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

项目摘要 1.6在美国，每年有100万妇女接受乳腺活检以诊断良性乳腺疾病（BBD）。百分之九十 这些人没有表现出细胞增殖的证据，患乳腺癌的风险也只是适度增加，但这 组代表了绝大多数患有BBD的女性中患癌症的人，因为患有BBD的女性 可识别的风险因素，如梅毒或强阳性家族史是少数。BBD病变， 后来发展为癌症的妇女很可能表现出早于明显癌症的分子变化， 年检测这种变化的检测将为我们准确识别 高危妇女进行预防性干预（特别是如果它区分雌激素受体（ER）的风险） 阳性与阴性乳腺癌），并使先进的监测战略更好地针对妇女， 将受益最大。异常启动子甲基化在癌症进展的非常早期就已被证实。 我们已经成功地将基于全基因组阵列的工具用于询问福尔马林固定的甲基化组， 组织样本，创造了一个机会，研究档案样本与已知的长期结果。 在过去的15年里，我们建立了一个所有电子数据的综合临床-病理关系数据库 从1985年开始，这里有超过75,000名乳腺癌患者我们发现患有非典型BBD的女性， 随后发展为乳腺癌（例），并将其与患有BBD的女性进行匹配，这些女性仍然患有癌症- 免费（对照），记录随访至少10年。在一项试点研究中，我们证明， 档案BBD组织中的分子特征与后续侵袭性乳腺癌中存在的特征相匹配 (IBC)此外，这些签名与对照BBD患者的签名不同，没有证据表明 随后的IBC，并且在ER+与ER-病例中存在差异。 我们现在建议联合收割机结合我们的科学和临床资源，使风险的发现和验证 来自BBD的标记。我们已经确定了超过500例早于IBC和配对IBC的BBD病例 已知ER状态的样本和可用的存档组织块。我们将进行一项病例对照研究， 185例非典型非家族性BBD及其随后的IBC的档案甲基化组，和75例非典型非家族性BBD及其随后的IBC的档案甲基化组。 对照样本，通过随访、年龄和增生的存在匹配。我们还将捕捉已知风险 IBC的因素，乳腺摄影密度和体重指数，评估终末导管小叶单位的退化 （TDLU），并通过半定量免疫组织化学表征关键激素受体的状态。 我们的假设是，分子变化存在于BBD之前的IBC的发展， 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，这将允许对所有接受乳腺癌手术的女性进行准确的风险分层。 活组织检查显示没有子宫肌瘤。在未来，这种生物标志物可能适用于乳腺癌的最小样本。 组织，例如用随机细针抽吸或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.