Transcriptional effects and non-transcriptional effects of estrogen in the pathogenesis of Inflammatory Breast Cancer

雌激素在炎症性乳腺癌发病机制中的转录作用和非转录作用

基本信息

批准号：
10057443
负责人：
Esther A Peterson
金额：
$ 37.98万
依托单位：
UNIVERSITY OF PUERTO RICO RIO PIEDRAS
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-08-01 至 2023-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10057443
关键词：
3-Dimensional Adopted Affect Animal Model Binding Biological Assay Breast Cancer Cell Breast Cancer cell line Breast cancer metastasis Cancer Prognosis Cell Culture System Cell Line Cell Nucleus Cell Proliferation Cell Survival Cell physiology Cells ChIP-seq Complex Cytoskeleton DNA Binding DNA Binding Domain Data Development Disease ERBB2 gene Embolism Epidermal Growth Factor Receptor Epidermal Growth Factor Receptor Tyrosine Kinase Inhibitor Estradiol Estrogen Receptor alpha Estrogen Receptors Estrogen Therapy Estrogen receptor positive Estrogens GPER gene Gene Expression Genes Genetic Genetic Transcription Goals Growth Factor Receptors Hormonal Human In Vitro Investigation Knowledge Laboratories Ligand Binding Domain Ligands Longitudinal Studies MAP Kinase Gene MAPK3 gene Maintenance Mediating Microscopy Modeling Molecular Neoplasm Metastasis Nuclear Localization Signal Oncogenic Outcome PI3K/AKT Pathogenesis Pathway interactions Patients Pharmaceutical Preparations Phenotype Phosphorylation Phosphotransferases Protein Isoforms Proteins Proto-Oncogene Proteins c-akt RBM5 gene Receptor Activation Receptor Protein-Tyrosine Kinases Research Role Series Signal Pathway Signal Transduction Survival Rate System Techniques Technology Testing Therapeutic Treatment Efficacy breast cancer progression cancer subtypes cell motility cytotoxicity design hormone therapy improved in vitro Model in vivo inflammatory breast cancer inhibitor/antagonist innovation knock-down malignant breast neoplasm migration monocyte new therapeutic target non-genomic novel novel therapeutics outcome forecast overexpression p38 Mitogen Activated Protein Kinase preference receptor response small molecule stem stemness targeted treatment therapeutic target three dimensional cell culture tool transcriptome transcriptome sequencing triple-negative invasive breast carcinoma tumor tumor microenvironment two-dimensional

项目摘要

Project Summary Inflammatory breast cancer (IBC) is one of the most aggressive and lethal form of breast cancer. The molecular mechanisms underlying IBC are poorly understood. Therapeutic strategies have been adopted from non-IBC breast cancers and even though outcomes have been improved for HER2-positive and hormonal receptor IBC, prognosis for triple-negative (TN) subtype IBCs (ER-/PR-/HER2-) is still dire. Currently, estrogen non-genomic signaling has been associated with progression, motility, and invasion of TN breast cancer and most recently of IBC. This proposal seeks to elucidate the functional role of estrogen signaling in IBCs and to compare the effects of estrogen in two contexts: HER2-positive and triple-negative IBC subtypes. Analyses will be performed to define the effects of estrogen signaling, regulated by ERα36 and GPR30, and the cross- activation of ErbB signaling important in the acquisition of various oncogenic phenotypes. The target cells for analysis will be IBC cell lines, SUM149 (TNBC) and SUM190 (HER2+). Around 40% of IBC are TN breast cancers making it hard to treat with endocrine therapy, which is quite effective in ER+/PR+ BC subtypes. IBC shows over-expression of ErbB tyrosine kinase receptors, especially EGFR, in about 50% of the cases, but targeted therapy against this receptor has not been effective for this disease. A series of mechanistic studies will be performed to evaluate the effects of estrogen in the acquisition of oncogenic phenotypes upon knockdown or over-expression of the alternative estrogen receptors, and activation or inhibition of the estrogen signaling. IBC cell lines will be treated with ligands and inhibitors (estradiol,G1,G15, Icaritin) specific to the estrogen receptors to then thoroughly characterized the activation of EGFR downstream kinases and expression of effector proteins and transcriptional changes associated with estrogen signaling. After identifying kinases activated by estrogen signaling, drug response analyses will be done to test novel therapeutic targets with single agents or in combination with EGFR inhibitors. The effects of estrogen non-genomic signaling in pro-oncogenic phenotypes (proliferation, motility, and invasion) will be evaluated using a novel invasion assay and three-dimensional culture system that mimics the formation of IBC tumor emboli. Also, we will determine the DNA binding preferences of ERα36, due to the fact that this isoform of ERα conserved the ligand binding domain, DNA binding domain and nuclear localization signal. In parallel, by RNA-seq analysis we will determine the transcriptome changes associated with estrogen treatment in our two cell line models to identify pathways affected by estrogen relevant in motility, invasion and/or stemness. The poor prognosis for patients with IBC emphasizes the need to further characterize the functional changes associated with its aggressive progression and the interaction of estrogen non-genomic signaling with EGFR pathway. In the long-term, this study will help us design more effective targeted therapies that can be tested in animal models and understand further the mechanisms associated with IBC progression.

项目概要炎性乳腺癌 (IBC) 是最具侵袭性和致命性的乳腺癌之一。人们对 IBC 的分子机制知之甚少。非 IBC 乳腺癌，尽管 HER2 阳性和激素治疗的结果已得到改善受体 IBC，三阴性（TN）亚型 IBC（ER-/PR-/HER2-）的预后目前仍然很糟糕。非基因组信号传导与 TN 乳腺癌的进展、运动和侵袭相关该提案旨在阐明雌激素信号在 IBC 中的功能作用。比较雌激素在两种情况下的影响：HER2 阳性和三阴性 IBC 亚型。进行以确定由 ERα36 和 GPR30 调节的雌激素信号传导的影响，以及交叉 ErbB 信号传导的激活对于获得各种致癌表型非常重要。分析将采用 IBC 细胞系、SUM149 (TNBC) 和 SUM190 (HER2+)。大约 40% 的 IBC 是 TN 乳腺癌。癌症使得内分泌疗法难以治疗，而内分泌疗法对 ER+/PR+ BC 亚型非常有效。 IBC 在大约 50% 的病例中显示 ErbB 酪氨酸激酶受体过度表达，尤其是 EGFR，但针对该受体的靶向治疗尚未对这种疾病有效。将进行评估雌激素在敲低后获得致癌表型的影响或替代雌激素受体的过度表达，以及雌激素信号传导的激活或抑制。 IBC 细胞系将用雌激素特异性配体和抑制剂（雌二醇、G1、G15、淫羊藿素）处理受体，然后彻底表征 EGFR 下游激酶的激活和表达鉴定激酶后与雌激素信号传导相关的效应蛋白和转录变化。由雌激素信号激活，将进行药物反应分析以测试新的治疗靶点与单一药物或与 EGFR 抑制剂联合使用雌激素非基因组信号传导的影响。将使用新的侵袭来评估促癌表型（增殖、运动和侵袭）模拟 IBC 肿瘤栓子形成的测定和三维培养系统此外，我们还将。确定 ERα36 的 DNA 结合偏好，因为该 ERα 亚型保守配体同时，通过 RNA-seq 分析，我们将结合域、DNA 结合域和核定位信号。在我们的两个细胞系模型中确定与雌激素治疗相关的转录组变化，以识别雌激素影响与运动、侵袭和/或干性相关的途径。患者预后不良。 IBC 强调需要进一步描述与其攻击性相关的功能变化从长远来看，雌激素非基因组信号传导与 EGFR 通路的相互作用。这项研究将帮助我们设计更有效的靶向疗法，可以在动物模型中进行测试进一步了解与 IBC 进展相关的机制。