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，肥胖症患病率为39%。肥胖增加脂肪中雌酮的合成；肥胖和 雌酮与绝经后ER+乳腺癌风险和死亡率的增加有关。我们的目标是澄清 雌酮如何将肥胖、炎症和乳腺癌联系起来。肥胖的脂肪通过核因子B慢性发炎 激活。我们发现，随着肥胖、绝经后和周围环境的增加，乳房脂肪炎症会增加 癌症。乳腺癌：脂肪细胞接触上调促炎症细胞因子，刺激NF-B-和 雌酮：ER-依赖的肿瘤干细胞的扩增。我们展示了绝经前的主要症状 雌二醇(E2)和绝经后雌酮(E1)调节不同的基因。而E2结合的ER抑制核因子B， 我们发现，E1结合的ER是一种核因子B共激活因子，可诱导炎症、子宫内膜癌、中枢神经干细胞的基因图谱。 扩张和转移，而E2无此作用。最后，E1：ER引起了更多的细胞因子基因诱导，干细胞 在体内，ER+肿瘤的生长和转移优于E2。虽然我们的活体数据表明 ER-NFB共靶点促进肿瘤进展，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 激活ESR1突变。我们发现，虽然使用E1的MCF7对照组的3D增长比使用E2的更快；但两者都 同样刺激ER突变型MCF7细胞系的三维生长。此外，两个ER突变体都指示更大的NFB 在E1或E2处理时激活，表明改变的突变ER构象可能直接 更强的ER-p65B活性。我们假设，E1-ER的靶基因选择、协同调节和 与E2：ER的表达不同，治疗诱导的ER突变将允许促炎， 致癌和转移性ER-NFB靶基因，通常仅由E1结合的ER-WT激活，将被 在癌症中被E1或E2不分青红皂白地激活。Aim 1将测试E1和E2如何驱动基因表达 不同，比较ER，p65B和FOXA1的ChIPseq，并将它们与 E1vsE2，+/-NFB刺激的ER+癌细胞株和有机体。目的2鉴定独有的共同调节因子 E1和E2连接的ER介导基因诱导或抑制，我们在细胞中进行了I)芯片-质谱仪 用E1或E2处理，+/-NFB激活；以及ii)梯度序列分离常染色质和异染色质 CHIPSEQ和CHIP-MS鉴定反式激活子中的E1-VS E2连接的ER-相互作用组 而不是阻遏复合体。目标3将测试E1和E2是否都会导致更大的ER：P65结合和II) 癌基因ER突变体的优先激活：B共靶基因通常由E1连接的ER-WT激活 +/-NFB，以及II)在ERY537S BC异种移植瘤和PDX中激活更多的E1-ER样癌基因 在体内ER-WT BC肿瘤中的表达高于ER-WT BC肿瘤。这将告知ER靶基因在从高E2到 绝经后高E_1可能促进乳腺癌的发展，并可能寻找新的治疗靶点。 最终产生新的ER+乳腺癌治疗和预防策略。