High Amplification Detection of Genetic Cancer Markers

描述（由申请人提供）：在许多癌症中，诊断时机和最终患者预后与早期诊断密切相关，早期诊断是生存的最大预测因素之一。不幸的是，对于许多相同的癌症，没有简单的检测方法可以轻易地做出早期诊断。尽管大量潜在的生物标志物（包括蛋白质和核酸）存在并已在科学文献中得到确认，但在生物体液中检测这些低水平的标志物（可在早期阶段无创收集）的能力往往被证明是困难或不可能的。为了测试这些生物标志物的预测价值，并在临床环境中实施具有高预测价值的生物标志物的患者测试，迫切需要具有罕见且困难的关键特征组合的分子检测技术，包括敏感性，特异性，稳健性，简单性和低成本。提出的研究试图通过将自由基聚合引发物与指示癌症存在的遗传标记物耦合来开发这样一种技术。通过光引发或化学（即氧化还原）引发的自由基聚合，就其性质而言，是一种扩增事件，其中产生的每个自由基通过大量单体单元聚合，每个单体单元都可以通过各种方法检测到。在此之前，我们已经证明，这种技术能够通过聚合每个初始表面结合分子多达1011个单体，从表面上少至1000个生物标记产生宏观可观察的信号。在这里，我们建议将我们之前在自由基聚合方面的成功转化为一种可行的测试，以确定是否存在某些已知的癌症遗传指标。最终，这样的发展将极大地帮助癌症的诊断、鉴定、治疗和许多与基因异常有关的癌症的存活率。我们的总体假设是，利用自由基聚合作为超高信号放大的手段，将大大提高癌症患者的检测和预后。具体来说，我们的目标是(i)开发和增强氧化还原和光引发自由基聚合方案，用于检测遗传生物标志物的信号扩增，（ii）使用自由基链聚合的信号扩增技术检测单碱基突变，以及（iii）使用自由基链聚合的信号扩增技术检测体内源的单碱基突变。在许多癌症中，诊断时机和患者最终预后密切相关：早期诊断是生存的最大预测因素之一。不幸的是，对于许多相同的癌症，没有简单的检测方法可以轻易地做出早期诊断。这项提议的研究试图开发一种高度敏感、可靠、简便的检测方法，用于检测许多癌症中出现的基因异常，其目标是使早期检测成为可能，远远领先于其他现有技术。

DESCRIPTION (provided by applicant): In numerous cancers, diagnosis timing and ultimate patient prognosis are intimately linked with early diagnosis being one of the greatest predictors of survival. Unfortunately, for many of those same cancers no simple test exists through which an early stage diagnosis is readily made. While a vast body of potential biomarkers, both protein and nucleic acid, exists and is identified in the scientific literature, the ability to detect these markers at the low levels present in biological fluids that can be collected non-invasively at early stages often proves difficult or impossible. To test the predictive value of these biomarkers, and to implement patient testing in a clinical setting for biomarkers that prove to have high predictive value, a pressing need exists for molecular detection technologies that have a rare and difficult combination of crucial characteristics including sensitivity, specificity, robustness, simplicity, and low cost. The proposed research attempts to develop just such a technique by coupling free radical polymerization initiators to genetic markers indicative of cancer's presence. Free radical polymerizations, initiated either by photoinitiation or chemical (i.e., redox) initiation are, by their nature, amplification events wherein each radical that is generated polymerizes through a large number of monomer units, each of which can be detected by a variety of means. Previously, we have shown that this technique is able to produce a macroscopically observable signal from as few as 1000 biological markers on a surface by polymerizing as many as 1011 monomers per initial surface bound molecule. Here, we propose to translate our previous success with free radical polymerizations into a viable test for determining the presence or absence of certain known genetic indicators of the presence of cancer. Ultimately, such a development would dramatically aid in cancer diagnosis, identification, treatment, and survival rates for many cancers linked to genetic anomalies. Our overall hypothesis is that utilization of free radical polymerization as a means for ultrahigh signal amplification will lead to vastly improved detection and outcomes for cancer patients. Specifically, we aim to (i) develop and enhance redox and photoinitiated free radical polymerization schemes for signal amplification in the detection of genetic biomarkers, (ii) detect panels of single base mutations using the signal amplification technique of radical chain polymerization, and (iii) detect panels of single base mutations using the signal amplification technique of radical chain polymerization from an in vivo source.In numerous cancers, diagnosis timing and ultimate patient prognosis are intimately linked: early diagnosis is one of the greatest predictors of survival. Unfortunately, for many of those same cancers no simple test exists through which an early stage diagnosis is readily made. The proposed research attempts to develop a highly sensitive, robust, facile test for genetic anomalies that occur in many cancers, andthe goal is to enable early detection, well in advance of other currently available techniques.