In vivo function of macrophage in healthy and diseased erythropoiesis

巨噬细胞在健康和患病红细胞生成中的体内功能

• 批准号: 8417074
• 负责人: Paul S Frenette
• 金额: $ 43.46万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-26 至 2016-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8417074
• 关键词: Acute; Anemia; Angiopoietin-1; Animals; Blood; Bone Marrow; Bone Marrow Purging; Bone Marrow Transplantation; Breeding; CXCL12 gene; Cells; Chemotherapy-Oncologic Procedure; Collaborations; Commit; Complex; Dichloromethylene Diphosphonate; Disease; Disease model; Erythroblasts; Erythroid; Erythroid Progenitor Cells; Erythropoiesis; Flow Cytometry; Fluorouracil; Generations; Genes; Genetic; Genetic Models; Hematocrit procedure; Hematopoiesis; Hematopoietic stem cells; Hemolysis; Hereditary Disease; Homeostasis; In Vitro; Integrins; Interleukin-4; Island; Knock-in Mouse; Knowledge; Lead; Liposomes; Macrophage Colony-Stimulating Factor; Marrow; Mesenchymal Stem Cells; Modeling; Molecular; Mononuclear; Mus; Mutate; Nature; Oklahoma; Pathogenesis; Phagocytes; Phenylhydrazines; Play; Polycythemia; Polycythemia Vera; Population; Production; Recovery; Role; Seminal; Sickle Cell Anemia; Signal Transduction; Specificity; Spleen; Staging; Stem Cell Factor; Stress; Structure; Testing; Thalassemia; Therapeutic; Time; Transgenic Mice; Transgenic Model; Transgenic Organisms; Vascular Cell Adhesion Molecule-1; abstracting; adhesion receptor; base; chemotherapy; embryonic stem cell; erythroid differentiation; in vivo; insight; irradiation; macrophage; mouse model; nestin protein; novel; overexpression; phenylhydrazine; postnatal; receptor; reconstitution; stem; stem cell niche

DESCRIPTION (provided by applicant): The notion that a specific bone marrow (BM) microenvironment promotes the differentiation of lineage- committed blood precursors was suggested 50 years ago by Marcel Bessis where he described in seminal studies the erythroblastic island, a structure made of a central macrophage (M) surrounded by erythroblasts at varying stages of differentiation. Although studies using in vitro reconstitution analyses have clearly suggested a role for marrow-derived M¿ in erythropoiesis, there is currently no in vivo evidence that M¿ are required or even play a role in postnatal erythropoiesis. A major reason for the deficit of in vivo knowledge stems from the poorly defined nature of BM M¿ and, until recently, the lack of specific genetic models. We have defined subsets of BM mononuclear phagocytes and showed that CD169+ M¿ regulated the hematopoietic stem cell niche. Our preliminary studies also suggest that BM CD169+ M¿ promote erythropoiesis since their selective in vivo depletion compromised erythropoietic recovery after 5-fluorouracil- or phenylhydrazine-induced anemia. Additionally, in a genetic model of polycythemia vera (PV) from transgenic overexpression of mutated JAK2V617F, we have found that M¿ depletion normalized the hematocrit, suggesting that erythropoiesis in PV, unexpectedly, depends on signals from the microenvironment. Based on these preliminary results, we propose to evaluate further the molecular basis and function of BM M¿ in healthy and diseased erythropoiesis. In Specific Aim 1, we will define in vivo molecular mechanisms regulating the BM erythroid niche (erythroblastic island). We will characterize CD169+ erythroblastic islands in collaboration with Drs M. Narla (NYBC) and J. Chasis (Berkeley) and get new insight on central M¿ by flow cytometry and transcriptional profiling collaboration with Dr. M. Merad (Mt. Sinai). We will generate macrophage-specific genetic models for candidate M¿ receptors previously suggested to play a role in the erythroblastic island (Vcam1, Itgav, and Maea). In Specific Aim 2, we will examine the contribution of bone marrow M¿ in chronically diseased erythron. We will analyze models of erythropoietic stress such as sickle cell disease and thalassemia, the latter in collaboration with Dr. S. Rivella (Cornell). We will then evaluate the impact of macrophage in transgenic JAK2V617F models in collaboration with Dr. J. Zhao (Oklahoma) and Tony Green (Cambridge) using M¿-specific CD169-DTR depletion and M¿-specific genetic models generated in the first aim. In Specific Aim 3, we will examine the potential therapeutic benefit of expanding the central macrophage in the erythropoietic recovery after myeloablation. We will use transgenic models (Csf1 overexpression in collaboration with Dr. R. Stanley, Einstein) and pharmacological approaches (long-acting IL-4 complexes). These studies will define for the first time the in vivo roles of BM macrophages in erythropoiesis and will likely lead to novel ways to regulate erythropoiesis in diseases characterized by erythron expansion. (End of Abstract)

描述（由申请人提供）： 50 年前，Marcel Bessis 提出了特定骨髓 (BM) 微环境促进谱系定型血液前体细胞分化的观点，他在开创性研究中描述了成红细胞岛，这是一种由中央巨噬细胞 (M) 组成的结构，周围环绕着巨噬细胞。尽管使用体外重建分析的研究清楚地表明了骨髓来源的 M¿的作用。在红细胞生成过程中，目前没有体内证据表明 M¿体内知识缺乏的一个主要原因是 BM M 的性质不明确。直到最近，我们还定义了 BM 单核吞噬细胞的亚群，并表明 CD169+ M¿我们的初步研究还表明 BM CD169+ M¿促进红细胞生成，因为它们的选择性体内消耗会损害 5-氟尿嘧啶或苯肼诱导的贫血后红细胞生成的恢复。此外，在突变 JAK2V617F 转基因过度表达的真性红细胞增多症 (PV) 遗传模型中，我们发现 M¿耗竭使血细胞比容正常化，这表明 PV 中的红细胞生成出人意料地取决于来自微环境的信号，基于这些初步结果，我们建议进一步评估 BM M 的分子基础和功能。在特定目标 1 中，我们将与 M. Narla 博士 (NYBC) 和 J. Chasis (伯克利) 合作，定义调节 BM 红细胞生态位（红细胞岛）的体内分子机制。 ）并获得对中心 M¿ 的新见解通过与 M. Merad 博士（西奈山）的流式细胞术和转录分析合作，我们将为候选 M¿ 生成巨噬细胞特异性遗传模型。先前认为在红细胞岛中发挥作用的受体（Vcam1、Itgav 和 Maea）在特定目标 2 中，我们将检查骨髓 M¿我们将与 S. Rivella 博士（康奈尔大学）合作分析红细胞生成应激模型，例如镰状细胞病和地中海贫血，然后我们将与 S. Rivella 博士合作评估巨噬细胞对转基因 JAK2V617F 模型的影响。 J.Zhao（俄克拉荷马州）和 Tony Green（剑桥）使用 M¿ -特异性 CD169-DTR 耗尽和 M¿ -第一个目标中生成的特定遗传模型。在特定目标 3 中，我们将研究在清髓后扩大中央巨噬细胞在红细胞生成恢复中的潜在治疗益处。我们将与 R. Stanley 博士合作使用转基因模型（Csf1 过度表达）。 ，爱因斯坦）和药理学方法（长效 IL-4 复合物）这些研究将首次定义 BM 巨噬细胞在红细胞生成中的体内作用，并可能导致。在以红细胞扩张为特征的疾病中调节红细胞生成的新方法。 （摘要完）