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，没有简单的测试存在，通过它的早期诊断是容易的。虽然存在大量潜在的生物标志物（蛋白质和核酸）并且在科学文献中鉴定，但是在可以在早期阶段非侵入性地收集的生物流体中以低水平检测这些标志物的能力通常被证明是困难的或不可能的。为了测试这些生物标志物的预测价值，并且为了在临床环境中对证明具有高预测价值的生物标志物实施患者测试，迫切需要具有关键特征（包括灵敏度、特异性、稳健性、简单性和低成本）的罕见且困难的组合的分子检测技术。这项拟议中的研究试图通过将自由基聚合引发剂与指示癌症存在的遗传标记结合来开发这样一种技术。通过光引发或化学引发的自由基聚合（即，氧化还原）引发就其性质而言是放大事件，其中产生的每个自由基通过大量单体单元聚合，每个单体单元可以通过多种手段检测。以前，我们已经表明，这种技术是能够产生一个宏观上可观察到的信号，从少至1000个生物标记物的表面上聚合多达1011个单体每个初始表面结合分子。在这里，我们建议将我们以前的自由基聚合成功转化为一个可行的测试，用于确定是否存在某些已知的癌症存在的遗传指标。最终，这样的发展将大大有助于癌症的诊断，识别，治疗和与遗传异常有关的许多癌症的生存率。我们的总体假设是，利用自由基聚合作为一种手段，用于信号放大将导致大大改善癌症患者的检测和结果。具体而言，我们的目标是（i）开发和增强氧化还原和光引发的自由基聚合方案，用于检测遗传生物标志物中的信号放大，（ii）使用自由基链聚合的信号放大技术检测单碱基突变组，以及（iii）使用自由基链聚合的信号放大技术从体内来源检测单碱基突变组。诊断时机和最终患者预后密切相关：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，没有简单的测试存在，通过它的早期诊断是容易的。这项拟议中的研究试图开发一种高度敏感、稳健、简便的检测方法，用于检测许多癌症中发生的遗传异常，其目标是在其他现有技术之前进行早期检测。

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.