Live Brain Gene Detection in Drug Exposure Subjects after Acupuncture Treatment

描述（由申请人提供）：本项目旨在验证和完善一种新型磁共振成像（MRI）技术，该技术使用靶向磁共振对比探针来检测活体动物因安非他明暴露而导致的内源性基因表达改变。我们将使用C57Black6小鼠的成瘾动物模型。本项目要研究的靶点是转录因子fosB和激活蛋白1 （AP-1）的mRNA， AP-1的表达改变与药物成瘾密切相关。目前大多数分子成像方法检测细胞表面蛋白作为基因表达的标记。我们提出的技术检测细胞内基因表达的产物，如mRNA及其产物，可能具有DNA结合能力在活小鼠的大脑。该方法使用具有反义序列的DNA与目标mRNA或具有一致序列的DNA与AP-1结合。在初步研究中，我们用磁共振造影剂标记这些DNA，并在一项概念验证研究中表明，探针能够在急性安非他明注射后通过MRI报告活体动物c-fos mRNA的升高。我们的目标是扩展和提高该技术的灵敏度，并建立MR信号与细胞内目标的组织学评估之间的相关性。通过实现这些目标，我们将能够将这种新的MR技术作为一种定量工具，用于检测其他基因的内源性基因表达，如环AMP响应元件（CREB）和许多其他与药物成瘾有关的基因表达产物。

DESCRIPTION (provided by applicant): This proposed project aims to validate and refine a novel Magnetic Resonance Imaging (MRI) technique that uses targeting MR contrast probes to detect altered endogenous gene expression resulting from amphetamine exposure in living animals. We will use an animal model of addiction in C57Black6 mice. The targets to be investigated in this project are mRNA of a transcription factor fosB and the activator protein 1 (AP-1) whose altered expression is strongly implicated in drug addiction. Most current molecular imaging methods detect cell surface proteins as markers of gene expression. Our proposed technique detects intracellular products of gene expression, such as mRNA and its products that may have DNA binding ability in the brains of living mice. This method uses DNA with antisense sequence to the target mRNA or DNA with consensus sequence for AP-1 binding. In preliminary studies, we labeled these DNA with MR contrast agent and showed in a proof-of-concept study that the probe is capable of reporting elevation of c-fos mRNA live animals using MRI after acute amphetamine injection. Our goals are to extend and improve the sensitivity of this technique and establish the correlate between MR signal and histological assessment of intracellular targets. By achieving these goals, we will be able to apply this novel MR technique as a quantitative tool for the detection of endogenous gene expression of other genes such as cyclic AMP responsive elements (CREB) and a host of other products of gene expression that are implicated in drug addiction. PUBLIC HEALTH RELEVANCE: In this project, we will extend and refine a novel Magnetic Resonance Imaging (MRI) technique that we have developed to assess the efficacy of acupuncture treatment for substance abuse phenotype by detect altered gene expression in the brains of live animal affected by addictive drugs. The goal of this project is assess and correlate the gene and gene expression product modifications as the result of acupuncture treatment for drug addiction. This application will further our understanding of how acupuncture treatment impacts the progression of drug addiction. Our studies will open new avenues for pharmacotherapeutics by determining whether agents directed against fosB itself, or against any of its many target genes, would be useful in the treatment of diverse types of neuropsychiatric disorders.