REDOX COORDINATED EXTRACELLULAR MATRIX REMODELING IN CANCER

癌症中氧化还原协调的细胞外基质重塑

• 批准号: 7960360
• 负责人: Melanie A Simpson
• 金额: $ 2.81万
• 依托单位: UNIVERSITY OF NEBRASKA LINCOLN
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-08-01 至 2010-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7960360
• 关键词: Anabolism; Antioxidants; Apoptosis; Archives; Biology; Cell Adhesion; Cell Culture Techniques; Cell Death; Cell Proliferation; Cells; Clinical; Collaborations; Collagen; Computer Retrieval of Information on Scientific Projects Database; Deposition; Enzymes; Extracellular Matrix; Frequencies; Funding; Gene Expression; Gene Expression Regulation; Genes; Grant; HIF1A gene; Human; Hyaluronan; Hyaluronidase; Hypoxia; Implant; Inflammation; Inflammatory; Institution; Link; Malignant Neoplasms; Measures; Mediating; Metabolism; Microscopic; Mus; Neoplasm Metastasis; Oxidation-Reduction; Oxidative Stress; Pathway interactions; Phenotype; Pilot Projects; Polymers; Process; Production; Proline; Prostate; Prostatic Neoplasms; Reactive Oxygen Species; Research; Research Personnel; Resources; Signal Pathway; Signal Transduction; Source; Testing; Tissues; United States National Institutes of Health; Universities; cell behavior; cell motility; clinically relevant; collagenase; cytokine; neoplastic cell; novel therapeutics; response; transcription factor; tumor progression; tumorigenesis; two-photon

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Agressive cancer progression is promoted by hypoxic microenvironment and inflammation, conditions that synergize to select for altered survival signals in the tumor cells. Hypoxia may induce cell death that leads to inflammation and/or remodeling of the extracellular matrix, specifically through deposition and processing of extracellular matrix polymer components such as hyaluronan (HA) and collagen. We hypothesize that hypoxia, by altering the redox status of the tumor cell, regulates HA and collagen metabolizing enzymes in an interdependent fashion, and that this link between redox status and matrix remodeling ultimately determines aggressive tumor cell phenotype. In this pilot project, we will test our hypothesis with three specific aims. Aim 1: Determine whether genes involved in HA and collagen metabolism are regulated coordinately in response to oxidative stress imposed by hypoxia and inflammation. We will culture human prostate tumor cells in normoxic and hypoxic conditions in the presence and absence of pro-inflammatory cytokines. We will quantify expression of specific HA synthetic and processing enzymes, as well as enzymes required for proline metabolism, an antioxidant and major component of collagen, in cell culture and in our archived HA-rich and HA-deficient mouse prostate tumors. Dr. DOnald Becker will collaborate with us for these studies. Aim 2: Quantify changes in biosynthesis and turnover of HA and collagen in response to hypoxia and inflammation. We will measure HA accumulation, hyaluronidase activity, collagenase activity and collagen production in tumor cells grown under normoxic and hypoxic conditions as above. Hyaluronidase enzyme functions will be characterized in collaboration with Dr. Joseph Barycki. Cellular redox status will be evaluated by two-photon microscopic quantification of NAD+/NADH ratios and reactive oxygen species (ROS) in collaboration with Dr. Richard Hallworth at Creighton University. Aim 3: Evaluate phenotype of tumor cells with altered HA and collagen metabolism in response to hypoxia and inflammation. We will measure cell proliferation, apoptosis, cell adhesion and motility of tumor cells with manipulated gene expression in hypoxic and normoxic conditions. Specific signaling pathways involved in mediating these responses will be investigated beginning with redox-sensitive transcription factors implicated in motility (CTBP and HIF-1alpha). Defining crosstalk between hypoxia,matrix turnover and tumor cell behavior will elucidate mechanisms by which hypoxia promotes fibrotic tissue damage during tumor progression, a significant clinical problem. Implanting altered tumor cells in mouse prostates and examining colocalization of fibrotic/hypoxic regions with HA and collagen deposition, as well as scoring metastatic frequency, will establish potential clinical relevance. Further delineation of hypoxic gene regulation pathways that mediate cell responses to matrix remodeling will fill in missing links in our understanding of tumorigenesis and metastasis, possibly yielding new therapeutics.

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 恶性癌症进展是由缺氧微环境和炎症促进的，这些条件协同作用以选择肿瘤细胞中改变的存活信号。 缺氧可诱导细胞死亡，导致炎症和/或细胞外基质重塑，特别是通过细胞外基质聚合物组分如透明质酸（HA）和胶原蛋白的沉积和加工。 我们推测，缺氧，通过改变肿瘤细胞的氧化还原状态，调节HA和胶原代谢酶在一个相互依存的方式，氧化还原状态和基质重塑之间的这种联系，最终决定侵略性肿瘤细胞表型。 在这个试点项目中，我们将测试我们的假设与三个具体目标。 目标1：确定参与HA和胶原代谢的基因是否在缺氧和炎症引起的氧化应激反应中协调调节。 我们将培养人前列腺肿瘤细胞在常氧和缺氧条件下的促炎细胞因子的存在和不存在。 我们将定量表达特定的HA合成和加工酶，以及脯氨酸代谢所需的酶，一种抗氧化剂和胶原蛋白的主要成分，在细胞培养和我们存档的HA丰富和HA缺乏的小鼠前列腺肿瘤。 Donald Becker博士将与我们合作进行这些研究。 目的2：量化缺氧和炎症反应中HA和胶原的生物合成和周转的变化。 我们将测量如上所述在常氧和低氧条件下生长的肿瘤细胞中的HA积累、透明质酸酶活性、胶原酶活性和胶原产生。 将与Joseph Barycki博士合作，对Hybronidase酶的功能进行表征。 将与Creighton University的Richard Hallworth博士合作，通过NAD+/NADH比率和活性氧（ROS）的双光子显微镜定量评估细胞氧化还原状态。 目的3：评估具有改变的HA和胶原代谢的肿瘤细胞对缺氧和炎症反应的表型。 我们将测量细胞增殖，凋亡，细胞粘附和运动的肿瘤细胞与操纵基因表达在缺氧和常氧条件下。 参与介导这些反应的特定信号通路将从参与运动的氧化还原敏感性转录因子（CTBP和HIF-1 α）开始研究。 定义缺氧，基质周转和肿瘤细胞行为之间的串扰将阐明缺氧促进肿瘤进展过程中纤维化组织损伤的机制，这是一个重要的临床问题。 将改变的肿瘤细胞植入小鼠前列腺中，并检查纤维化/缺氧区域与HA和胶原沉积的共定位，以及对转移频率进行评分，将建立潜在的临床相关性。 进一步阐明介导细胞对基质重塑的反应的低氧基因调控途径将填补我们对肿瘤发生和转移的理解中缺失的环节，可能产生新的治疗方法。