Definition of Microenvironment in Breast Cancer

乳腺癌微环境的定义

• 批准号: 6943643
• 负责人: MINA BISSELL
• 金额: $ 34万
• 依托单位: UNIVERSITY OF CALIF-LAWRENC BERKELEY LAB
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-06-01 至 2009-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6943643
• 关键词: DNA methylation; acetylation; acinar cell; basement membrane; biological signal transduction; blocking antibody; breast neoplasms; cell adhesion; cell differentiation; cell line; cellular oncology; cellular polarity; chromosome deletion; clinical research; fibroblasts; gene expression; genetic library; genetic screening; genetically modified animals; human tissue; intercellular connection; laboratory mouse; laminin; mixed tissue /cell culture; myoepithelial cell; neoplastic transformation; transcription factor

DESCRIPTION (provided by applicant): One of the earliest manifestations of malignant progression is loss of tissue organization. This proposal examines the hypothesis that structural integrity is crucial for maintenance of normal breast function and suppression of neoplasia. We propose that characterization of the "signaling integration plans" which establish and sustain acini will drive the discovery of therapeutically-relevant markers of malignancy. Based on our progress in the previous funding period, we now postulate that myoepithelial cells (myoeps) provide crucial structural and polarity cues to luminal cells that enable formation of normal acini, cues likely responsible for myoep tumor suppressor functions. In the last funding period, we replicated acini in "designer" 3D microenvironments and established double-layered acini from purified human myoeps and luminal cells. We immortalized and characterized several breast cell lines and showed that, in culture, luminal cells acquire myoep-specific survival markers (e.g. a6b4 integrin), permitting acinus formation in the absence of myoeps. Importantly, we also showed that myoep production of laminin 1 (Ln-1) is crucial in establishing acinar polarity, as tumor myoeps lacking Ln-1 fail to induce polarity. We expand upon these findings in 4 specific aims: 1-Probe the nature of the "cross-talk" and formation of basement membrane between luminal cells and myoeps using mouse/human chimeras of primary cells or immortalized cell lines, and mouse/mouse chimeras from wild-type and genetically engineered mice. We will assess the roles of heterotypic cell-cell adhesion and morphogens in the cross talk; 2-Explore the genetic and epigenetic mechanisms behind Ln-1 silencing in tumor myoeps (deletions, rearrangements, methylation or acetylation); 3-Identify Ln-1 domains and Ln-1 receptors required for induction of polarity (using Ln-1 fragments, inhibitory peptides, blocking antibodies and cre-lox ablation of receptors); 4-Screen for genes and pathways affecting acinar polarity. Collectively, these experiments address the role of myoeps and the importance and dominance of polar acinar structure in breast function. As loss of appropriate integration of these signals could lead to malignancy, our results may yield new markers and therapeutic strategies to limit or reverse breast tumor progression.

描述（由申请人提供）：恶性进展的最早表现之一是组织组织的丧失。 该提议研究了结构完整性对于维持正常乳腺功能和抑制肿瘤至关重要的假设。 我们建议建立和维持ACINI的“信号整合计划”的表征将推动与治疗性的恶性标记发现。 基于我们在上一个资金期间的进展，我们现在假设肌上皮细胞（MyOEEP）为腔内细胞提供了至关重要的结构和极性线索，从而使肌肉发达形成正常，这可能导致可能导致肌ep肿瘤抑制器功能。 在最后一个资金期间，我们在“设计师” 3D微环境中复制了ACINI，并从纯化的人肌和腔内细胞中建立了双层acini。 我们永生并表征了几种乳腺细胞系，并表明在培养物中，腔细胞获得了肌发行特异性的生存标记（例如A6B4整合素），允许在没有肌eps的情况下形成腺泡。 重要的是，我们还表明，层粘连蛋白1（LN-1）的产生对于建立腺泡极性至关重要，因为缺乏LN-1的肿瘤肌无力无法诱导极性。 我们在4个特定目的中扩展了这些发现：1探针，使用原代细胞或永生细胞系的小鼠/人嵌合体以及野生型和基因工程小鼠的小鼠/小鼠嵌合体使用小鼠/人类嵌合体之间的“交叉对话”的性质以及腔细胞和Myoep之间地下膜形成的性质。 我们将评估异型细胞 - 细胞粘附和形态植物在横式演讲中的作用； 2-探索肿瘤肌E的LN-1沉默背后的遗传和表观遗传机制（缺失，重排，甲基化或乙酰化）；诱导极性所需的3识别LN-1域和LN-1受体（使用LN-1片段，抑制性肽，阻断受体的抗体和Cre-Lox消融）； 4屏，用于影响腺泡极性的基因和途径。 总的来说，这些实验探讨了肌无端的作用以及极性粉刺结构在乳房功能中的重要性和优势。 由于这些信号的适当整合可能会导致恶性肿瘤，因此我们的结果可能会产生新的标记和治疗策略，以限制或逆转乳腺肿瘤的进展。