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

项目2 乳腺癌是女性癌症死亡率的第二大原因，占乳腺肿瘤的近75% 在诊断时表达雌激素受体-α（ER）。据估计，约30-40%的ER+乳腺癌患者 肿瘤对激素疗法产生抗性，或者是通过从头产生抗性，或者是通过获得性抗性。最近的证据 表明炎症在促进发病和获得性激素抵抗中起关键作用 乳腺癌的早期症状有哪些？为此，我们和其他人发现了一个 乳腺癌中炎症信号传导与内分泌抵抗的重要机制 ER和NF κ B B之间的相互作用，通过精氨酸诱导的ER磷酸化和直接调节 ER先驱因子FOXA 1。这一点很重要，因为超过80%的淋巴结转移和65%的淋巴结转移都是由淋巴结转移引起的。 70%的ER+原发性肿瘤的远处转移在复发时保留ER表达。还有， 炎症介质，如细胞因子如肿瘤坏死因子α（TNF）或主转录因子， 因子NF κ B B及其上游调节因子IKK在乳腺肿瘤中高度存在，并随肿瘤分级而增加。 我们将使用ER+乳腺癌患者来源的异种移植物（PDX）和FACS分离的（EpCAM+/CD 49 f）ER+细胞（CD 49 f）。 从多名患者中分离的腔细胞，以了解炎症信号如何影响ER功能。我们 一种假说认为，由NF κ B B及其上游调节因子IKK驱动的炎症信号传导导致ER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，我们将定义原发性肿瘤的转录和表观遗传反应 使用新的下一代测序对雌激素和细胞因子诱导的炎症信号传导的标本进行检测 技术（如GRO-seq，ATAC-seq和ChIP-seq），然后我们将测试原发性人类肿瘤是否 标本，表现出增加的增殖，侵袭，转移和抵抗激素治疗时， 暴露在雌激素和炎症细胞因子中我们还将测试新的IKK/NF κ B B抑制剂是否能拯救 对激素治疗的敏感性。生物信息学分析我们产生的数据， 许多患者将使我们能够识别生物标志物和/或基因特征，这些生物标志物和/或基因特征可能预示着 内分泌治疗结局较差的患者和/或预测可能受益于 NF κ B B信号传导抑制剂。以下是我们三个具体目标的标题： 目的1：明确ER和NF-κ B在原代人乳腺癌细胞中的转录变化和染色质结合特征 组织对E2和促炎细胞因子的反应。 目的2：确定乳腺肿瘤中基于炎症的ER功能调节的生物学后果 目的3：定义预后特征，识别生物标志物并测量基于炎症的患病率 调节患者样品中的ER功能。

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.