Endothelial protease activity, organogenesis and birth defects

内皮蛋白酶活性、器官发生和出生缺陷

基本信息

批准号：
8177298
负责人：
SUNEEL S APTE
金额：
$ 7.85万
依托单位：
CLEVELAND CLINIC LERNER COM-CWRU
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-09-01 至 2013-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8177298
关键词：
ADAMTS9 gene Adult Affect Alleles Antibodies Area Blood Vessels Blood capillaries CD31 Antigens Candidate Disease Gene Cardiac Cells Cleft Palate Congenital Abnormality Congenital Heart Defects Cornea Data Defect Deposition Development Developmental Biology Developmental Process Differentiation and Growth Embryo Embryonic Development Endocardium Endothelial Cells Endothelium Extracellular Matrix Failure Generations Goals Heart Heart failure Histocompatibility Testing Histologic Human In Situ Invaded Knowledge Left Limb Development Mediating Mesenchyme Metalloproteases Modification Morphogenesis Mus Myocardial Myocardium Organ Organogenesis Palate Peptide Hydrolases Phenotype Pregnancy Process Proteins Proteoglycan Proteolysis Regulation Research Right-On Role Secondary Palate Staging Staining method Stains Structure Syndrome Tamoxifen Tumor Angiogenesis Vascular Endothelium Vascularization Work angiogenesis base behavior influence body system cadherin 5 capillary craniofacial endothelial-specific sialomucin gastrulation improved insight mouse development mouse model palatogenesis versican

项目摘要

DESCRIPTION (provided by applicant): The goal of this project is to elucidate the role of vascular endothelium in embryonic development and organogenesis. Microvascular endothelial cells (mEC), which line capillaries, have a well-known function in regulation of angiogenesis, but less is known about their role in organogenesis. Our preliminary data suggests that the evolutionarily conserved, secreted metalloprotease ADAMTS9, a product of microvascular endothelium, has an important role in normal organogenesis. Adamts9-/- mice die by 7.5 days of gestation (E7.5), prior to the onset of angiogenesis and organogenesis. Nevertheless, using mice lacking one allele of Adamts9, we have shown that ADAMTS9 influences both vascular and non-vascular developmental processes. Specifically, Adamts9 expressed by mEC worked cooperatively with Adamts20 expressed by craniofacial mesenchyme in closure of the mouse secondary palate, and also influenced development of the myocardium and angiogenesis. Moreover, ADAMTS9 proteolysis of versican, a large, developmentally critical proteoglycan, was crucial in both the palate and the heart. Since ADAMTS9 is expressed globally by mEC during organogenesis and because versican is widely expressed during embryogenesis, our intriguing observations suggest a broader impact of mEC-produced ADAMTS9 during development. Based on these findings, we hypothesize that through the proteolytic modification of secreted proteins, ADAMTS9 produced by mEC influences surrounding cells during organogenesis. To gain a complete understanding of the developmental impact of mEC-derived ADAMTS9, and to elucidate the underlying mechanisms, a conditional targeting approach is required. The specific aim of this RO3 proposal is to achieve conditional deletion of ADAMTS9 in mEC, and perform morphologic analysis of the resulting mouse phenotype, thereby resolving the overall impact on angiogenesis and organogenesis. This will lay the groundwork for continued work on the identified developmental defects, including their underlying mechanisms. Approach: We will delete Adamts9 specifically in mEC at the earliest developmental stages (E8.5) using a Tie2-Cre strain. In addition, we will undertake regulated conditional deletion in mEC at later developmental stages (E10.5 and later) using tamoxifen-induced Cre expression (via a VE-cadherin-Cre ERT2 mouse). Morphologic analysis of the resulting mouse phenotypes will identify effects on angiogenesis and organogenesis. Significance: In addition to the expected impact on angiogenesis, palatogenesis and myocardial development, it is foreseen that the proposed study will elucidate a widespread role for mEC-expressed ADAMTS9 in organogenesis. The proposed work has immediate significance for common human birth defects, such as cleft palate and cardiac anomalies, and may provide new mouse models for other developmental defects. PUBLIC HEALTH RELEVANCE: ADAMTS9 expressed in endothelial cells influences angiogenesis, palate closure and myocardial maturation during mouse development, and is implicated in cleft palate and heart defects, which are both common, medically significant birth defects. The proposed research is expected to provide fundamental new insights relevant to the role of ADAMTS9 in defective angiogenesis, cleft palate and myocardial non-compaction. The new information obtained will provide new knowledge regarding the functional relationship between vascular endothelium and embryonic development, and could potentially define new directions for treatment of specific birth defects. !

描述（由申请人提供）：该项目的目的是阐明血管内皮在胚胎发育和器官发生中的作用。线毛细血管的微血管内皮细胞（MEC）在调节血管生成方面具有众所周知的功能，但对它们在器官发生中的作用知之甚少。我们的初步数据表明，微血管内皮的产物是进化保守的，分泌的金属蛋白酶ADAMTS9在正常器官发生中具有重要作用。在血管生成和器官发生之前，Adamts9 - / - 小鼠妊娠7.5天（E7.5）死亡。然而，使用缺乏ADAMTS9等位基因的小鼠，我们已经表明ADAMTS9会影响血管和非血管发育过程。具体而言，由MEC表达的ADAMTS9与颅面间质表达的ADAMTS20合作合作，闭合小鼠次生口感，还影响了心肌和血管生成的发育。此外，ADAMTS9的蛋白水解是一种大型，发育症的大型蛋白聚糖，在味觉和心脏上都是至关重要的。由于ADAMTS9在器官发生过程中由MEC全球表达，并且由于在胚胎发生过程中广泛表达了versican，因此我们有趣的观察结果表明，在发育过程中，MEC生产的ADAMTS9对更广泛的影响。基于这些发现，我们假设通过对分泌蛋白的蛋白水解修饰，由MEC产生的ADAMTS9在器官发生过程中会影响细胞周围的细胞。为了完全了解MEC衍生的ADAMTS9的发育影响，并阐明了基本机制，需要采用条件靶向方法。该RO3建议的具体目的是实现MEC中ADAMTS9的条件缺失，并对所得小鼠表型进行形态学分析，从而解决对血管生成和器官发生的总体影响。这将为确定的发育缺陷（包括其潜在机制）继续工作奠定基础。方法：我们将使用TIE2-CRE菌株在最早的发育阶段（E8.5）中特别在MEC中删除ADAMTS9。此外，我们将使用他莫昔芬诱导的CRE表达（通过VE-Cadherin-Cre ERT2小鼠）在后来的发育阶段（E10.5及以后）在MEC中进行调节的条件缺失。对所得小鼠表型的形态分析将确定对血管生成和器官发生的影响。意义：除了预期对血管生成，古质发生和心肌发育的影响外，还可以预见，拟议的研究还将阐明MEC表达的ADAMTS9在器官发生中的广泛作用。拟议的工作对于常见的人类先天缺陷（例如left裂和心脏异常）具有直接的意义，并可能为其他发育缺陷提供新的小鼠模型。公共卫生相关性：在内皮细胞中表达的ADAMTS9影响小鼠发育过程中的血管生成，口感闭合和心肌成熟，并与left裂和心脏缺陷有关，这两者都是常见的医学上重要的先天出生缺陷。拟议的研究有望提供与ADAMTS9在缺陷的血管生成，left裂和心肌无复数中的作用相关的基本新见解。获得的新信息将提供有关血管内皮和胚胎发育之间功能关系的新知识，并有可能定义用于治疗特定出生缺陷的新方向。呢