GENE EXPRESSION DURING OSTEOCLAST DIFFERENTIATION

破骨细胞分化过程中的基因表达

• 批准号: 3792168
• 负责人: JOHN M CHIRGWIN
• 金额: --
• 依托单位: UNIVERSITY OF TEXAS HLTH SCIENCE CENTER
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3792168
• 关键词: 25 hydroxycholecalciferol; antisense nucleic acid; calcitonin; calmodulin; carbonate dehydratase; cell differentiation; gene expression; genetic mapping; hormone regulation /control mechanism; hydrogen; membrane transport proteins; myristates; nucleic acid hybridization; nucleic acid probes; osteoclasts; phorbols; physiologic bone resorption; retinoids; superoxide dismutase; tissue /cell culture; transfection; tumor necrosis factor alpha

The osteoclast is a unique cell which arises from a hematopoietic precursor it shares with cells of the granulocyte and monocyte lineages. When activated, it becomes polarized, forms a ruffled border and causes release of bone mineral and proteolytic degradation of the bone matrix. The goal of this project is to elucidate changes in expression of specific genes during osteolysis and during differentiation of the precursors. We plan to quantify changes in the expression of two classes of specific genes: a) those directly associated with osteolysis (such as carbonic anhydrase II, the lysosomal protease cathepsin 0, the ruffled border proton pump, and superoxide dismutase) and b) hormonal responses of these to regulatory factors such as calcitonin, vitamin A, phorbol myristate acetate, 1,25 dihydroxyvitamin D-3 and tumor necrosis factor. We will also plan to determine the functional importance of these gene products by manipulating the phenotype of cultured avian osteoclasts. This will be done by the introduction of DNA constructs which express sense or antisense RNA, thereby raising or lowering the levels of specific cellular marker proteins independent of hormonal regulation of the cells. Two model systems will be used for these studies. The human promyelocytic leukemic cell line HL-60 shares phenotypic features with early precursor cells in the osteoclast lineage. This cell line will be used as a representative of an early osteoclast progenitor. As a model for mature osteoclasts, we will use isolated chicken osteoclasts. These cells, unlike mammalian osteoclasts, are available in sufficient quantities to allow application of the techniques of molecular biology.

破骨细胞是一种独特的细胞，源自造血前体 它与粒细胞和单核细胞谱系的细胞共享。什么时候 激活后，它会极化，形成褶皱边界并导致释放 骨矿物质和骨基质的蛋白水解降解。目标是 该项目旨在阐明特定基因表达的变化 骨溶解和前体细胞分化过程中。我们计划 量化两类特定基因表达的变化：a) 那些与骨质溶解直接相关的酶（如碳酸酐酶 II、 溶酶体蛋白酶组织蛋白酶 0、褶皱边界质子泵，以及 超氧化物歧化酶）和 b）这些对调节的激素反应 降钙素、维生素 A、佛波醇肉豆蔻酸酯醋酸酯等因子，1,25 二羟基维生素 D-3 和肿瘤坏死因子。我们还将计划 通过操作确定这些基因产物的功能重要性 培养的禽类破骨细胞的表型。这将由 引入表达有义或反义RNA的DNA构建体， 从而提高或降低特定细胞标记蛋白的水平 独立于细胞的激素调节。 这些研究将使用两个模型系统。人类早幼粒细胞 白血病细胞系 HL-60 与早期前体细胞具有相同的表型特征 破骨细胞谱系中的细胞。该细胞系将用作 早期破骨细胞祖细胞的代表。作为成熟的典范 破骨细胞，我们将使用分离的鸡破骨细胞。这些细胞不同于 哺乳动物的破骨细胞数量充足，可以 分子生物学技术的应用。