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: 8617707
• 负责人: Daniele Marie Gilkes
• 金额: $ 9.06万
• 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 2016-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8617707
• 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; Cancer Etiology; Cancer Patient; Cancer cell line; Cell Line; Cells; Cessation of life; Chemotaxis; Clinic; Clinical Trials; Collagen; Critiques; Data; Deposition; Diagnostic; Environment; Enzymes; Event; Extracellular Matrix; Fibroblasts; Future; Gene Expression; Goals; Hand; Hypoxia; Hypoxia Inducible Factor; In Vitro; Individual; Integrins; Lead; Leadership; Ligands; Lung; Malignant Neoplasms; Mammary Neoplasms; Mentors; Metastatic Lesion; 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; Writing; 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; meetings; migration; novel; novel strategies; overexpression; public health relevance; receptor; treatment strategy; tumor; tumor microenvironment; tumor progression; tumorigenesis

Project Abstract 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.

项目摘要 转移是癌症死亡的首要原因。在所有与肿瘤发生有关的过程中， 局部侵袭和转移的形成是临床上最相关的，但了解最少。 在大多数实体肿瘤中发现的肿瘤内缺氧与发生 转移和治疗失败。癌细胞通过增加生长因子来适应低氧微环境 低氧诱导因子(HIF-1和HIF-2)的活性。驱动HIF的机制- 必须确定受调控的转移，以便确定有效的治疗策略 有可能阻止转移。我们的初步数据显示，HIF的表达促进了胶原蛋白的表达 在体内和体外HIF-1依赖的方式在癌细胞和成纤维细胞中的沉积 胶原、脯氨酸羟基酶和赖氨酸羟基酶的转录上调。我们还发现，胶原蛋白 羟基酶在乳腺癌细胞自发转移到肺中是必不可少的 和小鼠的淋巴组织。另一方面，我们发现低氧诱导了整合素的表达 受体以不依赖于ECM的方式。这些观察结果使我们提出了一个模型，在这个模型中 缺氧诱导因子同时诱导细胞外基质成分(配体)及其整合素(受体)的产生 以增强下游信号事件，从而协同增强转移。建议数 研究人员将通过产生生理性ECM底物来测试这一模型，这些底物概括了 体内细胞外基质的组成。在目标1中，我们将检验HIF-1或HIF-2转录上的假设 调节癌细胞中几个整合素亚基的表达。在指导K99阶段， 我们将确定在低氧条件下整合素基因表达的HIF依赖模式。在.期间 在独立的R00期，HIF对整合素的调节机制将被确定。在目标2中，我们 将检验这一假设，即缺氧诱导和HIF依赖的整合素表达导致 3D培养模型系统中ECM黏附、运动性、侵袭性和基质收缩的增加 新的生物物理检测方法。在指导的K99阶段，将产生细胞系来抵消 低氧对整合素表达的影响，并在所述的检测中进行了测试。在目标3中，我们将测试 假设在低氧条件下产生的细胞外基质上培养乳腺癌细胞将 增强下游整合素信号。在K99阶段，我们将确定整合素 与低氧的细胞外基质相互作用可增强其表达。在R00阶段，我们将测试 假设在缺氧条件下诱导的一些整合素是HIF诱导所必需的 转移。有了这些信息，在R00阶段，我们将系统地评估每个 使用动物模型确定哪些步骤需要整合素(S) 乳腺癌细胞的表达(目标4)。综上所述，我们希望这些数据将导致新的 转移性乳腺癌的治疗策略。 约翰霍普金斯大学独特的环境有许多优势，这些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，塞门扎博士和 共同导师Denis Wirtz博士是各自领域的领导者。他们的领导力和一个 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。