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。在本文提出的一系列研究中，将使用CSF创建“分子指纹”：1）患有继发于化疗治疗的神经发育和认知障碍的儿童癌症幸存者和2）患有年龄相关神经退行性疾病（如阿尔茨海默病（AD））的受试者。从CSF样本中收获的mRNA将用于通过扩增反义（aRNA）扩增方法结合cDNA阵列技术进行分析，以提供数百种基因的广泛、同时代表性。设计了两个具体目的以改进从CSF收获RNA并将其用作RNA扩增和杂交到如本文所述的多种cDNA阵列平台的起始材料的方法。 具体目标1：开发一种新的方法，用于从两种不同患者人群的CSF中可重复地提取和扩增mRNA：暴露于用于治疗儿科癌症的化疗的受试者和患有痴呆症的老年人。 具体目标2：联合收割机将来自Aim 1的扩增RNA与cDNA微阵列技术相结合，使用CSF样本创建表达谱，通过将认知状态与mRNA表达相关联以获得CSF特异性分子指纹，用于痴呆的分子诊断和儿科癌症长期存活者的认知表现随访。

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.