Definition of the Microenvironment in Breast Cancer

乳腺癌微环境的定义

基本信息

批准号：
9107039
负责人：
MINA BISSELL
金额：
$ 49.17万
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/9107039
关键词：
Acinus organ component Actins Alternative Splicing Antibodies Architecture Automobile Driving Award Basement membrane Behavior Biochemical Breast Breast Cancer cell line Breast Epithelial Cells Cell Communication Cell Nucleus Cells ChIP-seq Chromatin Communication Data Dystroglycan EGFR inhibition Electron Microscopy Epigenetic Process Equilibrium Extracellular Matrix Extracellular Matrix Proteins Extracellular Space 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 Neoplasms 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 Nuclear Export Nuclear Proteins Pathway interactions Peptide Hydrolases Phenotype Photobleaching Production Protein Isoforms RNA Splicing Regulation Resolution Role Series Signal Pathway Signal Transduction Source Specificity Surface TP53 gene Techniques Testing Therapeutic Tissues Tumor Suppressor Proteins Western Blotting Work base cancer cell deprivation insight malignant breast neoplasm malignant phenotype mammary epithelium mammary gland development novel prevent public health relevance response transcription factor transcriptome sequencing tumor progression unpublished works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（通过应用提供）：核细胞和组织结构的丧失是恶性进展的最早表现之一。为了使组织正常运行，必须保持组织结构。我们假设三十年前假设，细胞外基质（ECM）蛋白和核us之间存在相互的“信号整合计划”。如果这种交流保持平衡，则保持组织稳态。但是，互惠信号中任何步骤的破坏最终都为恶性发展打开了机会。重大发现包括发现层粘连蛋白-11（LN1）的不可或缺作用基底膜（BM）的组成部分。 LN1（例如特定的整联蛋白或多糖蛋白）及其下游介质对LN1的活性或表面受体的功能障碍对LN1的破坏，会破坏整合计划，并导致乳房上皮细胞无法正确偏振并成为静脉曲张，从而导致恶性进展。在先前的提交中，我们定义了与LN1/乳腺细胞相互作用相互作用涉及的许多途径，这些途径培养基的整合，并且在破坏时会导致混乱和恶性肿瘤，如果在3D中进行了纠正，请恢复恶性表型。在上一个融资周期中，我们获得了意外的数据，即通过加速肌动蛋白从细胞核（N-肌动蛋白）加速肌动蛋白来静止，如果预防了肌动蛋白，则细胞将不会输入静止。我们发现LN1激活p53并产生不同的p53同工型。 N-肌动蛋白的出口和p53状态的变化都提供了新颖和未知的途径，将ECM与细胞核的变化联系起来，这导致基因表达改变，这决定了细胞稳态与恶性进展。我们已经组建了一支出色的专家团队，以合作这些研究。我们将使用等源性乳腺癌进展系列，还原性乳腺成形术和基因工程小鼠的细胞来研究BM中的LN1如何调节N-肌动蛋白输出和p53状态。 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由于LN1信号传导。我们提出的工作将提供有关组织如何保持健康的基本知识，并提供针对微环境的潜在新分子和途径，以及在乳腺癌进展过程中可能会出现问题的细胞内信号级联。