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) 中重复提取 mRNA 的技术，用于后续基因微阵列分析。在此提出的一系列研究中，将使用脑脊液创建“分子指纹”：1）因化疗治疗而继发神经发育和认知障碍的儿科癌症幸存者，2）患有与年龄相关的神经退行性疾病（例如阿尔茨海默氏病（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，以提供数百个基因的广泛、同时表示。设计了两个具体目标来改进从 CSF 中收获 RNA 的方法，并将其用作 RNA 扩增和与本文概述的各种 cDNA 阵列平台杂交的起始材料。 具体目标 1：开发一种新方法，用于从两个不同患者群体的脑脊液中可重复地提取和扩增 mRNA：接受儿科癌症化疗的受试者和患有痴呆症的老年个体。 具体目标 2：将目标 1 中扩增的 RNA 与 cDNA 微阵列技术相结合，利用 CSF 样本创建表达谱，用于痴呆症的分子诊断，并通过将认知状态与 CSF 特异性分子指纹的 RNRNA 表达相关联来跟踪儿科癌症长期幸存者的认知表现。

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.