The role of the hypoxic ECM on integrin-induced breast cancer metastasis

低氧ECM对整合素诱导的乳腺癌转移的作用

• 批准号: 9148271
• 负责人: Daniele Marie Gilkes
• 金额: $ 24.9万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-02-01 至 2018-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9148271
• 关键词: 2-Oxoglutarate 5-Dioxygenase Procollagen-Lysine; Adhesions; Affect; Animal Model; Award; Behavior; Biochemical; Biological; Biological Assay; Biological Models; Biopsy; Breast Cancer Cell; Breast Cancer cell line; Breast cancer metastasis; Cancer Etiology; Cancer Patient; Cell Line; Cells; Cessation of life; Chemotaxis; Clinic; Clinical Trials; Collagen; Data; Deposition; Diagnostic; Environment; Enzymes; Event; Extracellular Matrix; Fibroblasts; Future; Gene Expression; Goals; Hand; Health; Hypoxia; Hypoxia Inducible Factor; In Vitro; Integrins; Lead; Leadership; Ligands; Lung; Malignant Neoplasms; Mammary Neoplasms; Mentors; Metastatic Lesion; Metastatic breast cancer; Mixed Function Oxygenases; Modeling; Molecular; Mus; Neoplasm Metastasis; Patients; Pattern; Phase; Physiological; Play; Process; Procollagen-Proline Dioxygenase; Production; Regulation; Research; Research Training; Risk; Role; Scientist; Signal Transduction; Solid Neoplasm; Testing; Training; Treatment Failure; Up-Regulation; Work; base; cancer cell; career; career development; cell motility; cell type; clinically relevant; effective therapy; in vitro Model; in vivo; inhibitor/antagonist; insight; lymph nodes; malignant breast neoplasm; migration; novel; novel strategies; overexpression; receptor; treatment strategy; tumor; tumor microenvironment; tumor progression; tumorigenesis

DESCRIPTION (provided by applicant): Metastasis is the leading cause of cancer death. Of all the processes involved in tumorigenesis, local invasion and the formation of metastases are the most clinically relevant, but the least understood. Intratumoral hypoxia, found in the majority of solid tumors, is associated with an increased risk of metastasis and treatment failure. Cancer cells adapt to the hypoxic microenvironment by increasing the activity of the hypoxia-inducible factors (HIF-1 and HIF-2). The mechanisms that drive HIF- regulated metastasis must be determined in order to identify effective treatment strategies with the potential to block metastasis. Our preliminary data showed that HIF expression promotes collagen deposition in vivo and in vitro in a HIF-1? dependent manner in both cancer and fibroblast cells by the transcriptional upregulation of collagen prolyl and lysyl hydroxylases. We also showed that collagen hydroxylase enzymes were essential for the spontaneous metastasis of breast cancer cells to the lung and lymph nodes of mice. On the other hand, we found that hypoxia induced the expression of integrin receptors in an ECM-independent manner. These observations led us to propose a model in which HIFs simultaneously induce the production of ECM components (ligands) and their integrin (receptors) to potentiate downstream signaling events which synergistically enhance metastasis. The proposed research will test this model by generating physiological ECM substrates which recapitulate the composition of ECM in vivo. In aim 1, we will test the hypothesis that HIF-1 or HIF-2 transcriptionally regulates the expression of several integrin subunits in cancer cells. During the mentored K99 phase, we will identify the HIF-dependent pattern of integrin gene expression under hypoxia. During the independent R00 phase, the mechanism of HIF-regulation of integrins will be determined. In aim 2, we will test the hypothesis that hypoxia-induced and HIF-dependent integrin expression causes an increase in ECM adhesion, motility, invasion and matrix contraction in a 3D culture model system using novel biophysical assays. During the mentored K99 phase, cell lines will be generated to counteract the effect of hypoxia on integrin expression and tested in the assays described. In aim 3, we will test the hypothesis that culturing breast cancer cells on ECM produced under hypoxic conditions will potentiate downstream integrin signaling. In the K99 phase, we will determine whether integrin expression is potentiated by interactions with a hypoxic ECM. In the R00 phase, we will test the hypothesis that some of the integrins induced under hypoxic conditions are required for HIF-induced metastasis. With this information in hand, during the R00 phase, we will systematically evaluate each step in the metastatic cascade using animal models to determine which steps require integrin(s) expression by breast cancer cells (aim 4). Taken together, we hope this data will lead to novel strategies for the treatment of metastatic breast cancer. The unique environment at Johns Hopkins has many advantages that will serve to support my training and research plans as well as my future scientific career. My primary mentor, Dr. Semenza and co-mentor, Dr. Denis Wirtz, are leaders in their respective fields. Their leadership together with an intensive career development training plan and the K99 award will facilitate my transition to a successful independent scientist.

描述（由申请人提供）：转移是癌症死亡的主要原因。在所有参与肿瘤发生的过程中，局部侵袭和转移的形成是临床上最相关的，但了解最少。瘤内缺氧，见于大多数 与转移和治疗失败的风险增加有关。癌细胞通过增加缺氧诱导因子（HIF-1和HIF-2）的活性来适应缺氧微环境。必须确定驱动HIF调节转移的机制，以确定具有阻断转移潜力的有效治疗策略。我们的初步数据表明，HIF表达促进胶原沉积在体内和体外的HIF-1？在癌细胞和成纤维细胞中通过胶原蛋白脯氨酰和赖氨酰羟化酶的转录上调以依赖性方式表达。我们还发现，胶原羟化酶是乳腺癌细胞自发转移到小鼠肺和淋巴结所必需的。另一方面，我们发现，缺氧诱导的整合素受体的表达在ECM非依赖性的方式。这些观察结果使我们提出了一种模型，其中HIF同时诱导ECM组分（配体）及其整联蛋白（受体）的产生，以增强协同增强转移的下游信号传导事件。拟议的研究将通过产生生理ECM底物来测试该模型，所述生理ECM底物概括了体内ECM的组成。在目标1中，我们将检验HIF-1或HIF-2转录调节癌细胞中几种整合素亚基表达的假设。在指导K99阶段，我们将确定缺氧条件下整合素基因表达的HIF依赖模式。在独立R 00期，将确定整合素的HIF调节机制。在目标2中，我们将使用新的生物物理测定法在3D培养模型系统中测试缺氧诱导的和HIF依赖性整合素表达导致ECM粘附、运动性、侵袭和基质收缩增加的假设。在指导的K99期，将产生细胞系以抵消缺氧对整联蛋白表达的影响，并在所述测定中进行测试。在目标3中，我们将测试在缺氧条件下产生的ECM上培养乳腺癌细胞将增强下游整合素信号传导的假设。在K99期，我们将确定整合素表达是否通过与缺氧ECM的相互作用而增强。在R 00阶段，我们将测试的假设，在缺氧条件下诱导的一些整合素是HIF诱导的转移所需的。有了这些信息，在R 00阶段，我们将使用动物模型系统地评估转移级联中的每个步骤，以确定哪些步骤需要乳腺癌细胞表达整合素（目的4）。总之，我们希望这些数据将导致转移性乳腺癌治疗的新策略。 约翰霍普金斯独特的环境有许多优势，将有助于支持我的培训和研究计划，以及我未来的科学生涯。我的主要导师Semenza博士和共同导师Denis维尔茨博士都是各自领域的领导者。他们的领导能力，加上密集的职业发展培训计划和K99奖，将促进我向成功的独立科学家的过渡。