Microfluidic Chip for Genome Analysis

描述（由申请人提供）：疾病的生物标志物以多种形式存在，如染色体数目或结构畸变、表观遗传因素或基因表达改变、血液成分水平异常或翻译后修饰。通常，准确的诊断和对治疗反应的预测不能依赖于单一的生物标志物，而是几种可能非常不同类型的标志物的组合。这个多学科生物工程项目的目标是开发一种通用的，廉价的技术平台，称为“一次性微流控芯片”（dMFC）。为了支持辅助人类生殖和癌症研究和治疗中的诊断工作，这个经验丰富的研究人员团队寻求开发一种能够将单个细胞固定在指定位置的dMFC（目标1），询问多个遗传基因座以灵敏地检测临床样品中染色体数目和结构畸变（目的2-3）以及基因转录物或肿瘤特异性微小RNA的半定量分析（目的4）。虽然这个R21项目将研究测量多种潜在疾病生物标志物的可行性，包括易位，基因扩增和少量甲状腺和乳腺癌细胞中多达三种肿瘤特异性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，例如辅助人类生殖（ART）中的非侵入性基因表达谱预测卵母细胞能力和胚胎存活。因此，这项可行性研究的成功完成将使我们拥有一个多功能的原型dMFC，以开发更多的临床导向的协议在随后的研究。

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.