Elucidating the role and regulation of periostin in therapy-induced chemoresistance in metastatic breast cancer

阐明骨膜素在转移性乳腺癌化疗耐药中的作用和调节

基本信息

批准号：
9164338
负责人：
Daniel Patrick Regan
金额：
$ 12.34万
依托单位：
COLORADO STATE UNIVERSITY
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：
2016-07-01 至 2021-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9164338
关键词：
Antibodies Apoptosis Biological Assay Breast Cancer Cell Breast Carcinoma Breast cancer metastasis Cancer Cell Growth Cell physiology Cells Cessation of life Colon Carcinoma Confocal Microscopy Cytotoxic agent Development Disease Disease remission Drug resistance Environment Exposure to Female Fibroblasts Flow Cytometry Fluorescence Microscopy Histology Human Immune Inflammatory Interleukin-13 Interleukin-4 Interruption Investigation Knowledge Lead Link Lung Maintenance Malignant Neoplasms Mammary Neoplasms Mediating Metastatic Neoplasm to the Lung Metastatic breast cancer Molecular Mus Neoplasm Metastasis Normal tissue morphology Organ Pathway interactions Pharmaceutical Preparations Phenotype Play Population Primary Neoplasm Process Production Proteins Recurrence Regulation Resistance Role Signal Induction Signal Pathway Signal Transduction Site Sorting - Cell Movement Source Staining method Stains Stress Stromal Cells Stromal Change Stromal Neoplasm Systemic Therapy Tissues Transforming Growth Factor beta Transgenic Mice Up-Regulation Western Blotting Woman Work abstracting acquired drug resistance base bioluminescence imaging cancer cell cancer stem cell cancer therapy cell type chemotherapy cytokine drug sensitivity improved in vivo live cell imaging lung metastatic macrophage malignant breast neoplasm mortality mouse model neoplastic neoplastic cell periostin prevent research study response small molecule therapeutic target tumor tumor growth tumor microenvironment

项目摘要

Project Summary/Abstract Approximately 450,000 women succumb to breast cancer each year, making it the most common cause of female cancer mortality globally, with the majority of these deaths resulting from metastatic disease. In the treatment of metastases, the initial drug response rate is only about 50%, as compared to 90% observed in the treatment of primary tumors. Furthermore, resistance to these systemic therapies typically develops more quickly in the metastatic setting, thus emphasizing the critical need to improve our understanding of the mechanisms governing development of drug resistance. While chemotherapy effectively eliminates tumor cells, it has also been shown to simultaneously induce various counter-regulatory responses in the cells and tissues of the host, which leads to the development of a protective environment that promotes survival of few remaining tumor cells, often referred to as cancer stem cells (CSCs). Although numerous studies have focused on tumor cell intrinsic mechanisms of drug resistance, there have been very few investigations into extrinsic mechanisms of drug resistance mediated by these changes in the host cells and tissue environment. Periostin (POSTN) is a structural support protein found during normal tissue development, but whose expression has also been shown to be increased in cancer development and metastasis. During metastasis, POSTN enhances the pro-survival Wnt and Akt signaling pathways in tumor cells to prevent stress-induced apoptosis and increase the number of CSCs for promotion of early metastatic colonization of tissues. Importantly, the production of POSTN is predominately induced by TGF-β, a molecule that is up regulated in the tumor environment following exposure to chemotherapy. Based on these observations, we hypothesize that POSTN production in response to chemotherapy induces a protective, pro-survival environment within established metastases through induction of a CSC phenotype via activation of Wnt and\or Akt signaling pathways, and that POSTN production is regulated primarily by TGF-β from tumor-associated macrophages (TAMs). To answer these questions, we propose 3 specific aims, which we anticipate will fill a critical knowledge gap regarding therapy-induced changes within the metastatic tumor environment, revealing a significant role for POSTN in mediating acquired drug resistance in the metastatic setting. In Aim 1, we will use organotypic lung-like cultures to identify the cytotoxic drug(s) that most strongly induce POSTN-mediated chemoresistance in human breast cancer cells. We will determine if this resistance is mediated through activation of the Wnt and\or Akt signaling pathways and induction of a CSC phenotype using western blot and flow cytometry. For aim 2, we will use mouse models of breast cancer metastasis to identify the key cytokines and their cellular sources regulating POSTN production within established metastases following chemotherapy. This will be accomplished via flow cytometry, fluorescence microscopy, and RT-qPCR, and confirmed by in vivo antibody neutralization of these cytokines in the mouse models. Lastly, in aim 3, we will determine the effects of macrophage depletion on drug-induced POSTN production within established pulmonary metastases using transgenic mice and small molecule drugs, which deplete tumor-associated macrophages. To fully elucidate the potential for interruption of POSTN signaling in res-establishment of chemosensitivity, established metastatic tumor growth responses to chemotherapy alone, or chemotherapy in combination with either macrophage depletion or neutralization of POSTN or TGF-β will be directly compared using survival studies, bioluminescence imaging, and histology in mouse models of metastasis.

项目概要/摘要每年约有 450,000 名女性死于乳腺癌，这使其成为导致乳腺癌的最常见原因。全球女性癌症死亡率，其中大部分是由转移性疾病造成的。治疗转移瘤时，初始药物反应率仅为 50% 左右，而在其他研究中观察到的药物反应率为 90% 此外，对这些全身疗法的耐药性通常会产生更多。迅速在转移环境中，从而强调迫切需要提高我们对转移的理解虽然化疗有效地消除了肿瘤细胞，但控制耐药性发展的机制它还被证明可以同时诱导细胞和组织中的各种反调节反应宿主的，这导致了保护性环境的发展，促进了少数物种的生存尽管许多研究都集中在剩余的肿瘤细胞上，但通常称为癌症干细胞（CSC）。关于肿瘤细胞耐药的内在机制，外在机制的研究还很少。由宿主细胞和组织环境的这些变化介导的耐药机制。 Periostin (POSTN) 是在正常组织发育过程中发现的一种结构支持蛋白，但其研究还表明，在癌症发展和转移过程中，表达量会增加。 POSTN 增强肿瘤细胞中促生存的 Wnt 和 Akt 信号通路，以防止应激诱导细胞凋亡并增加 CSC 的数量，以促进组织的早期转移定植。重要的是，POSTN 的产生主要是由 TGF-β 诱导的，TGF-β 是一种在细胞中上调的分子。基于这些观察，我们勇敢地面对化疗后的肿瘤环境。化疗后产生的 POSTN 会在体内诱导一种保护性的、有利于生存的环境通过激活 Wnt 和/或 Akt 信号传导诱导 CSC 表型来建立转移途径，并且 POSTN 的产生主要由来自肿瘤相关巨噬细胞的 TGF-β 调节（TAM）为了回答这些问题，我们提出了 3 个具体目标，我们预计这将填补一个关键问题。关于治疗引起的转移性肿瘤环境变化的知识差距，揭示了在目标 1 中，POSTN 在介导获得性耐药方面发挥着重要作用。使用器官型肺样培养物来鉴定最强烈诱导 POSTN 介导的细胞毒性药物我们将确定这种耐药性是否是通过人乳腺癌细胞介导的。使用蛋白质印迹法激活 Wnt 和/或 Akt 信号通路并诱导 CSC 表型对于目标 2，我们将使用乳腺癌转移的小鼠模型来识别关键的细胞因子。及其细胞来源调节化疗后已建立转移灶内 POSTN 的产生。这将通过流式细胞术、荧光显微镜和 RT-qPCR 来完成，并由 in 证实最后，在目标 3 中，我们将确定小鼠模型中这些细胞因子的体内抗体中和作用。巨噬细胞耗竭对已建立的肺转移瘤内药物诱导的 POSTN 产生的影响使用转基因小鼠和小分子药物来消耗肿瘤相关的巨噬细胞。阐明了 POSTN 信号传导中断在重建化疗敏感性中的潜力，已建立转移性肿瘤生长对单独化疗或化疗联合化疗的反应巨噬细胞消耗或 POSTN 或 TGF-β 的中和将使用生存研究直接比较，生物发光成像和小鼠转移模型的组织学。