ANALYSIS OF MECHANISMS OF EARLY HAEMATOPOIETIC STEM CELL DEVELOPMENT CONTROLLED BY FLK1/SCL REGULATORY NETWORK

FLK1/SCL调控网络调控早期造血干细胞发育机制分析

• 批准号: G0900962/1
• 负责人: Alexander Medvinsky
• 金额: $ 158.35万
• 依托单位: University of Edinburgh
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2010
• 资助国家: 英国
• 起止时间: 2010 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=G0900962%2F1
• 关键词: ANALYSIS; MECHANISMS; EARLY; HAEMATOPOIETIC; STEM

The blood system in the animal is maintained through daily generation of billions of different types of erythrocytes, granulocytes and lymphocytes. At the foundation of the blood system lie blood stem cells called haematopoietic stem cells (HSC). Thus, all blood cells in the animal are generated from HSCs. This is the cell type which is responsible for the long-term reconstitution of the blood system of patients who receive bone marrow transplants. Since the number of bone marrow donors is limited, scientists and clinicians are seeking for alternative routes for generation of HSCs in the laboratory conditions. Embryonic stem (ES) cells hold a great promise for regenerative medicine, including production of blood. However, the generation of HSCs from ES cells pose a serious challenge, mainly because many essential details of how the embryo generates HSCs are missing.To a large extent, the progress has been slow since no culture system which would allow us to replicate development of HSCs was available until recently. This research proposal is mainly based on a novel powerful and analytical culture system developed in our laboratory. It allowed us to identify specific stages of development of HSCs in the embryo, at a time when these stages are concealed from scrutiny by traditional methods. We will use this culture system to investigate two genes, called Flk1 and Scl, that are, major regulators of early HSC development. These genes are expressed in various populations of cells of the developing embryo which are potentially involved in development of HSCs. In addition, these genes have a potential to regulate each other?s activity. We will study how Flk1 and Scl genes regulate stages of HSC development by deleting these genes in our culture system using modern recombination technology. Results of such gene ablation will always be verified by a long-term transplantation assay, which is the gold-standard approach for detection of the presence and numbers of HSCs. Additionally, we will carry out more comprehensive analysis of genes expressed at different stages of HSC development and determine how this genetic regulatory network is modified after deletion of Flk1 and Scl. To this end, we will use a modern automated system called Solexa. This approach will allow us to create a computerised genetic network which is involved in early generation of HSCs. In the longer term, this model will be a useful resource for testing the role of other genes in development of HSCs.

动物中的血液系统是通过每天每天产生数十亿种不同类型的红细胞，粒细胞和淋巴细胞来维持的。血液系统的基础是血管干细胞，称为造血干细胞（HSC）。因此，动物中的所有血细胞都是由HSC产生的。这是负责接受骨髓移植的患者血液系统长期重构的细胞类型。由于骨髓供体的数量有限，因此科学家和临床医生正在寻求在实验室条件下产生HSC的替代途径。胚胎干细胞（ES）细胞对再生医学（包括血液的产生）具有巨大的希望。但是，从ES细胞产生的HSC产生构成了一个严重的挑战，主要是因为胚胎如何缺少胚胎的许多基本细节。直到最近才能使用HSC。该研究建议主要基于我们实验室开发的新型强大而分析的培养系统。它使我们能够确定胚胎中HSC的特定发展阶段，而传统方法对这些阶段进行了审查。我们将使用该培养系统来研究两个称为FLK1和SCL的基因，即早期HSC开发的主要调节剂。这些基因在发育中的胚胎的各种细胞中表达，这些细胞可能参与HSC的发展。另外，这些基因有可能相互调节的活性。我们将研究FLK1和SCL基因如何使用现代重组技术在我们的培养系统中删除这些基因来调节HSC开发的阶段。这种基因消融的结果将始终通过长期移植测定法验证，这是检测HSC存在和数量的金标准方法。此外，我们将对在HSC开发的不同阶段表达的基因进行更全面的分析，并确定在删除FLK1和SCL后如何修改该遗传调节网络。为此，我们将使用一个名为Solexa的现代自动化系统。这种方法将使我们能够创建一个与早期HSC相关的计算机化遗传网络。从长远来看，该模型将是测试其他基因在HSC开发中的作用的有用资源。