The role of NF-kappaB signaling in breast cancer metastasis

NF-κB信号在乳腺癌转移中的作用

• 批准号: 10299624
• 负责人: Camille Duran
• 金额: $ 7.17万
• 依托单位: ALBERT EINSTEIN COLLEGE 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: 2019-12-01 至 2022-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10299624
• 关键词: Actins; Affect; Basement membrane; Biosensor; Blood Circulation; Blood Vessels; Breast Cancer Cell; Breast Cancer Model; Breast cancer metastasis; Cancer Etiology; Cancer Patient; Cell Communication; Cells; Cessation of life; Clinical; Collaborations; Complex; Data; Development; Dissection; Distant; Distant Metastasis; Endothelial Cells; Ensure; Environment; Event; Extravasation; Gene Expression; Gene Silencing; Human; Image; Imaging Techniques; Implant; In Vitro; Label; Lead; Literature; MDA MB 231; Malignant Neoplasms; Mammary gland; Mediating; Medicine; Metastatic breast cancer; Molecular; Monitor; Mus; NF-kappa B; NFKB Signaling Pathway; Neoplasm Metastasis; Organ; Pathway interactions; Patients; Persons; Phenotype; Population; Primary Neoplasm; Process; Protein Isoforms; Receptor Activation; Reporting; Research; Resolution; Role; Secondary to; Signal Pathway; Signal Transduction; Site; System; Techniques; Testing; Time; Tissues; Training; Transgenic Mice; Tumor Cell Migration; Tumor Tissue; United States; Vascular Endothelial Cell; cancer cell; cancer imaging; cancer invasiveness; cancer stem cell; cancer type; career development; college; experimental study; genetic regulatory protein; improved; in vivo; inhibitor; intravital imaging; macrophage; malignant breast neoplasm; migration; mortality; mouse model; multiphoton imaging; multiphoton microscopy; neoplastic cell; notch protein; prevent; promoter; sensor; stem; stem-like cell; stemness; tool; training opportunity; tumor; tumor growth; tumor initiation; tumor microenvironment

Project Summary Metastasis, the dissemination of cancer cells from the primary tumor to secondary sites, is the leading cause of cancer-related mortality. Metastasis is a multistep process, culminating in formation of clinically detectable tumor foci at distant organs. However, only a subpopulation of cancer cells within the primary tumor is capable of completing the entire metastatic cascade, as they must be dissemination-competent and contain tumor-initiating or stemlike capabilities. This deadly population of cells express high levels of MenaINV, a pro-metastatic isoform of the actin-regulatory protein Mena, and are capable of using doorways for intravasation and dissemination called Tumor Microenvironment of Metastasis (TMEM) sites. Preliminary studies lead to the hypothesis that the NF-kB signaling pathway may be a critical regulator of both the expression of Mena and stemness in cancer cells, promoting intravasation through blood vessels, through interactions with macrophages and through crosstalk with the Notch1 signaling pathway. These are very complex and context dependent signals. Therefore, while imaging tumors in live mice, these studies will use biosensors to determine the downstream consequences of NF-kB activation on Mena expression and stemness in tumor cells which are actively intravasating through TMEM. We will use two models of breast cancer metastasis, where human or mouse breast cancer cells are injected into mice and allowed to grow tumors. The mice express fluorescently labelled macrophages and endothelial cells. We will implant an imaging window over the tumor allowing for imaging at single-cell resolution of the tumor cells in the native tumor microenvironment. Depending on the experiment, the tumor cells will express biosensors to monitor activation of NF-kB signaling, stemness, and Mena promoter activity. Aim 1 will determine if activation of NF-kB signaling in tumor cells following collision with macrophages causes a distinct cellular phenotype compared to activation of NF-kB signaling without collision with macrophages. Whether inhibition of notch signaling through macrophage-tumor cell contact affects activation of NF-kB signaling will also be tested. Aim 2 will monitor the timing and order that the NF-kB, stemness, and Mena biosensors are activated to determine which signals promote the formation of the deadly intravasation competent and stem-like cells. The mice will then be treated with or without NF-kB inhibitors to examine if NF-kB signaling controls the activation of stemness or Mena expression, and if NF-kB signaling is required for intravasation of tumor cells through TMEM. Deciphering the mechanisms that produce a population of tumor cells that are both stem and intravasation- competent is critical to further our understanding of metastasis. The research environment in the Condeelis Lab at The Albert Einstein College of Medicine offers outstanding opportunities for training, collaborations, scientific discussions, and career development. The proposed studies and training plan developed in this proposal will instruct on mouse models of metastasis, biosensor development, and large volume high resolution intravital imaging techniques, ensuring mastery of all the tools needed to establish an independent lab.

项目摘要 转移，即癌细胞从原发肿瘤扩散到继发部位，是导致 癌症相关死亡率。转移是一个多步骤的过程，最终形成临床可检测到的肿瘤。 病灶集中在远处的器官。然而，只有原发肿瘤内的一小部分癌细胞能够 完成整个转移级联，因为它们必须具有传播能力，并包含启动肿瘤的内容 或者像茎一样的能力。这种致命的细胞群表达高水平的MenaINV，一种促进转移的亚型 肌动蛋白调节蛋白Mena，并能够利用门口进行血管内和扩散 称为肿瘤转移微环境(TMEM)部位。初步研究得出的假设是， 核因子-kB信号通路可能是肿瘤中MENA表达和干性的关键调节因子 细胞，通过与巨噬细胞的相互作用和通过 与Notch1信号通路的串扰。这些都是非常复杂和与背景相关的信号。因此， 在对活体小鼠的肿瘤进行成像时，这些研究将使用生物传感器来确定下游后果 核因子-kB活化对肿瘤细胞膜蛋白表达和干细胞活性的影响 TMEM。我们将使用两种乳腺癌转移模型，在这两种模型中，人或鼠的乳腺癌细胞 注射到老鼠体内，让它们长出肿瘤。小鼠表达荧光标记的巨噬细胞和 内皮细胞。我们将在肿瘤上植入一个成像窗口，以允许以单细胞分辨率进行成像 在自然的肿瘤微环境中的肿瘤细胞。根据实验结果，肿瘤细胞将 表达生物传感器以监测核因子-kB信号的激活、茎和MENA启动子的活性。目标1将 确定与巨噬细胞碰撞后肿瘤细胞中核因子-kB信号的激活是否会导致明显的 细胞表型与激活核因子-kB信号而不与巨噬细胞碰撞进行比较。是否 通过巨噬细胞-肿瘤细胞接触抑制Noch信号影响核转录因子-kB信号的激活 接受测试。目标2将监测核因子-kB、茎和MENA生物传感器被激活的时间和顺序 以确定哪些信号促进致命的血管内活性干细胞和干细胞的形成。这个 然后，小鼠接受或不使用核因子-kB抑制剂的治疗，以检查核因子-kB信号是否控制着 干细胞或MENA的表达，以及是否需要核因子-kB信号来通过TMEM进入肿瘤细胞。 破译产生大量既是干细胞又是血管内癌细胞的机制-- 能力是加深我们对转移的理解的关键。康迪利斯实验室的研究环境 在阿尔伯特·爱因斯坦医学院提供了出色的培训、合作、科学 讨论和职业发展。本提案中制定的拟议研究和培训计划将 指导小鼠肿瘤转移模型、生物传感器的研制和大容量高分辨率活体实验 成像技术，确保掌握建立独立实验室所需的所有工具。