Molecular Diagnosis Utilizing Cerebrospinal Fluid

描述（由申请人提供）：本研究项目的目标是开发能够从脑脊液（CSF）中提取可重复mRNA的技术，用于随后的基因微阵列分析。在这里提出的一系列研究中，将利用脑脊液创建一个“分子指纹”：1)化疗后继发神经发育和认知障碍的儿科癌症幸存者，2)与年龄相关的神经退行性疾病（如阿尔茨海默病（AD））受试者。从脑脊液样品中收集的mrna将通过扩增反义（aRNA）扩增方法与cDNA阵列技术相结合进行分析，以提供数百个基因的广泛，并发代表。设计了两个特定的目标，以改进从CSF中收集RNA的方法，并将其作为RNA扩增和杂交到各种cDNA阵列平台的起始材料。

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.