Discovery and validation of early molecular breast cancer risk markers in benign breast disease

项目总结 在美国，每年有160万女性因良性乳腺疾病(BBD)而接受乳腺活检。90% 其中没有显示出细胞异型性的证据，只是略有增加患乳腺癌的风险，但这一点 这一群体代表了患有BBD的女性中患癌症的绝大多数，因为患有BBD的女性 可识别的风险因素，如非典型性或强阳性家族史，是一小部分。BBD中的病变 后来发展为癌症的女性很可能表现出比明显的癌症更早的分子变化 好几年了。检测到这种变化的化验将对我们准确识别的能力提供关键的改进 预防干预的高危妇女(特别是在区分雌激素受体(ER)风险的情况下 阳性乳腺癌与阴性乳腺癌)，并允许更好地针对以下女性的高级监测策略 将受益最大。启动子异常甲基化在癌症进展过程中很早就被证实。 我们已经成功地将询问甲基组的全基因组阵列工具调整为福尔马林固定的 组织样本，创造了研究具有已知长期结果的档案样本的机会。 在过去的15年里，我们已经建立了一个完整的临床-病理关系数据库，其中包括所有电子数据 自1985年以来，该机构有超过75,000名乳房患者。我们发现患有非典型BBD的女性 随后患上乳腺癌(病例)，并将它们与患有BBD的女性配对，后者仍患有癌症- 免费(对照)，有文件记录的至少10年的随访。在一项初步研究中，我们证明了 BBD档案组织中的分子特征与随后的浸润性乳腺癌中的分子特征相匹配 (IBC)；此外，这些特征与对照组BBD患者不同，没有证据表明 随后的IBC，ER+与ER-病例不同。 我们现在建议将我们的科学和临床资源结合起来，使风险的发现和验证成为可能 源自BBD的标记。我们已经确认了500多例早于IBC和配对IBC的BBD病例 已知ER状态的样本和可用的档案组织块。我们将进行一项病例对照研究 185例非典型非家族性BBD及其继发的IBC的甲基组研究 对照样本，与随访、年龄和是否存在增生相匹配。我们还将捕获已知风险 评估末端导管小叶单位退缩的IBC、乳腺X光密度和体重指数的因素 (TDLU)，并用免疫组织化学半定量方法鉴定关键激素受体的状态。 我们的假设是，BBD在IBC发展之前就存在分子变化，并且是明显的 在随后发展为ER+与ER-IBC的女性中。我们的首要目标是确定 乳腺组织中的分子变化将使所有接受乳房手术的女性能够准确地进行风险分层 活组织检查显示没有异型性。在未来，这样的生物标记物可能适用于极少量的乳房样本 组织，如通过随机细针抽吸或在乳头液中获得的组织。

PROJECT SUMMARY 1.6 million women undergo breast biopsy for benign breast disease (BBD) annually in the United States. 90% of these show no evidence of cellular atypia, and are at only modestly increased risk for breast cancer, but this group represents the great majority who develop cancer among women with BBD, since women with identifiable risk factors, such as atypia or strongly positive family history are a small minority. BBD lesions in women who later progress to cancer very likely display molecular changes which predate overt cancer by years. An assay detecting such changes will provide a critical improvement in our ability accurately identify high risk women for preventive interventions (particularly if it distinguishes risk of estrogen receptor (ER) positive vs. negative breast cancer), and allow better targeting advanced surveillance strategies to women who will benefit the most. Aberrant promoter methylation has been documented very early in cancer progression. We have successfully adapted genome-wide array-based tools interrogating the methylome to formalin-fixed tissue samples, creating an opportunity to study archival samples with known long-term outcomes. For the past 15 years, we have built an integrated clinical-pathological relational database of all electronic data of over 75,000 breast patients at this institution since 1985. We identified women with non-atypical BBD who subsequently developed breast cancer (cases), and matched them to women with BBD who remained cancer- free (controls) with documented follow-up of a minimum of 10 years. In a pilot study, we demonstrated that molecular signatures in archival BBD tissue matched signatures present in subsequent invasive breast cancer (IBC); furthermore, these signatures were distinct from those in control BBD patients with no evidence of subsequent IBC, and differed in ER+ vs ER- cases. We now propose to combine our scientific and clinical resources to enable the discovery and validation of risk markers derived from BBD. We have identified over 500 cases with BBD predating IBC and paired IBC samples with known ER status with available archival tissue blocks. We will perform a case-control study of the methylomes of archival non-atypical non-familial BBD from 185 cases as well as their subsequent IBC, and 75 control samples, matched by follow-up, age and presence of hyperplasia. We will also capture known risk factors for IBC, mammographic density and body mass index, assess involution of terminal duct lobular units (TDLU), and characterize the status of key hormone receptors by semiquantitative immunohistochemistry. Our hypothesis is that molecular changes are present in BBD prior to the development of IBC, and are distinct in women who subsequently develop ER + versus ER- IBC. Our overarching goal is the identification of molecular changes in breast tissue that will allow the accurate risk stratification of all women undergoing breast biopsy showing no atypia. In the future, such biomarkers may be applicable to minimal samples of breast tissue such as those obtained with random fine needle aspiration or in nipple fluid.