Roles of E2F3 and SGO1 in the Epithelial-to-Mesenchymal Transition and Metastasis of Breast Cancer

E2F3和SGO1在乳腺癌上皮间质转化和转移中的作用

• 批准号: 9911351
• 负责人: Shirley Jusino-Cotto
• 金额: $ 3.19万
• 依托单位: PONCE SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-30 至 2021-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9911351
• 关键词: 3-Dimensional; Address; African American; Apoptosis; Automobile Driving; Behavior; Binding; Biological Assay; Biological Response Modifier Therapy; Breast Cancer Cell; Breast Cancer Patient; Breast Cancer therapy; Breast Epithelial Cells; Breast cancer metastasis; CRISPR/Cas technology; Cell Cycle; Cell Differentiation process; Cell Line; Cell physiology; Cells; Centrioles; Centrosome; Chromosomal Instability; Complementary DNA; Cytokinesis; DNA biosynthesis; Data; Development; Down-Regulation; E2F1 gene; Event; Family; G1 Phase; Gene Expression; Genes; Genetic Transcription; Genome Stability; Goals; Hispanics; Immunofluorescence Immunologic; Immunohistochemistry; Individual; Injections; Investigation; Knock-out; Lead; MDA MB 231; Maintenance; Malignant Neoplasms; Measures; Mesenchymal; Metastatic breast cancer; Mitosis; Mitotic; Modeling; Mus; Neoplasm Metastasis; Outcome; Pathway interactions; Patients; Primary Neoplasm; Process; Proteins; Publishing; RNA; RPS27 gene; Reporting; Research; Retinoblastoma; Role; Signal Transduction; Small Interfering RNA; Testing; Tissues; Transcription Coactivator; Transcription Repressor; Tumor Burden; Up-Regulation; Weight; Western Blotting; Woman; anticancer research; cancer cell; cancer therapy; design; druggable target; epithelial to mesenchymal transition; experimental study; genome editing; hormone receptor-negative; improved; in vivo; insight; knock-down; malignant breast neoplasm; migration; mouse model; novel; outcome forecast; overexpression; prevent; promoter; retinoblastoma tumor suppressor; small molecule; three dimensional cell culture; tissue culture; transcription factor; triple-negative invasive breast carcinoma; tumor; tumor growth; tumor initiation; tumorigenesis

SUMMARY/ ABSTRACT Despite the great advancements in breast cancer research and treatment, approximately 42,000 US women are expected to die from breast cancer in 2019. Especially deadly are basal, triple- negative (TN) breast cancers. Basal TNBC are more prevalent in African American and Hispanic women and there are no biological therapies available to treat them. The E2 Promoter Binding Factors (E2Fs) are a group of eight transcription factors under retinoblastoma (Rb) control that regulate several cellular processes including DNA replication, genomic stability and apoptosis. Broadly, E2Fs can be divided into transcriptional activators (those that promote gene transcription) and repressors (those that halt gene transcription). Preliminary data I present here shows 67% basal/TN breast cancers overexpress the E2F activators. It is known that the Rb pathway is involved in the epithelial-to-mesenchymal transition (EMT), an early process that lead to metastasis; yet the involvement of the E2F activators driving EMT in breast cancer is still under investigation. Our published data demonstrated that overexpression of E2F activators (E2F1-3) leads to increased expression of mitotic regulators (MPS1, NEK2, SGO1, among others). Furthermore, we recently published that that MPS1 promotes EMT and invasion and NEK2 (an E2F target) generates pre-invasive protrusions in 3D culture models. Hence, we question to what extent deregulation of E2Fs activators promotes EMT, invasion, and metastasis through SGO1. Specifically, we will focus our research in E2F3 because it is overexpressed in 42% basal breast cancers and it we already showed that E2Fs regulate the stability of SGO1 protein levels. Our central hypothesis is that the overexpression of E2F3 contributes to tumorigenesis by promoting EMT through the induction of the transcription of SGO1, thus leading to increased invasion and metastasis. To test our central hypothesis, we will develop the following two specific aims: (1) Determine the effects of E2F3 and SGO1 in EMT, invasion, and migration in metastatic, triple- negative breast cancer cells. (2) Determine the effects of E2F3 knockout in MDA-MB-231 cell lines using CRISPR/Cas9 genome editing in NOD scid gamma mouse models. We expect to identify a mechanism of how E2F3 overexpression promotes EMT through SGO1 in breast cancer, thus contributing to migration, invasion, and metastasis. If the expected outcomes are met, small molecules may be designed to improve current therapies to target downstream effectors of E2F3, since E2Fs cannot be targeted.

总结/摘要 尽管乳腺癌研究和治疗取得了巨大进展，但仍有大约42，000人 预计美国女性将在2019年死于乳腺癌。尤其致命的是基础，三重- 阴性（TN）乳腺癌。基础TNBC在非洲裔美国人和西班牙裔美国人中更普遍 妇女和没有生物疗法可用于治疗他们。E2启动子结合 E2 Fs是一组受视网膜母细胞瘤（Rb）控制的8种转录因子， 调节几种细胞过程，包括DNA复制、基因组稳定性和凋亡。 广义地说，E2 Fs可以分为转录激活因子（那些促进基因转录的因子）。 转录）和阻遏物（那些停止基因转录）。我在这里提供的初步数据 显示67%的基底/TN乳腺癌过表达E2 F激活剂。已知Rb 通路参与上皮细胞向间质细胞转化（EMT），这是一个导致细胞凋亡的早期过程。 转移;然而，E2 F激活剂驱动EMT在乳腺癌中的作用仍在研究中 调查我们发表的数据表明，E2 F激活因子（E2 F1 -3）的过度表达， 导致有丝分裂调节因子（MPS 1、NEK 2、SGO 1等）的表达增加。 此外，我们最近发表了MPS 1促进EMT和侵袭，NEK 2（一种免疫调节剂）促进EMT和侵袭。 E2 F靶标）在3D培养模型中产生侵入前突起。因此，我们质疑 E2 Fs激活剂的程度失调通过SGO 1促进EMT、侵袭和转移。 具体来说，我们将把我们的研究重点放在E2 F3上，因为它在42%的基底乳腺中过表达。 我们已经证明E2 Fs调节SGO 1蛋白水平的稳定性。我们 中心假设是E2 F3的过表达通过促进肿瘤的发生而促进肿瘤的发生。 EMT通过诱导SGO 1的转录，从而导致侵袭增加， 转移为了检验我们的中心假设，我们将制定以下两个具体目标：（1） 确定E2 F3和SGO 1在转移性、三重性和非转移性肿瘤中的EMT、侵袭和迁移中的作用。 阴性乳腺癌细胞。(2)确定E2 F3基因敲除在MDA-MB-231细胞中的作用 在NOD scid gamma小鼠模型中使用CRISPR/Cas9基因组编辑。我们期望 确定E2 F3过表达如何通过SGO 1促进乳腺EMT机制 癌症，从而有助于迁移、侵袭和转移。如果预期结果是 在这种情况下，小分子可以被设计用于改善当前的治疗，以靶向下游 E2 F3的效应物，因为E2 Fs不能被靶向。