Ontogenic changes in erythroid gene expression

红系基因表达的个体发生变化

基本信息

批准号：
6814413
负责人：
MARCUS O MUENCH
金额：
$ 22.73万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2004
资助国家：
美国
起止时间：
2004-09-30 至 2006-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/6814413
关键词：
SDS polyacrylamide gel electrophoresis cell differentiation clinical research erythroid stem cell erythropoiesis flow cytometry gene expression gene expression profiling globin growth /development hematopoiesis human subject microarray technology phage display

项目摘要

DESCRIPTION (provided by applicant): Erythropoiesis dominates hematopoiesis during fetal life to accommodate the rapid expansion in fetal blood volume. Some developmental differences are known to distinguish fetal from adult erythropoiesis such as changes in globin expression and responsiveness to erythropoietin by erythroid progenitors. We hypothesize that a number of unknown genes are differentially regulated between fetal and adult erythroid precursors to accommodate the distinct functions of fetal and adult erythrocytes. Microarray technology is a powerful tool used to compare gene expression between cell populations. To best apply this technology to decipher gene expression during fetal and adult erythropoiesis we have embarked on an effort to improve the tools used to isolate erythroid precursors at different stages of maturation. A panel of single-chain variable fragment antibodies (scFv Abs) was developed, using phage display technology, which recognizes immature nucleated erythrocytes. We aim to determine the molecular identity and expression pattern of the cell-surface markers recognized by these novel reagents. At least three cell surface markers appear to be recognized by the scFv Abs with unique but overlapping expression during erythropoiesis. In addition, to identify the gene products recognized by the scFv Abs we propose to characterize the expression of these cell surface molecules during fetal and adult erythroid development. The second aim of this proposal is to profile gene expression at different stages of fetal and adult erythroid development. This will be accomplished by isolating erythroid precursors at different stages of differentiation from fetal liver and adult bone marrow and then analyzing gene expression using microarray technology. These experiments will both profile the expression of genes during erythroid development and compare expression between adult and fetal erythrocytes. A detailed understanding of gene expression that occurs during the growth and maturation of erythroid progenitors is likely to contribute to improved methods of genetic therapy for hemoglobinopathies.

描述（申请人提供）：胎儿时期红细胞生成主导造血，以适应胎儿血容量的快速扩张。已知一些发育差异可以区分胎儿和成人红细胞生成，例如珠蛋白表达的变化和红细胞祖细胞对红细胞生成素的反应性。我们假设一些未知基因在胎儿和成人红细胞前体之间受到差异性调节，以适应胎儿和成人红细胞的不同功能。微阵列技术是用于比较细胞群之间基因表达的强大工具。为了最好地应用这项技术来破译胎儿和成人红细胞生成过程中的基因表达，我们已着手改进用于在不同成熟阶段分离红细胞前体的工具。使用噬菌体展示技术开发了一组单链可变片段抗体（scFv Abs），该技术可识别未成熟的有核红细胞。我们的目标是确定这些新型试剂识别的细胞表面标记的分子身份和表达模式。至少三个细胞表面标志物似乎被 scFv Ab 识别，在红细胞生成过程中具有独特但重叠的表达。此外，为了鉴定 scFv Ab 识别的基因产物，我们建议表征胎儿和成人红系发育过程中这些细胞表面分子的表达。该提案的第二个目标是分析胎儿和成人红系发育不同阶段的基因表达。这将通过从胎儿肝脏和成人骨髓中分离不同分化阶段的红系前体细胞，然后使用微阵列技术分析基因表达来实现。这些实验将分析红细胞发育过程中基因的表达，并比较成人和胎儿红细胞之间的表达。对红系祖细胞生长和成熟过程中发生的基因表达的详细了解可能有助于改进血红蛋白病的基因治疗方法。