Mechanistic Links Between Changing Estrogen Profiles, Inflammation and the Increased Risk and Metastasis of Breast Cancer in Obese Women

肥胖女性雌激素水平变化、炎症与乳腺癌风险增加和转移之间的机制联系

• 批准号: 10585320
• 负责人: JOYCE MARIE SLINGERLAND
• 金额: $ 41.52万
• 依托单位: GEORGETOWN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-03-07 至 2027-11-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10585320
• 关键词: 3-Dimensional; Adipocytes; American; Binding; Breast; Breast Cancer Cell; Breast Cancer Risk Factor; Breast Cancer cell line; Breast Cancer therapy; Breast cancer metastasis; CCL2 gene; Cells; ChIP-seq; Chromatin; Chronic; Cytokine Gene; Data; Development; Drug resistance; ESR1 gene; Estradiol; Estrogen receptor positive; Estrogens; Estrone; Euchromatin; Expression Profiling; Fatty acid glycerol esters; Gene Activation; Gene Expression; Gene Expression Profile; Gene Expression Regulation; Genes; Genetic Transcription; Grant; Growth; Heterochromatin; Hormones; Human; IL6 gene; In Vitro; Inflammation; Inflammatory; Ligands; Link; MCF7 cell; Malignant Neoplasms; Mammary Neoplasms; Mediating; Molecular Conformation; Mutation; Neoplasm Metastasis; Nuclear; Obesity; Obesity Epidemic; Oncogenes; Oncogenic; Organoids; Ovarian; Postmenopause; Premenopause; Prevalence; Prevention strategy; Prognosis; Regulation; Repression; Repressor Proteins; Response Elements; Risk; Sampling; Site; Testing; Thinness; Tissues; Trans-Activators; Tumor Promotion; Woman; Work; Xenograft procedure; cancer cell; cancer prevention; cancer stem cell; cell type; cytokine; epithelial to mesenchymal transition; falls; gene induction; gene repression; genetic corepressor; hormone therapy; in vivo; malignant breast neoplasm; mammary; matrigel; mortality; mutant; new therapeutic target; novel; p65; programs; receptor binding; recruit; response; stem cell expansion; transcription factor; transcriptome; transcriptome sequencing; tumor; tumor growth; tumor initiation; tumor progression; tumorigenesis

Obesity prevalence is >39% in the USA. Obesity increases estrone synthesis in fat; and both obesity and estrone correlate with increased postmenopausal ER+ breast cancer risk and mortality. We aim to elucidate how estrone links obesity, inflammation and breast cancer. Obese fat is chronically inflamed via NFB activation. We found breast fat inflammation increases with obesity, after menopause and surrounding cancers. Breast cancer:adipocyte contact upregulates proinflammatory cytokines that stimulate NFB- and estrone:ER-dependent cancer stem cell (CSC) expansion. We showed the dominant premenopausal estradiol (E2) and postmenopausal estrone (E1) regulate different genes. While E2-bound ER inhibits NFB, we showed E1-bound ER is an NFB co-activator and induces gene profiles of inflammation, EMT, CSC expansion and metastasis, while E2 did not. Finally, E1: ER caused more cytokine gene induction, stem cell expansion, and ER+ tumor growth and metastasis than E2 in vivo. While our in vivo data suggest E1 driven ER-NFB co-targets promote tumor progression, little is known about how E1-bound ER and NFB interact at chromatin, and how their co-regulators differ from those of E2-ER. Up to 30% of metastatic ER+ BC develop activating ESR1 mutations. We found while 3D growth of MCF7 controls is greater with E1 than E2; both stimulate 3D growth of ER mutant MCF7 lines equally. Further, both ER mutants direct greater NFB activation upon either E1 or E2 treatment, suggesting that the altered mutant ER conformation might direct greater ER-p65B activation. We hypothesize that E1-ER target gene selection, co-regulators, and expression differ from those of E2:ER and that therapy-induced ER mutants will permit pro-inflammatory, oncogenic, and prometastatic ER-NFB target genes, normally activated only by E1-bound ER-WT, to be indiscriminately activated by either E1 or E2 in cancers. Aim 1 will test how E1 and E2 driven gene expression differs, comparing ChIPseq of ER, p65B and FOXA1 and correlating these with gene expression profiles in ER+ cancer lines and organoids stimulated by E1 vs E2, +/- NFB. Aim 2 To identify co-regulators unique to E1 and E2-liganded ER that mediate gene induction or repression, we carry out i) ChIP-Mass Spec in cells treated with E1 or E2, +/- NFB activation; and ii) Gradient-Seq to separate euchromatin from heterochromatin followed by ChIPseq and ChIP-Mass Spec to identify the E1- vs E2-liganded ER interactome in transactivator versus repressor complexes. Aim 3 will test if both E1 and E2 i) cause greater ER:p65 binding and ii) preferential activation of oncogenic ER mutant: B co-target genes normally activated by E1-liganded ER-WT +/-NFB in vitro, and ii) activate a more E1-ER-like oncogenic genes profile in ERY537S BC xenografts and PDX than in ER-WT BC tumors in vivo. This will inform how ER target gene changes during the shift from high E2 to high E1 after menopause might promote breast cancer development and may identify new therapeutic targets, ultimately yielding new ER+ breast cancer therapies and prevention strategies.

在美国，肥胖率>39% 肥胖会增加脂肪中的雌酮合成； 雌酮与绝经后 ER+ 乳腺癌风险和死亡率增加相关。 雌酮如何与肥胖、炎症和乳腺癌联系起来 肥胖脂肪通过 NFB 慢性发炎。 我们发现乳房脂肪炎症随着肥胖、绝经后及其周围环境的增加而增加。 乳腺癌：脂肪细胞接触上调促炎性细胞因子，刺激 NFκB- 和 雌酮：ERα依赖性癌症干细胞（CSC）扩增我们显示了绝经前的显性扩增。 雌二醇 (E2) 和绝经后雌酮 (E1) 调节不同的基因，而 E2 结合的 ERα 抑制 NFβB， 我们发现 E1 结合的 ERα 是一种 NFβ 共激活剂，可诱导炎症、EMT、CSC 的基因谱 扩增和转移，而E2没有。最后，E1：ERα引起更多的细胞因子基因诱导，干细胞。 在体内，ER+ 的肿瘤生长和转移优于 E2，而我们的体内数据表明 E1 驱动。 ER-NFB 共同靶点促进肿瘤进展，但人们对 E1 结合的 ER 和 NFB 如何相互作用知之甚少。 染色质，以及它们的共同调节因子与 E2-ER 的不同之处 高达 30% 的转移性 ER+ BC 发生。 我们发现，E1 的 MCF7 对照的 3D 生长比 E2 更大； 同样地刺激 ER 突变体 MCF7 的 3D 生长 此外，两种 ER 突变体都指导更大的 NF+B。 E1 或 E2 处理后激活，表明突变 ER 构象的改变可能会指导 我们追踪了 E1-ERα 靶基因选择、协同调节因子和更大的 ER-p65β 激活。 表达与 E2:ER​​ 不同，并且治疗诱导的 ER 突变体将允许促炎、 致癌和促转移 ERα-NFβB 靶基因，通常仅由 E1 结合的 ER-WT 激活， 目标 1 将测试 E1 和 E2 如何驱动基因表达。 不同的是，比较了 ER、p65+B 和 FOXA1 的 ChIPseq，并将它们与基因表达谱相关联 E1 与 E2 刺激的 ER+ 癌系和类器官，+/- NF+B 目标 2 识别独特的共同调节因子。 E1 和 E2 配体 ERα 介导基因诱导或抑制，我们在细胞中进行 i) ChIP-Mass Spec 用 E1 或 E2 处理，+/- NF+B 激活；和 ii) 梯度测序以分离常染色质和异染色质 随后使用 ChIPseq 和 ChIP-Mass Spec 来鉴定反式激活因子中 E1 与 E2 配体的 ERα 相互作用组 目标 3 将测试 E1 和 E2 是否 i) 引起更大的 ER:p65 结合和 ii) 优先激活致癌 ER 突变体：通常由 E1 配体 ER-WT 激活的B 共靶基因 +/-NFB 体外，以及 ii) 在 ERY537S BC 异种移植物和 PDX 中激活更像 E1-ER 的致癌基因谱 与体内 ER-WT BC 肿瘤相比，这将了解 ER 靶基因在从高 E2 转变为高 E2 期间如何变化。 绝经后高 E1 可能会促进乳腺癌的发展，并可能确定新的治疗靶点， 最终产生新的 ER+ 乳腺癌治疗和预防策略。