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> 39％。肥胖会增加脂肪中的雌酮合成。以及肥胖和 雌酮与绝经后ER+乳腺癌的风险和死亡率相关。我们旨在阐明 雌酮如何联系肥胖，感染和乳腺癌。肥胖脂肪长期通过NFB发炎 激活。我们发现乳腺脂肪感染随肥胖，更年期和周围的肥胖增加而增加 癌症。乳腺癌：脂肪细胞接触上调刺激NFB-和的促炎细胞因子 雌酮：ER依赖性癌症干细胞（CSC）膨胀。我们展示了主要的绝经前 雌二醇（E2）和绝经后雌酮（E1）调节不同的基因。而e2结合的er抑制了nFB，但 我们显示E1结合的ER是NFB共激活因子，影响炎症，EMT，CSC的基因谱。 扩展和转移，而E2没有。最后，e1：er引起了更多的细胞因子基因诱导，干细胞 与体内E2相比，扩张和ER+肿瘤生长和转移。而我们的体内数据建议E1驱动器 ER-NFB共同目标促进肿瘤的进展，对E1结合的ER和NFB的相互作用知之甚少 染色质，以及它们的共同调节剂与E2-ER的共同调节剂的不同。最多30％的转移性ER+ BC开发 激活ESR1突变。我们发现，虽然E1的3D增长MCF7对照组大于E2。两个都 同样刺激ER突变体MCF7线的3D生长。此外，两个ER突变体导向更大的NFB 在E1或E2处理后激活 更大的ER-P65B激活。我们假设E1-er靶基因选择，共同调节剂和 表达与E2不同的表达不同，而治疗诱导的ER突变体将允许促炎性， 通常仅通过E1结合的ER-WT激活的致癌性和前态ER-NFB靶基因 在癌症中被E1或E2激活。 AIM 1将测试E1和E2驱动的基因表达方式 差异，比较ER，p65b和foxA1的chipseq，并将其与基因表达谱相关联 E1 vs e2刺激的ER +癌品系和类器官。目标2以识别独特的共同调节器 介导基因诱导或表达的e1和e2-rigander，我们在细胞中执行i）chip-mas spec 用E1或E2处理+/-NFB激活； ii）梯度seq以将白染色质与异染色质分开 然后是chipseq和Chip-Mass规格，以识别transactivator中的E1-E2和e2配合的ER相互作用组 与复制复合物。 AIM 3将测试E1和E2 I）是否引起更大的ER：P65结合和II） 致癌ER突变体的优先激活：B共同靶向基因通常被E1配合的ER-WT激活 +/-nFB在体外，ii）激活E1-ER样的致癌基因谱图BC Xenographics和PDX 比在体内的Er-WT BC肿瘤中。这将告知ER靶基因在从高E2转移到的转移期间如何变化 更年期后的高E1可能会促进乳腺癌的发展，并可能确定新的治疗靶标， 最终产生了新的ER+乳腺癌疗法和预防策略。