular diagnosis of dementia and follow up of cognitive performance in long-term survivors of pediatric cancers by correlating cognitive status with rnRNA expression for a CSF-specific molecular fingerprint.