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Definition of the Microenvironment in Breast Cancer

乳腺癌微环境的定义

基本信息

批准号：
9906592
负责人：
MINA BISSELL
金额：
$ 14.57万
依托单位：
UNIVERSITY OF CALIF-LAWRENC BERKELEY LAB
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-08-01 至 2021-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9906592
关键词：
3-Dimensional Acinus organ component Actins Alternative Splicing Antibodies Architecture Automobile Driving Basement membrane Biochemical Breast Breast Cancer Cell Breast Epithelial Cells Cancer cell line Cell Communication Cell Nucleus Cells ChIP-seq Chromatin Communication Data Dimensions Dystroglycan EGFR inhibition Electron Microscopy Epigenetic Process Equilibrium Extracellular Matrix Extracellular Matrix Proteins Extracellular Space First Independent Research Support and Transition Awards Functional disorder Funding Gel Gene Chips Gene Expression Genetically Engineered Mouse Genotype Glucose Goals Grant Growth Homeostasis Indium-111 Integrins Investigation Knowledge Laminin Laminin Receptor Lead Link Malignant - descriptor Malignant Neoplasms Mammaplasty Mammary Gland Parenchyma Mammary gland Manuscripts Mediating Mediator of activation protein MicroRNAs Microscopy Modeling Modification Molecular Morphogenesis Morphologic artifacts Myoepithelial cell Nitric Oxide Non-Malignant Normal tissue morphology Nuclear Pathway interactions Peptide Hydrolases Phenotype Photobleaching Production Protein Isoforms Proteins RNA Splicing Regulation Resolution Role Series Signal Pathway Signal Transduction Source Specificity Structure Surface TP53 gene Techniques Testing Therapeutic Tissues Tumor Suppressor Proteins Western Blotting Work breast cancer progression cancer cell cell behavior deprivation genetically modified cells insight malignant breast neoplasm malignant phenotype mammary epithelium mammary gland development novel prevent public health relevance response transcription factor transcriptome transcriptome sequencing unpublished works

项目摘要

DESCRIPTION (provided by applicant): Loss of nuclear- cellular- and tissue- architecture is one of the earliest manifestations of malignant progression. In order for a tissue to function properly, tissue architecture must be maintained. We postulated 3 decades ago that there is a reciprocal `signaling integration plan' between the extracellular matrix (ECM) proteins and the nucleus. If this communication remains balanced, tissue homeostasis is maintained. However disruption of any step in reciprocal signaling eventually opens an opportunity for malignant progression. Significant findings include discovery of the indispensible role of laminin-111 (Ln1), a component of the basement membrane (BM). Destruction of Ln1 by unscheduled proteinase activity or dysfunction of surface receptors for Ln1 (such as specific integrins or dystroglycan) and their downstream mediators, disrupts integration plan and leads to inability of breast epithelial cells to correctly polarize and become quiescent thus leading to malignant progression. In previous submission we have defined a number of pathways involved in Ln1/breast cell interactions that mediate integration and, when disrupted, lead to disorganization and malignancy and, if corrected in 3D, revert the malignant phenotype. During last funding cycle we obtained unexpected data that Ln1 signals for quiescence by accelerating the export of actin from the nucleus (N-actin), and if this is prevented, the cells will not enter quiescence. We found Ln1 activates p53 and production of different p53 isoforms. Both the export of N-actin and changes in p53 status provide novel and uncharted pathways linking ECM to changes in the nucleus which lead to altered gene expression that dictate cellular homeostasis vs malignant progression. We have assembled a stellar team of experts to collaborate on these studies. We will use an isogenic breast cancer progression series, primary breast cells from reduction mammoplasty, and cells from genetically engineered mice to investigate how Ln1 in the BM modulates N-actin export and p53 status. We will use techniques such as RNAseq, Mass Spec, ChIPseq, super-resolution microscopy with photobleaching approaches, along with more routine cell, molecular and biochemical techniques such as western blots, IP, IF using additional well-characterized antibodies from our collaborators to investigate changes in gene expression, epigenetic modifications, and changes in p53 isoforms responsible for dramatic alterations in phenotypes as a result of Ln1 signaling. Our proposed work will provide fundamental knowledge about how tissues can remain healthy, as well as providing potential new molecules and pathways that target the microenvironment and the resulting intracellular signaling cascades that can go awry during breast cancer progression.

描述（由申请人提供）：核细胞和组织结构的丧失是恶性进展的最早表现之一。为了使组织正常发挥功能，必须维持组织结构。30年前，我们假设细胞外基质（ECM）蛋白和细胞核之间存在相互的“信号整合计划”。如果这种交流保持平衡，组织的稳态就得以维持。然而，相互信号传导中的任何步骤的中断最终为恶性进展打开了机会。重要的发现包括发现层粘连蛋白-111（Ln1）的不可或缺的作用，基底膜（BM）的一个组成部分。Ln1被非程序性蛋白酶活性或Ln1表面受体（如特异性整联蛋白或肌营养不良蛋白聚糖）及其下游介质功能障碍破坏，破坏整合计划，导致乳腺上皮细胞无法正确生长和静止，从而导致恶性进展。在之前的提交中，我们已经定义了一些参与Ln 1/乳腺细胞相互作用的途径，这些途径介导整合，当被破坏时，导致组织解体和恶性肿瘤，如果在3D中得到纠正，则会逆转恶性表型。在上一个资助周期中，我们获得了意想不到的数据，即Ln1通过加速肌动蛋白（N-actin）从细胞核的输出来发出静止信号，如果阻止这一点，细胞将不会进入静止状态。我们发现Ln1激活p53并产生不同的p53亚型。N-肌动蛋白的输出和p53状态的变化都提供了将ECM与细胞核中的变化连接起来的新的和未知的途径，其导致改变的基因表达，所述基因表达决定细胞稳态与恶性进展。我们已经组建了一个一流的专家团队来合作进行这些研究。我们将使用一个同基因乳腺癌进展系列，原代乳腺细胞从减少乳房成形术，和细胞从基因工程小鼠来研究如何在BM中的Ln 1调节N-肌动蛋白输出和p53状态。我们将使用RNAseq、质谱、ChIPseq、超分辨率显微镜和光漂白方法等技术，沿着更常规的细胞、分子和生物化学技术，如蛋白质印迹、IP、IF，使用我们合作者提供的其他表征良好的抗体来研究基因表达、表观遗传修饰、和p53同种型的变化，作为Ln1信号传导的结果，负责表型的显著改变。我们提出的工作将提供有关组织如何保持健康的基础知识，并提供潜在的新分子和途径，靶向微环境和由此产生的细胞内信号级联，这些信号级联在乳腺癌进展过程中可能出错。