Development of a blood test for breast cancer detection

描述(申请人提供)：乳房X光检查是乳房疾病检测和诊断的基准。然而，它可能会漏掉10%-15%的早期乳腺癌，并且无法确定地区分良恶性病变。提供一种替代方法，例如基于生物标记物的方法，使良性疾病和乳腺癌之间能够及早发现/精确区分，这将降低与乳腺癌相关的死亡率。我们之前使用DNA微阵列筛选了&GT；300个血液RNA样本，并识别了24个RNA签名，这些签名允许精确识别选定的乳腺疾病患者。基于已识别的生物标记物作为基于血液的乳腺疾病检测检测的商业化前景，我们建议使用更易于转化为临床的平台来验证候选生物标记物。目的：建立一种基于生物标志物的血液检测方法，用于乳腺疾病的检测和分类。该项目的具体目标是：1)设计和构建用于多重血液RNA分析的Quantigene探针，2)用血液RNA样本测试探针，并开发用于识别良性乳腺疾病(BD)和乳腺癌(BC)个体的原型分类模型，3)验证原型模型在识别捐赠者类别方面的性能。方法：设计和构建针对24个感兴趣的生物标志物和3个看家基因的Quantigene RNA探针。血液采集采用PAX基因RNA稳定管，采自正常(n=30)、BD(n=30)和BC(n=30)的女性献血者(&gt；21岁)，并用Quantigene探针进行筛选。数据将用最好的管家基因进行归一化和分析。然后，将开发一个原型分类模型，并使用从其他正常(n=10)、BD(n=10)和BC(n=10)捐赠者收集的新样本进行验证。数据分析：将进行描述性、图形和非参数统计，以确定生物标志物的表达模式和意义。原型分类模型将通过计算性能评估指标(敏感度、SN、特异度、SP和准确度)进行评估，以区分高性能分类器和无显著分类器的零期望。将在一系列决策规则中报告SN和SP值，以生成接收器操作员特征(ROC)。我们还将评估：a)检测灵敏度，b)分析范围，c)精密度，d)变量之间的相对准确性和折叠变化相关性。预期结果：如果成功，拟议中的血液测试将增强乳房X光检查，更快地产生结果，减少患者在得到结论诊断之前必须等待的时间，并允许远程进行筛查。从长远来看，这样的测试将降低患者的死亡率/发病率和整体医疗成本。公共卫生相关性：虽然乳房X光照相仍然是筛查和检测乳腺疾病的最佳工具，但它可能会漏掉10%-15%的早期癌症，并且无法确定地区分良恶性病变。我们建议开发一种基于血液的测试，这种测试将补充/增强乳房X光检查，产生更快和更客观的结果，减少患者在得到最终诊断之前必须等待的时间，并允许更多的妇女，包括那些生活在农村和医疗服务不足地区的妇女，进行乳腺癌常规检查。这项测试的保健效益将包括降低乳腺癌的发病率/死亡率和总体保健成本/负担。

DESCRIPTION (provided by applicant): Mammography is the benchmark for breast disease detection and diagnosis. However, it can miss 10-15% of early stage breast cancers, and it is unable to distinguish between benign and malignant lesions with certainty. Availability of an alternate method e.g. biomarker based, which allows early detection/precise distinction between benign disease and breast cancer will reduce mortality associated with breast cancer. We previously used DNA microarrays to screen >300 blood RNA specimens, and identified 24 RNA signatures that allowed precise identification of selected patients with breast disease. Based on the prospect of commercializing the identified biomarkers as a blood-based test for breast disease detection, we propose to validate the candidate biomarkers using a platform that is more amenable to translation into the clinic. Objective: To develop a biomarker based blood test for breast disease detection and classification. The specific aims of the project are: 1) design and construct QuantiGene probes for multiplexed blood RNA analysis, 2) test the probes with blood RNA samples, and develop a prototype classification model for identification of individuals with benign breast disease (BD) and breast cancer (BC) and 3) validate the performance of the prototype model in identifying donor categories. Methods: QuantiGene RNA probes targeting 24 biomarkers of interest and 3 housekeeping genes will be designed and constructed. Blood will be collected with PAXGene RNA stabilization tubes, from female donors (>21y) classified as normal (n=30), with BD (n=30) and BC (n=30), and screened with the QuantiGene probes. Data will be normalized with the best housekeeping gene and analyzed. Then, a prototype classification model will be developed and validated using new samples collected from additional normal (n=10), BD (n=10) and BC (n=10) donors. Data analysis: Descriptive, graphical and non-parametric statistics will be performed to determine the pattern and significance of expression of the biomarkers. The prototype classification model will be evaluated by calculating performance evaluation measures (sensitivity, SN; specificity, SP; and accuracy), to distinguish between high-performance classifiers and the null expectation of no significant classifier. SN and SP values will be reported across a range of decision rules to generate the receiver operator characteristics (ROC). We will also assess; a) detection sensitivity, b) assay range, c) precision, d) relative accuracy and fold-change correlations among the variables. Expected outcome: If successful, the proposed blood test will augment mammography, produce faster results, reduce the time a patient has to wait before getting a conclusive diagnosis and allow screening to be performed remotely. In the long run, such a test will reduce patient mortality/morbidity and overall healthcare cost. PUBLIC HEALTH RELEVANCE:Although mammography is still the best tool for screening and detection of breast disease, it can miss 10-15% of early stage cancers and it is unable to differentiate between benign and malignant lesions with certainty. We propose to develop a blood based test that will complement/augment mammography, produce faster and more objective results, reduce the time a patient has to wait before getting a conclusive diagnosis and allow many more women including those in rural and medically underserved areas to routinely test for breast cancer. The health care benefit of this test will include reduced morbidity/mortality and overall healthcare cost/burden due to breast cancer.