Innovative Tools for the Molecular Diagnosis of Cancer

描述(由申请者提供)：国家癌症研究所呼吁为癌症研究和诊断提供准确、实用和负担得起的新工具。Alderon Biosciences，Inc.建议通过开发一种用于定量研究癌症基因表达的开放式生物传感器平台技术来满足这一需求，目标是取代速度较慢且成本更高的方法。这份关于“癌症分子诊断的创新工具”的SBIR提案的工作假设是，Alderon用于癌症基因表达分析的创新型酶增强电化学系统(ESystem)的一次性传感器阵列和电化学读取器将极大地促进癌症生物标记物的研究。在第一阶段的工作中，已经证明了使用Alderon的生物传感器平台技术进行癌症基因表达定量分析的可行性，并对诱导和非诱导癌细胞中的差异基因表达进行了定量。第一阶段工作取得的主要里程碑是展示了足够的靶标检测灵敏度，足以基于酶增强探针检测小组织样本中即使是很少表达的消息，而不需要靶标放大(在小到104个细胞的样本中，将信使核糖核酸目标量化到每个细胞1-3个拷贝)。在第二阶段，将进一步开发和验证新的eSystem阅读器和ePlates，它们具有传感器面积小的电极，需要非常少量的样品和试剂。第二阶段项目的目标是：1)制造和测试新的eSystem组件(改进的分析、小传感器ePlates和ePlates阅读器)，用于同时量化多个癌症基因靶标的表达水平，而不使用靶扩增步骤；2)生产新型eSystem阅读器和96传感器微板的制造原型；3)改进性能，直到eSystem获得与实时定量RT-PCR分析相同或更好的基因表达分析(速度、样本量、成本、可靠性)。高灵敏度的基因表达测量，不需要现有方法的复杂性或成本，已经被证明可以用新的传感器面积小的微板进行，这种微板需要非常少量的样本和试剂。这项先进的电化学技术将进一步开发用于癌症研究和诊断，然后推广到其他人类保健品。当与疾病相关的基因表达产品在估计5-7年后进入临床使用时，市场可能会增长到每种主要疾病的市场价值可能超过10亿美元/年。我们打算借助成熟的技术和战略合作伙伴关系，在研究和测试领域占据战略份额。

DESCRIPTION (provided by applicant): The National Cancer Institute has called for new tools for cancer research and diagnosis that are accurate, practical and affordable. Alderon Biosciences, Inc. proposes to fill this need by developing an open biosensor platform technology for quantitative studies of cancer gene expression, with the goal of replacing slower and more costly methods. The working hypothesis of this SBIR proposal on "Innovative Tools for the Molecular Diagnosis of Cancer'' is that the disposable sensor arrays and electrochemical readers of Alderon's innovative enzyme-enhanced electrochemical system (eSystem) for cancer gene expression analysis will greatly facilitate studies of cancer biomarkers. Feasibility of using Alderon's biosensor platform technology for quantitative cancer gene expression analysis has been demonstrated in Phase I work and differential gene expression was quantified in induced and non-induced cancer cells. The major milestone achieved in Phase I work was demonstration of target detection sensitivity sufficient for enzyme-enhanced probe-based detection of even rarely expressed messages in small tissue samples without need for target amplification (quantification of mRNA targets down to 1-3 copies per cell in samples as small as 104 cells). In Phase II, new eSystem readers and ePlates with small sensor-area electrodes that require very small volumes of samples and reagents will be further developed and validated. Phase II Project Aims are: 1) make and test new eSystem components (improved assays, small-sensor ePlates and ePlate readers) for simultaneous quantification of levels of expression of multiple cancer gene targets without use of target amplification steps; 2) produce manufacturing prototypes of new eSystem readers and 96-sensor microplates; and 3) refine performance until equivalent or better (speed, sample size, cost, reliability) assays of gene expression are obtained with the eSystem compared to quantitative real-time RT-PCR assays. Highly sensitive gene expression measurements without the complexity or cost of existing methods have been proven possible with new small sensor-area microplates that require very small volumes of samples and reagents. This advanced electrochemical technology will be further developed for cancer research and diagnostics, and then extended to other human health products. When disease-related gene expression products reach clinical use in an estimated 5-7 years, markets will likely grow to exceed a billion dollars/yr for each major disease. We intend to reach a strategic share of the market in research and testing with the aid of proven technology and strategic partnerships.