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REGULATION OF TISSUE FACTOR PATHWAY INHIBITOR EXPRESSION

组织因子途径抑制剂表达的调节

基本信息

批准号：
6181932
负责人：
Madhu Satya Bajaj
金额：
$ 10.58万
依托单位：
SAINT LOUIS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
1996
资助国家：
美国
起止时间：
1996-07-01 至 2001-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/6181932
关键词：
anticoagulants antisense nucleic acid cell adhesion fibroblasts gel mobility shift assay gene expression gene induction /repression genetic regulatory element human subject inhibitor /antagonist monocyte northern blottings protease inhibitor thromboplastin transcription factor

项目摘要

The primary goal of these studies is to investigate the regulation of inducible tissue factor pathway inhibitor (TFPI) gene expression in monocytes and fibroblasts. Using Northern blotting analysis and TEPI assays, the applicant had previously demonstrated that endothelium is the primary physiologic site of TFPI synthesis and that monocytes and fibroblasts normally express none to very little TFPI. However, the applicant's preliminary data indicate that TFPI expression is vastly upregulated in adherent monocytes and monocytic U93 cells. In this proposal, the applicant will define the adhesion response elements in the TFPI gene that upregulate TFPI expression in monocytes and U937 cells. The applicant has also found that fibroblasts significantly upregulate TFPI expression in response to serum and in this proposal she plans to define the serum response elements that upregulate its expression in fibroblasts. preliminary data also suggest that adherent monocytes and U937 cells, serum-stimulated fibroblasts and several transformed cell lines that express TFPI also express GATA-2 transcription factor. In gel mobility shift assays, factor(s) from nuclear extracts of HepG2 cells (a transformed cell line that expresses both TFPI and GATA-2 transcription factor) appear to bind to a DNA fragment from the TFPI promoter region containing a putative GATA motif. Additionally, GATA-2 transcription factor antisense oligomers significantly decreased the expression of TFPI and GATA-2 transcription factor mRNA in adherent UJ937 cells. Thus GATA-2 transcription factor may be required for TFPI gene expression. This possibility will be further tested in monocytes and fibroblasts by the use of sense and antisense strategies. Preliminary data was generated under the guidance of the sponsor, Paul Bajaj, who is an expert ina the field of coagulation and thrombosis. The applicant will continue to work in the laboratory of Paul Bajaj for the proposed studies. She will also receive constant guidance from the co- sponsor, Joel Eissenberg who is an experienced and recognized molecular biologist. The applicant will meet frequently with the advisory committee (Drs. Sly, Huang, Hyers, and Payvar) to seek guidance and discuss her progress. Thus ample resources and vast expertise is available to the applicant to perform the proposed studies. It is anticipated that the studies will yield important new information on the regulation of inducible TEPI gene expression during an inflammatory response.

这些研究的主要目标是调查诱导型组织因子途径抑制剂 (TFPI) 基因表达单核细胞和成纤维细胞。使用 Northern 印迹分析和 TEPI 化验，申请人之前已证明内皮细胞是 TFPI 合成的主要生理部位以及单核细胞和成纤维细胞通常不表达或表达很少的 TFPI。然而，申请人的初步数据表明，TFPI表达量极大在贴壁单核细胞和单核 U93 细胞中上调。在这个提案中，申请人将在 TFPI 基因上调单核细胞和 U937 细胞中的 TFPI 表达。这申请人还发现成纤维细胞显着上调 TFPI 对血清的反应表达，在本提案中，她计划定义上调其在成纤维细胞中表达的血清反应元件。初步数据还表明贴壁单核细胞和 U937 细胞，血清刺激的成纤维细胞和几种转化细胞系表达TFPI也表达GATA-2转录因子。凝胶流动性移位分析，来自 HepG2 细胞核提取物的因子（转化的同时表达 TFPI 和 GATA-2 转录因子的细胞系）出现结合 TFPI 启动子区域的 DNA 片段，其中包含假定的 GATA 主题。此外，GATA-2 转录因子反义寡聚物显着降低 TFPI 和 GATA-2 的表达贴壁 UJ937 细胞中的转录因子 mRNA。因此GATA-2 TFPI 基因表达可能需要转录因子。这将通过使用在单核细胞和成纤维细胞中进一步测试可能性有义和反义策略。初步数据是在赞助商 Paul 的指导下生成的 Bajaj是凝血和血栓领域的专家。这申请人将继续在Paul Bajaj的实验室工作拟议的研究。她还将得到同事的持续指导赞助商 Joel Eissenberg 是一位经验丰富且公认的分子生物学家。申请人将经常与顾问委员会会面（Sly 博士、Huang 博士、Hyers 博士和 Payvar 博士）寻求指导并讨论她进步。因此，充足的资源和丰富的专业知识可供申请人进行拟议的研究。预计研究将产生有关诱导调节的重要新信息炎症反应期间 TEPI 基因的表达。