Role of Prdx 1 in Natural Killer Cells and Tumorigenesis

Prdx 1 在自然杀伤细胞和肿瘤发生中的作用

• 批准号: 7118959
• 负责人: CAROLA ANKE NEUMANN
• 金额: $ 10.55万
• 依托单位: MEDICAL UNIVERSITY OF SOUTH CAROLINA
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-02 至 2008-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7118959
• 关键词: aging; antioxidants; athymic mouse; biological signal transduction; calcium flux; chromophore; endocytosis; enzyme linked immunosorbent assay; erythrocytes; fibroblasts; flow cytometry; free radical oxygen; guanine nucleotide binding protein; hydrogen peroxide; immunofluorescence technique; laboratory mouse; leukocyte activation /transformation; natural killer cells; neoplasm /cancer immunology; neoplastic transformation; oxidation reduction reaction; p53 gene /protein; peroxidases; protein localization; tissue /cell culture; transfection /expression vector; tumor necrosis factor alpha; western blottings

DESCRIPTION (provided by applicant) This proposal will help to understand the role of the antioxidant protein peroxiredoxin 1 (Prdx1) in natural killer cell function and tumor susceptibility. Reactive oxygen species (ROS) are one cause of aging and many diseases such as atherosclerosis, autoimmune disorders, neuronal degeneration and cancer. One role of antioxidants is to balance the redox homeostasis to avoid damaging excessive levels of ROS. Prdx1 is also known to be a natural killer (NK) cell activator from red blood cells (RBCs). Loss of Prdx1 in mice leads to premature death due to hemolytic anemia and cancer. Prdx1 deficient mice show decreased NK cell activity and increased cellular damage such as protein oxidation and DNA damage in murine embryonic fibroblasts (MEFs). The Prdx1 deficient mice provide an excellent tool to study the impact of ROS in disease development. Aim 1. To define the role of Prdx1 in natural killer cell activation. It is not fully understood how exogenous Prdx1 activates NK cells. Therefore, it will be demonstrated that Prdx1 is probably secreted by RBCs and co-localizes with the NK cell membrane or cytosol. Localization studies will be done by using fluorescence activated cell sorter (FACS) analysis, immunofluorescence or subcellular fractionation. NK cells will be examined for their ROS content, pH and calcium (Ca2+) content by using fluorescence dyes. Signaling of NK cells will be studied by Western blotting. RBCs and NK cells need Prdx1 to express Tumor Necrosis Factor alpha, I will study whether this production is dependent on Prdx1 antioxidant activity. All studies will include RBCs and NK cells from Prdx1 wildtype and null mice and recombinant Prdx1. Aim 2 will investigate Prdx1 and oxidative stressors as specific regulators of cellular transformation. It is not fully understood whether ROS play a specific role in cellular transformation. MEFs from Prdx1 wildtype and null mice will be chronically stressed with H202 and transformed with oncogenic Ras to test whether ROS can enhance Ras induced-transformation. The tumor suppressor gene p53 is known to be a redox sensitive protein. To study whether p53 is affected by ROS and whether this can lead to enhanced transformation susceptibility by Ras, primary MEFs from p53 and Prdx1 deficient mice will be transformed after chronic stress treatment with H202 and oncogenic Ras.

描述（由申请人提供） 该建议将有助于了解抗氧化剂蛋白过氧蛋白1（PRDX1）在天然杀伤细胞功能和肿瘤敏感性中的作用。活性氧（ROS）是衰老的原因之一，许多疾病，例如动脉粥样硬化，自身免疫性疾病，神经元变性和癌症。抗氧化剂的作用之一是平衡氧化还原稳态，以避免损害过度水平的ROS。 PRDX1也是红细胞（RBC）的天然杀手（NK）细胞活化剂。小鼠中PRDX1的丧失导致由于溶血性贫血和癌症导致过早死亡。 PRDX1缺乏小鼠显示NK细胞活性降低，细胞损伤增加，例如鼠类胚胎成纤维细胞（MEF）中的蛋白质氧化和DNA损伤。 PRDX1缺乏小鼠为研究ROS在疾病发育中的影响提供了一种极好的工具。 目的1。定义PRDX1在天然杀伤细胞激活中的作用。尚未完全了解外源性PRDX1如何激活NK细胞。因此，将证明PRDX1可能由RBC分泌，并与NK细胞膜或细胞质共定位。将使用荧光活化的细胞分析器（FACS）分析，免疫荧光或亚细胞分级分离来完成定位研究。通过使用荧光染料，将检查NK细胞的ROS含量，pH和钙（Ca2+）含量。 NK细胞的信号将通过蛋白质印迹研究。 RBC和NK细胞需要PRDX1来表达肿瘤坏死因子α，我将研究该产量是否取决于PRDX1抗氧化剂活性。所有研究都将包括PRDX1野生型和无效小鼠的RBC和NK细胞以及重组PRDX1。 AIM 2将研究PRDX1和氧化应激源作为细胞转化的特定调节剂。尚未完全了解ROS在细胞转化中是否起特殊的作用。来自PRDX1 WildType和Null小鼠的MEF将长期用H202压力，并用致癌的RAS转化，以测试ROS是否可以增强RAS诱导的转化。已知肿瘤抑制基因p53是氧化还原敏感蛋白。为了研究p53是否受ROS的影响，这是否会导致RAS的转化敏感性增强，在用H202和致癌Ras进行慢性应激治疗后，P53和PRDX1缺乏小鼠的原发性MEF将被转化。