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）： Marcel Bessis在50年前提出了特定的骨髓（BM）微环境促进了谱系血液前体的分化，他在第二个研究中描述了二次研究的结构，该结构是由中央巨噬细胞（M）制成的结构，该结构是由差异化阶段的成年细胞周围的。尽管使用体外重构分析的研究清楚地表明，骨髓衍生的M源在红细胞生成中的作用，但目前尚无体内证据表明M€是必需的，甚至在产后红细胞生成中起作用。从BMM的定义性质较低以及直到最近缺乏特定的遗传模型的定义性质不足的本质性质，这是确定体内知识步骤的主要原因。我们已经定义了BM单核吞噬细胞的子集，并表明CD169+ M =调节造血干细胞小裂。我们的初步研究还表明，BM CD169+ M？促进红细胞生成，因为它们在5-氟尿嘧啶或苯基羟基甘替津诱导的贫血后，其在体内耗竭有选择性的体内耗竭损害了促红细胞生成。此外，在突变的JAK2V617F的转基因过表达的多性性无血病（PV）的遗传模型中，我们发现M deptetion归一化的血细胞分位归一化，这表明PV中的红细胞生成是意外的，这取决于Microenvironment的信号。基于这些初步结果，我们建议在特定目标1中进一步评估BMM的分子基础和功能，我们将在体内分子机制中定义BM红细胞生殖位（红细胞生成岛）的体内分子机制。我们将与M. Narla博士（NYBC）和J. Chasis（Berkeley）合作描述CD169+红细胞岛，并通过流式细胞仪和M. Merad博士（Mt. Sinai）获得有关CentralM的新见解。我们将为先前建议在红细胞岛（VCAM1，ITGAV和MAEA）中发挥作用的候选接收器生成巨噬细胞特异性的遗传模型。在特定的目标2中，我们将研究骨髓m的贡献，我们将分析诸如镰状细胞疾病和thalassya的红细胞性压力模型，后者与S. ​​Rivella博士（Cornell）合作。然后，我们将使用M€Zhao（俄克拉荷马州）和Tony Green博士（剑桥）合作评估巨噬细胞在转基因JAK2V617F模型中的影响，并使用M€使用特定的CD169 -DTR耗竭和M€ - 在第一个目标中产生的特定遗传模型。在特定的目标3中，我们将研究骨髓恢复后促进红细胞生成中心巨噬细胞的潜在治疗益处。我们将与R. Stanley博士，Einstein博士合作使用转基因模型（CSF1过表达）和药物方法（长效IL-4复合物）。这些研究将首次定义BM巨噬细胞在促红细胞生成中的体内作用，并可能导致新的方法来调节以Erythron扩张为特征的疾病中的红细胞生成。 （抽象的结尾）