Microfluidic Chip for Genome Analysis

描述(申请人提供)：疾病的生物标志物以各种形式存在，例如数字或结构染色体异常、表观遗传因素或基因表达改变、血液成分异常水平或翻译后修饰。对治疗反应的准确诊断和预测往往不能依靠单一的生物标记物，而是几种可能非常不同类型的标记物的组合。这一多学科生物工程项目的目标是开发一种多功能、廉价的技术平台，称为“一次性微流控芯片”(DMFC)。为了支持在辅助人类生殖和癌症研究和治疗方面的诊断努力，这个经验丰富的研究团队试图开发一种DMFC，能够将单个细胞固定在指定的位置(目标1)，询问多个遗传位点以灵敏地检测临床样本中的数目和结构染色体异常(目标2-3)，以及对基因转录本或肿瘤特异性microRNAs进行半定量分析(目标4)。虽然这个R21项目将研究在少数甲状腺和乳腺癌细胞中测量多个潜在疾病生物标记物的可行性，包括易位、基因扩增和最多三个肿瘤特异性microRNA的表达，但芯片设计可以很容易地进行修改，以适应不同的应用，例如在辅助人类生殖(ART)中的非侵入性基因表达谱分析，以预测卵母细胞的能力和胚胎存活。因此，这项可行性研究的成功完成将使我们拥有一个通用的DMFC原型，以便在后续的研究中开发出更多面向临床的方案。

DESCRIPTION (provided by applicant): Biomarkers for disease exist in a variety of forms such as numerical or structural chromosome aberrations, epigenetic factors or altered gene expression, abnormal levels of blood constituents or post-translational modifications. Often accurate diagnosis and prediction of response to therapy cannot rely on a single biomarker, but a combination of several, potentially very different types of markers. This multidisciplinary bioengineering project targets the development of a versatile, inexpensive technology platform termed 'disposable microfluidic chip' (dMFC). To support diagnostic efforts in assisted human reproduction and cancer research and therapy, this team of experienced investigators seeks to develop a dMFC capable of immobilizing individual cells in defined positions (Aim 1), interrogating multiple genetic loci for sensitive detection of numerical and structural chromosome aberrations in clinical samples (Aims 2-3) as well as the semi- quantitative analysis of gene transcripts or tumor-specific microRNAs (Aim 4). While this R21 project will investigate the feasibility of measuring multiple potential disease biomarkers including translocations, gene amplifications and expression of up to three tumor-specific microRNAs in small numbers of thyroid and breast cancer cells, the chip design can easily be modified to suit different applications such as noninvasive gene expression profiling in assisted human reproduction (ART) to predict oocyte competence and embryo survival. Thus, the successful completion of this feasibility study will equip us with a versatile prototype dMFC to develop more clinically oriented protocols in subsequent studies.