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

• 批准号: 9926128
• 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
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-07-01 至 2022-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9926128
• 关键词: AKT Signaling Pathway; Apoptosis; Biological Assay; Breast Cancer Cell; Breast Carcinoma; Breast cancer metastasis; Cancer Cell Growth; Cell Survival; Cell physiology; Cells; Cessation of life; 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; 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 Transduction; Site; Source; Stains; Stress; Stromal Cells; Stromal Change; Stromal Neoplasm; Structure; Systemic Therapy; Tissues; Transforming Growth Factor beta; Transgenic Mice; Tumor-associated macrophages; Up-Regulation; WNT Signaling Pathway; Western Blotting; Woman; Work; acquired drug resistance; base; bioluminescence imaging; cancer cell; cancer stem cell; cancer therapy; cell type; chemotherapy; cytokine; drug development; drug sensitivity; experimental study; improved; in vivo; live cell imaging; lung Carcinoma; lung colonization; lung metastatic; macrophage; malignant breast neoplasm; metastatic colorectal; mortality; mouse model; neoplastic; neoplastic cell; neutralizing antibody; periostin; prevent; response; small molecule; therapeutic target; tumor; tumor growth; tumor microenvironment

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.

项目摘要/摘要 每年约有450,000名妇女屈服于乳腺癌，使其成为最常见的原因 全球女性癌症死亡率，大多数这些死亡是由转移性疾病引起的。在 转移的治疗，初始药物缓解率仅约50％，而在90％中观察到 原发性肿瘤的治疗。此外，对这些全身疗法的耐药性通常会发展更多 在转移环境中很快，因此强调了提高我们对 控制耐药性发展的机制。虽然化学疗法有效消除了肿瘤细胞，但 还显示它同时诱导细胞和组织中的各种反调节反应 宿主的of of the of the of的发展环境的发展，促进了很少的生存 剩余的肿瘤细胞，通常称为癌症干细胞（CSC）。 关于耐药性的肿瘤细胞固有机制，对外部的研究很少 宿主细胞和组织环境中的这些变化介导的耐药性机制。 骨膜蛋白（Postn）是在正常组织发育过程中发现的结构支持蛋白，但 表达也已显示在癌症发展和转移中增加。在转移期间， Postn增强了肿瘤细胞中的促生存的Wnt和Akt信号通路，以防止应激诱导 凋亡并增加了组织早期转移性定植的CSC数量。 重要的是，Postn的产生主要由TGF-β诱导，TGF-β是一种受到调节的分子 暴露于化学疗法后的肿瘤环境。基于这些观察，我们假设 对化学疗法的响应后生产引起了受保护的，促生的环境 通过激活Wnt和\或Akt信号的诱导CSC表型建立的转移 途径，该途径主要由肿瘤相关巨噬细胞的TGF-β调节 （tams）。要回答这些问题，我们提出了3个特定目标，我们预计这将填补一个关键 关于治疗诱导的转移性肿瘤环境变化的知识差距，揭示了 邮政在转移性环境中介导获得的耐药性中的重要作用。在AIM 1中，我们将 使用有机肺样培养物鉴定细胞毒性药物，该药物最强烈影响后介导的后介导的细胞毒性药物 人类乳腺癌细胞中的化学抗性。我们将确定是否通过 使用Western blot和 流式细胞仪。对于AIM 2，我们将使用小鼠乳腺癌转移模型来识别关键细胞因子 他们的细胞来源调节化学疗法后已建立的转移酶的后产生。 这将通过流式细胞仪，荧光显微镜和RT-QPCR来实现，并通过 这些细胞因子在小鼠模型中的体内抗体神经化。最后，在AIM 3中，我们将确定 巨噬细胞部署对既定肺转移中药物诱导的后生产的影响 使用转基因小鼠和小分子药物，这些药物复制与肿瘤相关的巨噬细胞。完全 阐明在化学敏感性重新建立中中断邮政信号的潜力，建立 单独对化学疗法的转移性肿瘤生长反应，或结合两种化学疗法 巨噬细胞的部署或神经化后N或TGF-β的神经化将直接使用生存研究直接比较 在转移小鼠模型中的生物发光成像和组织学。