Axolotl Hematopoiesis: A Regeneration Model

蝾螈造血：再生模型

• 批准号: 8915413
• 负责人: Edward W Scott
• 金额: $ 33.75万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-07-01 至 2019-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8915413
• 关键词: Acomys; Active Sites; Adult; African; Ambystoma; Ambystoma mexicanum; Amphibia; Animals; Antibodies; Benign; Biological Metamorphosis; Biological Models; Biology; Blood; Blood Cells; Brain; Breeding; Cancer Biology; Carcinogens; Cartilage; Cell Transplants; Cell surface; Cells; Chimera organism; Cicatrix; Colony-Forming Units Assay; Color; Comparative Study; Competence; Complex; Cytomegalovirus; Development; Embryo; Engraftment; Equilibrium; Genetic; Genetic Models; Goals; Grant; Growth; Healed; Heart; Hematopoiesis; Hematopoietic; Hematopoietic System; Homing; Imagery; Immune; Immune system; Immunity; Inflammation; Jaw; Knowledge; Leukocyte Elastase; Leukocytes; Limb structure; Liver; MYB gene; Malignant Neoplasms; Mammals; Maps; Methodology; Microsurgery; Modeling; Morphology; Muramidase; Mus; Muscle; Mutagens; Myelogenous; Myelopoiesis; Natural regeneration; Neoplasms; Organ; Paper; Pathway interactions; Patients; Peroxidases; Play; Proto-Oncogene Proteins c-myb; PubMed; Publishing; Rana; Reagent; Regulation; Relative (related person); Reporter; Reptiles; Research; Role; S100A8 gene; Salamander; Site; Skin; Skin Tissue; Spinal Cord; Spleen; Structure; Tail; Techniques; Testing; Time; Tissues; Transgenic Organisms; Vertebrates; Wound Healing; Zebrafish; carcinogenesis; comparative; design; healing; immune clearance; improved; in vivo; limb regeneration; macrophage; mouse model; mutant; neutrophil; paracrine; progenitor; promoter; public health relevance; reconstitution; regenerative; segregation; tissue regeneration; trait; tumorigenesis

 DESCRIPTION (provided by applicant): In mammals, the ability to regenerate limbs and organs is progressively lost during ontogeny and correlates closely with maturation of immune competence. Research in scar-free wound healing, primarily observed in model systems with dysfunctional neutrophils and macrophages, has led to the hypothesis that the immune system dictates the balance between scarring and regeneration. We seek to test this hypothesis by studying a regenerative animal. Among vertebrates, urodele amphibians, such as the axolotl (Ambystoma mexicanum), display a unique and extensive ability for adult regeneration. Axolotls can replace a wide variety of tissues and complex structures including muscle, cartilage, skin, spinal cord, brain, heart, jaw, and limbs. Axolotl also do not get cancer. Mutagens do not induce transformation in axolotls at worst they induce spurious regeneration or a small benign neoplasia. Enhanced immune clearance of damaged tissue by axolotl leukocytes is another hypothesis we hope to test. To do so, we must better define the axolotl hematopoietic system - particularly the myeloid lineages where very little beyond morphology is known. Defining the genetic pathways required in the hematopoietic system for regeneration via a comparative approach contrasting matched murine and axolotl wound healing/regeneration models is one long-term goal that will be facilitated by these studies. These studies will also facilitate definig the role of hematopoietic cells in axolotl cancer immunity by similar comparative approaches replicating well establishes murine models of oncogenesis in axolotl. This proposal seeks to provide the initial characterization of axolotl hematopoietic ontogeny with a focus on myeloid development. A PubMed search for axolotl HSC, macrophages, or neutrophils yields fewer than 25 relevant papers published since 1972. Therefore, the design of this proposal is more empirical using tested methodology from mammalian and zebrafish studies to delineate axolotl hematopoiesis/myelopoiesis. Heavy emphasis will be placed on identifying/generating antibody reagents for hematopoietic specific cell surface markers. In addition, we will use previously successful hematopoietic specific transgenic reporter constructs (Vav-1, gata-1, PU.1 (made myself)), c-myb, Lyz (lysozyme), MRP8 (mac specific), Mpx (myeloperoxidase) and NE (neutrophil elastase) regulatory/promoter constructs) to generate matching axolotl transgenic strains to facilitate comparative studies. We have two Specific Aims: Aim 1. Defining the Axolotl HSC: Ontogeny, Function and Niche. Aim2. Characterization of Macrophages and Neutrophils in Axolotl.

 描述（申请人提供）：在哺乳动物中，肢体和器官再生能力在个体发育过程中逐渐丧失，并与免疫能力的成熟密切相关。无瘢痕伤口愈合的研究，主要是在具有功能失调的中性粒细胞和巨噬细胞的模型系统中观察到的，已经导致了免疫系统决定瘢痕形成和再生之间的平衡的假设。我们试图通过研究再生动物来验证这一假设。在脊椎动物中，有尾目两栖动物，如蝾螈（Ambystoma mexicanum），显示出独特而广泛的成体再生能力。蝾螈可以取代各种各样的组织和复杂的结构，包括肌肉，软骨，皮肤，脊髓，大脑，心脏，下巴和四肢。蝾螈也不会得癌症。突变剂不会诱导蝾螈发生转化，最坏的情况是，它们会诱导假再生或小的良性瘤形成。美西蝾螈白细胞对受损组织的增强免疫清除是我们希望检验的另一个假设。要做到这一点，我们必须更好地定义美西蝾螈的造血系统-特别是骨髓谱系，在那里除了形态学之外知之甚少。通过对比匹配的小鼠和美西蝾螈伤口愈合/再生模型的比较方法来定义造血系统再生所需的遗传途径是这些研究将促进的一个长期目标。这些研究也将有助于通过类似的比较方法确定造血细胞在蝾螈癌症免疫中的作用，复制并建立蝾螈肿瘤发生的小鼠模型。该建议旨在提供美西螈造血个体发育的初步表征，重点是骨髓发育。PubMed检索美西螈HSC、巨噬细胞或中性粒细胞，自1972年以来发表的相关论文不到25篇。因此，本提案的设计更符合经验，使用来自哺乳动物和斑马鱼研究的测试方法来描述美西螈造血/骨髓生成。重点将放在识别/生成造血特异性细胞表面标志物的抗体试剂上。此外，我们将使用先前成功的造血特异性转基因报告构建体（Vav-1、加塔-1、PU. 1（自己制造））、c-myb、Lyz（溶菌酶）、MRP 8（mac特异性）、Mpx（髓过氧化物酶）和NE（中性粒细胞弹性蛋白酶）调节/启动子构建体）来产生匹配的美西螈转基因株以促进比较研究。我们有两个具体目标：目标1。定义蝾螈HSC：个体发育，功能和生态位。目标2。蝾螈巨噬细胞和中性粒细胞的特征。