Project 2: Inflammation-Based Mechanisms of Hormone Therapy Resistance in Breast Cancer

项目2摘要 乳腺癌是女性癌症死亡的第二大原因，占乳腺肿瘤的近75%。 在诊断时表达雌激素受体-α(ER)。据估计，大约30%-40%的ER+乳房 肿瘤对激素治疗产生抵抗力，要么是从头产生的，要么是获得性的。最近的证据 提示炎症在激素获得性抵抗和发病机制中起关键作用 治疗，并被认为是乳腺癌的风险因素。为此，我们和其他人已经发现了一个 乳腺癌炎症信号与内分泌耐药的重要联系机制 ER和NFB之间的相互作用，通过细胞因子诱导的ER的磷酸化和直接调节 先锋因子FOXA1。这一点很重要，因为超过80%的淋巴转移和65%- 70%来自ER+原发肿瘤的远处转移在复发时保留ER的表达。另外， 炎性介质，如细胞因子，如肿瘤坏死因子或主要转录因子 核因子B及其上游调节因子IKK在乳腺肿瘤中高度表达，并随着肿瘤分级的增加而增加。 我们将使用ER+乳腺癌患者来源的异种移植物(PDX)和分离的FACS(EpCAM+/CD49f)ER+ 从多个患者分离腔细胞，以了解炎症信号如何影响ER功能。我们的 假说是，由核因子B及其上游调节因子IKK驱动的炎症信号导致内质网改变 功能和靶基因表达导致更具侵袭性的肿瘤和对 内分泌治疗。首先，我们将定义原发肿瘤的转录和表观遗传反应。 利用新的下一代测序技术检测雌激素和细胞因子诱导的炎症信号 技术(如GRO-SEQ，ATAC-SEQ和CHIP-SEQ)，然后我们将测试原发人类肿瘤 标本，表现出更多的增殖，侵袭，转移和对激素治疗的抵抗 暴露于雌激素和炎性细胞因子。我们还将测试新的IKK/NFB抑制剂是否会拯救 肿瘤标本对激素治疗的敏感性。对我们生成的数据进行生物信息学分析 许多患者将允许我们识别潜在的预后的生物标记物和/或基因签名。 内分泌治疗结果较差的患者和/或可能受益于 核因子B信号的抑制剂。下面列出了我们三个具体目标的标题： 目的1：研究原代人雌激素受体和核因子B的转录变化及染色质结合谱 组织对雌二醇和促炎细胞因子的反应。 目的2：确定炎症调节ER功能在乳腺肿瘤中的生物学后果 目标3：定义预后标志，确定生物标记物，并测量基于炎症的患病率 跨患者样本的ER功能的调制。

Project 2 Abstract Breast cancer is the second leading cause of cancer mortality in women and nearly 75% of breast tumors express estrogen receptor-α (ER) at the time of diagnosis. It is estimated that about 30-40% of ER+ breast tumors become resistant to hormone therapy, either through de novo or acquired resistance. Recent evidence suggests that inflammation plays a key role in promoting pathogenesis and acquired resistance to hormone therapies, and is considered a risk factor for breast cancer. To this end, we and others have uncovered an important mechanism linking inflammatory signaling to endocrine resistance in breast cancer through interactions between the ER and NFB, via cytokine-induced phosphorylation of ER and direct modulation of the ER pioneer factor FOXA1. This is important because greater than 80% of the lymph node metastases and 65– 70% of distant metastases arising from ER+ primary tumors retain ER expression at the time of relapse. Also, inflammatory mediators, such as cytokines like the tumor necrosis factor alpha (TNF) or the master transcription factor NFB and its upstream regulator IKK, are highly present in breast tumors and increase with tumor grade. We will use ER+ breast cancer patient-derived xenografts (PDXs), and FACS isolated (EpCAM+/CD49f) ER+ luminal cells isolated from multiple patients, to understand how inflammatory signaling affects ER function. Our hypothesis is that inflammatory signaling, driven by NFB and its upstream regulator IKK, leads to altered ER function and target gene expression resulting in more aggressive tumors and an increased resistance to endocrine therapy. First we will define the transcriptional and epigenetic response of the primary tumor specimens to estrogen and cytokine induced inflammatory signaling using novel next-generation sequencing technologies (such as GRO-seq, ATAC-seq and ChIP-seq) Then we will test if the primary human tumor specimens, exhibit increased proliferation, invasion, metastasis and resistance to hormone therapy when exposed to estrogen and inflammatory cytokines. We will also test if novel inhibitors of IKK/NFB will rescue sensitivity to hormone therapy in the tumor specimens. Bioinformatic analysis of the data we generate across many patients will allow us to identify biomarkers and/or gene signatures that are potentially prognostic of patients that have worse outcomes on endocrine therapies and/or predictive of patients who may benefit from inhibitors of NFB signaling. Listed below are the headings for our 3 specific aims: Aim 1:Define the transcriptional changes and chromatin binding profiles of ER and NFB in primary human tissues in response to E2 and proinflammatory cytokines. Aim 2:Determine the biological consequences of inflammation-based modulation of ER function in breast tumors Aim 3:Define prognostic signatures, identify biomarkers and measure prevalence of inflammation-based modulation of ER function across patient samples.