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: 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
• 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

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.

项目总结/摘要 每年约有450，000名女性死于乳腺癌，使其成为乳腺癌最常见的原因 全球女性癌症死亡率最高，其中大多数死亡是由转移性疾病造成的。在 在转移瘤的治疗中，初始药物反应率仅为约50%，相比之下， 治疗原发性肿瘤。此外，对这些全身性治疗的耐药性通常会发展得更多。 在转移性环境中迅速，从而强调迫切需要提高我们对 控制耐药性发展的机制。虽然化疗有效地消除了肿瘤细胞， 它还显示出在细胞和组织中同时诱导各种反调节反应 这导致了保护性环境的发展，促进了少数人的生存 剩余的肿瘤细胞，通常被称为癌症干细胞（CSC）。尽管许多研究都集中在 关于肿瘤细胞耐药的内在机制，很少有人研究外在机制。 由宿主细胞和组织环境的这些变化介导的耐药机制。 骨膜蛋白（Periostin，POSTs）是一种在正常组织发育过程中发现的结构支持蛋白，但其 还显示在癌症发展和转移中表达增加。在转移过程中， POT 3增强肿瘤细胞中促生存Wnt和Akt信号通路以防止应激诱导的肿瘤细胞凋亡。 细胞凋亡和增加CSC的数量，以促进组织的早期转移定植。 重要的是，PO 43的产生主要由TGF-β诱导，TGF-β是一种在肿瘤细胞中上调的分子。 化疗后的肿瘤环境。基于这些观察，我们假设 化疗后产生的过氧化物酶诱导了一个保护性的，有利于生存的环境， 通过激活Wnt和/或Akt信号传导诱导CSC表型而建立转移 途径，并且PO 4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的 关于转移性肿瘤环境中治疗引起的变化的知识差距，揭示了 在转移性环境中，POR 4在介导获得性耐药性中的重要作用。在目标1中，我们 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 人乳腺癌细胞的化学抗性。我们将确定这种耐药性是否通过 使用蛋白质印迹激活Wnt和/或Akt信号传导途径并诱导CSC表型， 流式细胞仪对于aim 2，我们将使用乳腺癌转移的小鼠模型来鉴定关键的细胞因子 以及它们的细胞来源调节化疗后已建立的转移灶内的POR 4产生。 这将通过流式细胞术、荧光显微镜和RT-qPCR完成，并通过在 在小鼠模型中这些细胞因子的体内抗体中和。最后，在目标3中，我们将确定 清除巨噬细胞对已建立的肺转移瘤中药物诱导的过氧化物酶产生的影响 使用转基因小鼠和小分子药物，消耗肿瘤相关的巨噬细胞。充分 阐明了在重新建立化学敏感性时POR 4信号传导中断的可能性，建立了 转移性肿瘤生长对单独化疗或化疗与 将使用生存研究直接比较巨噬细胞消耗或POSTN或TGF-β的中和， 生物发光成像和转移小鼠模型中的组织学。