DISSECTING THE HEMATOPOIETIC GROWTH FACTOR NETWORK

剖析造血生长因子网络

• 批准号: 2142965
• 负责人: GERALD M SEGAL
• 金额: $ 10.22万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-05-15 至 1995-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2142965
• 关键词: antisense nucleic acid; biological signal transduction; bone marrow; cell cycle; colony stimulating factor; fibroblasts; gene complementation; gene expression; gene induction /repression; genetic regulation; hematopoiesis; hematopoietic growth factor; hematopoietic stem cells; interleukin 1; interleukin 6; intracellular; laboratory mouse; messenger RNA; neutralizing antibody; northern blottings; nucleic acid sequence; oligonucleotides; polymerase chain reaction; protooncogene; radioimmunoassay; tissue /cell culture; transcription factor; vascular endothelium; western blottings

Hematopoietic regulation involves an elaborate network of interacting cells and cytokines in which hierarchies of gene expression are complicated by autocrine and paracrine amplification mechanisms. Each cytokine exhibits a variety of biological activities, some of which are synergistic with those of other cytokines. In a system of such complexity, an experimental technique which inhibits the expression of specific genes would greatly facilitate the analysis of questions of function, linkage, and causality. To this end, we have demonstrated that antisense oligodeoxynucleotides complementary to GM-CSF or G-CSF MRNA inhibit expression of the corresponding growth factor in IL-1-induced endothelial cells or fibroblasts in a sequence-specific fashion. In this proposal, we will examine the determinants of the efficiency of inhibition of gene expression by antisense oligonucleotides and the cellular and molecular mechanisms which mediate this inhibition. This approach would be particularly useful in two experimental settings: (1) long-term bone marrow culture (in which it is difficult to deliver antibodies that interrupt juxtacrine mechanisms) and (2) studies on mechanisms of IL-1 signal transduction (these are entirely intracellular and not amenable to manipulation with antibodies). Accordingly, we will employ antisense oligonucleotides to analyze directly the role of GM- CSF, G-CSF, IL-6, and stem cell factor (kit ligand) in the initiation and maintenance of hematopoiesis in long-term bone marrow culture. We will clarify the role of c-fos, c-jun, and c-raf-1 in IL-1-induced hematopoietic growth factor expression using antisense oligonucleotides to specifically inhibit their expression.

造血调节涉及一个复杂的相互作用网络 细胞和细胞因子，其中基因表达的层次是 并伴有自分泌和旁分泌放大机制。每个 细胞因子具有多种生物学活性，其中一些是 与其它细胞因子的那些协同。在这样的系统中， 复杂性，一种抑制表达的实验技术， 特定的基因将极大地促进问题的分析， 功能、联系和因果关系。为此，我们已经证明， 与GM-CSF或G-CSF mRNA互补的反义寡脱氧核苷酸 抑制IL-1诱导的相应生长因子的表达 内皮细胞或成纤维细胞。在这 建议，我们将研究效率的决定因素， 抑制基因表达 通过反义寡核苷酸和 介导这种抑制的细胞和分子机制。这 这种方法在两种实验环境中特别有用：（1） 长期骨髓培养（在这种情况下， 干扰阿曲他克林机制的抗体）和（2）关于 IL-1信号转导机制（这些完全是细胞内的 并且不适于用抗体操作）。因此，我们将 利用反义寡核苷酸直接分析GM- CSF、G-CSF、IL-6和干细胞因子（试剂盒配体） 以及在长期骨髓培养中维持造血。我们 c-fos、c-jun和c-raf-1在IL-1诱导的细胞凋亡中的作用 使用反义寡核苷酸表达造血生长因子 来特异性抑制它们的表达。

项目成果

Repression of Fanconi anemia gene (FACC) expression inhibits growth of hematopoietic progenitor cells.

抑制范可尼贫血基因 (FACC) 的表达会抑制造血祖细胞的生长。

• DOI: 10.1172/jci117405
• 发表时间: 1994
• 期刊: The Journal of clinical investigation
• 作者: Segal,GM;Magenis,RE;Brown,M;Keeble,W;Smith,TD;Heinrich,MC;BagbyJr,GC
• 通讯作者: BagbyJr,GC