Regulation of Hematopoiesis

造血调节

• 批准号: 7152943
• 负责人: BEVERLY J. TOROK-STORB
• 金额: $ 41.01万
• 依托单位: FRED HUTCHINSON CANCER RESEARCH CENTER
• 依托单位国家: 美国
• 财政年份: 1984
• 资助国家: 美国
• 起止时间: 1984-08-01 至 2008-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7152943
• 关键词: Abnormal Monocyte; Address; Affect; Aspirate substance; Biopsy; Bone Morphogenetic Proteins; CD34 gene; Candidate Disease Gene; Cell Communication; Cell Line; Cells; Coculture Techniques; Cultured Cells; Data; Detection; Development; Disease Progression; Dysmyelopoietic Syndromes; Dysplasia; Engraftment; Event; Fibroblasts; Gene Expression; Genes; Goals; Growth; Hematopoiesis; Hematopoietic; Human; Immune; In Vitro; Interleukin-1; Knowledge; Laboratories; Lead; Ligands; Maintenance; Marrow; Mediating; Methods; Microarray Analysis; Molecular Profiling; Outcome; PPBP gene; Patients; Population; Process; Proteins; Purpose; Regulation; Research; Research Personnel; Role; Stem cell transplant; Stem cells; Stromal Cells; Therapeutic Intervention; Transplantation; Western Blotting; Work; base; cDNA Arrays; cadherin-6; cell type; comparative; conditioning; cytochemistry; disease characteristic; graft failure; improved; in vitro Assay; in vitro Model; loss of function; macrophage; monocyte; notch protein; osteopontin; progenitor; programs

DESCRIPTION (provided by applicant): The overall goal of this project is to dissect the marrow microenvironment (ME) for the purpose of identifying gene products that work in concert to regulate early events in hematopoiesis. Functionally distinct stromal cell lines have been generated to address this goal: one line supports immature progenitors (HS-27a) and one line drives differentiation (HS-5). Microarray technology has been used to generate gene expression profiles from these lines to identify differentially expressed genes (Data from this comparative analysis is available at htpp://parma.fhcrc.org/M Iwata). In Aim 1, the array data will be confirmed for select genes by Northern and Western blotting and immune cytochemistry. Once their relevance has been established by detection in primary marrow cultures and marrow biopsies, the role of candidate gene products will be investigated by determining how gain and/or loss of function effects the ability of the ME to support hematopoiesis. Aim 2 will address how monocytes/macrophages, which are an integral part of the ME, effect stromal function. Gene expression profiles have been generated for both normal monocytes and for HS-27a cells cultured alone. These have been compared to profiles from the two cell types cultured together to identify significant changes in both monocyte-derived and stroma-derived gene products. Several gene products with significantly altered expression and potential relevance to ME function have been selected for functional studies. These include: osteopontin (OPN) which down regulates Notch on CD34 cells; CXCL7 which may influence fibroblast growth; bone morphogenetic protein antagonist -1(BMPA-1); Cadherin-6 (CDH6) which is expressed by stroma and CD34 cells and mediates homophilic, yet heterotypic cell-cell interactions; and stromal derived factor-1 (SDF-1). The knowledge gained from Aims 1 and 2 will determine how normal monocytes interact with stromal cells and contribute to ME function. This information will be used as a platform to determine in Aim 3 how abnormal monocytes may compromise the ME. Specifically, we hypothesize that abnormal monocytes in patients with myelodysplastic syndrome (MDS) adversely affect the function of an intrinsically normal stroma, giving rise to the dysplasias characteristic of this disease. Additionally, we propose that MDS monocytes, through their retention after nonmyeloablative conditioning, cause the marrow graft failure and consequent disease progression seen in a significant proportion of MDS patients given stem cell transplants. This research plan is based on historical as well as new preliminary studies indicating that monocytes, normal or abnormal, interact with stromal cells to modulate ME function. Hypothetically this can advantageously or adversely influence hematopoiesis in general and stem cell engraftment in particular. A better understanding of these effects should facilitate the development of new therapies to correct dysplasias and optimize stem cell transplantation.

描述（由申请人提供）：该项目的总体目标是剖析骨髓微环境（ME），以识别协同调节造血早期事件的基因产物。为了实现这一目标，已经产生了功能不同的基质细胞系：一种细胞系支持未成熟祖细胞（HS-27a），另一种细胞系驱动分化（HS-5）。微阵列技术已用于从这些品系中生成基因表达谱，以鉴定差异表达的基因（来自该比较分析的数据可在 htpp://parma.fhcrc.org/M Iwata 获得）。在目标 1 中，将通过 Northern 和 Western 印迹以及免疫细胞化学来确认所选基因的阵列数据。一旦通过原代骨髓培养物和骨髓活检中的检测确定了它们的相关性，将通过确定功能的获得和/或丧失如何影响 ME 支持造血的能力来研究候选基因产物的作用。目标 2 将解决 ME 的组成部分单核细胞/巨噬细胞如何影响基质功能。已生成正常单核细胞和单独培养的 HS-27a 细胞的基因表达谱。将它们与一起培养的两种细胞类型的概况进行比较，以确定单核细胞来源和基质来源的基因产物的显着变化。几种表达显着改变且与 ME 功能潜在相关的基因产物已被选择用于功能研究。其中包括： 骨桥蛋白 (OPN)，可下调 CD34 细胞上的 Notch； CXCL7可能影响成纤维细胞生长；骨形态发生蛋白拮抗剂-1(BMPA-1); Cadherin-6 (CDH6) 由基质细胞和 CD34 细胞表达，介导同质但异型的细胞间相互作用；和基质衍生因子 1 (SDF-1)。从目标 1 和 2 中获得的知识将确定正常单核细胞如何与基质细胞相互作用并促进 ME 功能。该信息将用作目标 3 中确定异常单核细胞如何损害 ME 的平台。具体来说，我们假设骨髓增生异常综合征（MDS）患者的异常单核细胞会对本质正常基质的功能产生不利影响，从而导致这种疾病的发育不良特征。此外，我们认为，MDS 单核细胞在非清髓性调理后保留，会导致骨髓移植失败以及随后的疾病进展，这在接受干细胞移植的相当大比例的 MDS 患者中可见。该研究计划基于历史和新的初步研究，表明正常或异常的单核细胞与基质细胞相互作用以调节 ME 功能。假设这通常可以有利或不利地影响造血作用，特别是干细胞植入。更好地了解这些影响应该有助于开发新疗法来纠正发育不良和优化干细胞移植。