Proteomic Identification of NAF Biomarkers

描述（由申请人提供）：乳腺癌每年造成约43,000人死亡，如果及早发现这种疾病，可以大大减少这一数字。目前，乳腺癌的检测主要依靠身体检查和常规乳房x光检查。虽然这些程序改善了乳腺癌的早期发现，从而降低了死亡率，但它们仍然导致假阳性和假阴性诊断率相对较高。此外，乳腺癌的预后是基于组织学检查，不足以评估微转移。由于无法准确预测复发风险，50%或更多的乳腺癌患者不必要地接受辅助治疗。如果这些现有的基于观察的方法与分子评估相辅相成，可以获得更有用和准确的乳腺癌评估。据报道，与健康个体相比，乳腺癌患者的许多血清蛋白发生了改变。然而，目前还没有任何已知的循环蛋白可以用来确定乳腺癌的阶段或作为乳腺癌的一般标志，特别是在这种疾病的早期阶段。乳腺导管细胞是70%至80%乳腺癌病例的细胞起源。乳头吸出液（NAF）来自非哺乳期妇女，含有由这些导管细胞直接分泌的蛋白质。因此，我们假设NAF是乳腺癌蛋白质生物标志物的浓缩和选择性来源。蛋白质组学方法提供了一种公正的方法来评估NAF作为生物标志物的来源，并且对小NAF体积（10至50 (l)）足够敏感。因此，本提案的目标是表征NAF的蛋白质组，并进行初步分析以确定潜在的蛋白质生物标志物。为实现这些目标，我们提出以下具体目标：目标1。用质谱法表征合并NAFs的蛋白质组。目标# 2。利用同位素标记和质谱技术鉴定早期乳腺癌女性NAF样本中潜在的蛋白质生物标志物在完成这些目标后，我们将提供NAF蛋白质组的第一个详细特征，并确定了NAF蛋白质的一个子集，该子集是乳腺疾病的潜在标记物。

DESCRIPTION (provided by applicant): Breast cancer is responsible for ~43,000 deaths per year, a number which could be reduced dramatically by early detection of the disease. Currently, detection of breast cancer relies primarily on physical examination and conventional mammography. While these procedures have improved early detection of breast cancer and thereby decreased mortality, they still result in relatively high rates of false-positive and false-negative diagnoses. In addition, breast cancer prognosis is based on histological examination and is inadequate for assessing micrometastases. Due to this inability to accurately predict the risk of recurrence, 50% or more of breast cancer patients are unnecessarily treated with adjuvant therapy. A more useful and accurate evaluation of breast cancer could be obtained if these current, observation-based methods were supplemented with a molecular assessment. A number of serum proteins are reported to be altered in individuals with breast cancer when compared with healthy individuals. There is not, however, any known circulating protein that is suitable to define the stage of breast cancer or as a general marker for breast cancer, especially in the early stages of this disease. Mammary ductal cells are the cellular origin for 70% to 80% of breast cancer cases. Nipple aspirate fluid (NAF), which is obtained from non-lactating women, contains proteins secreted directly by these ductal cells. As such, we hypothesize that NAF is a concentrated and selective source of protein biomarkers for breast cancer. Proteomic approaches offer an unbiased way to evaluate NAF as a source of biomarkers and is sufficiently sensitive for small NAF volumes (10 to 50 (l). Therefore, the goals of this proposal are to characterize the proteome of NAF and to undertake preliminary analyses to identify potential protein biomarkers. To accomplish these goals, we propose to undertaken the following specific aims: Aim #1. Characterize the Proteome of Pooled NAFs Using Mass Spectrometry. Aim #2. Identify Potential Protein Biomarkers in Individual NAF Samples from Women with Early-Stage Breast Cancer using Isotopic Labeling and Mass Spectrometry. Upon completion of these aims, we will have provided the first detailed characterization of the NAF proteome and identified a subset of NAF proteins that are potential markers of breast disease.