The role of NF-kappaB signaling in breast cancer metastasis

项目摘要 转移，即癌细胞从原发性肿瘤向继发部位的扩散，是肿瘤转移的主要原因。 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的肿瘤 远处器官的病灶。然而，只有原发性肿瘤内的癌细胞亚群能够在肿瘤细胞中表达。 完成整个转移级联，因为它们必须具有传播能力并含有肿瘤启动因子。 或类似茎的能力。这种致命的细胞群表达高水平的MenaINV，一种促转移亚型， 肌动蛋白调节蛋白Mena的，并能够使用门道的内渗和传播 称为转移的肿瘤微环境（TMEM）位点。初步研究得出的假设是， NF-kB信号通路可能是肿瘤中Mena表达和干性的重要调节因子 细胞，通过与巨噬细胞的相互作用， 与Notch 1信号通路的串扰。这些都是非常复杂和依赖于上下文的信号。因此，我们认为， 这些研究在对活体小鼠的肿瘤进行成像时，将使用生物传感器来确定下游后果。 NF-kB激活对肿瘤细胞中Mena表达和干性的影响， TMEM。我们将使用两种乳腺癌转移模型，其中人或小鼠乳腺癌细胞被转移到乳腺癌细胞中。 注射到小鼠体内并使其生长肿瘤。小鼠表达荧光标记的巨噬细胞， 内皮细胞我们将在肿瘤上植入一个成像窗口， 肿瘤细胞在天然肿瘤微环境中的作用。根据实验，肿瘤细胞将 表达生物传感器以监测NF-kB信号传导、干性和Mena启动子活性的激活。目标1将 确定肿瘤细胞与巨噬细胞碰撞后NF-kB信号的激活是否引起明显的 细胞表型相比，激活NF-κ B信号而不与巨噬细胞碰撞。是否 通过巨噬细胞-肿瘤细胞接触抑制notch信号传导影响NF-κ B信号传导的活化， 得到考验目标2将监测NF-kB、干性和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内渗能力和干细胞样细胞的形成。的 然后用或不用NF-kB抑制剂处理小鼠，以检查NF-kB信号传导是否控制了 干性或Mena表达，以及肿瘤细胞通过TMEM的内渗是否需要NF-κ B信号传导。 破译产生干细胞和内渗肿瘤细胞群的机制- 对我们进一步理解转移至关重要。Condeelis实验室的研究环境 在阿尔伯特·爱因斯坦医学院提供了培训，合作，科学的优秀机会， 讨论和职业发展。本提案中拟定的拟议研究和培训计划将 指导转移，生物传感器开发和大体积高分辨率活体小鼠模型 成像技术，确保掌握建立独立实验室所需的所有工具。

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.