Extracellular matrix remodeling in ocular anterior segment development

眼前节发育中的细胞外基质重塑

基本信息

批准号：
9189617
负责人：
SUNEEL S APTE
金额：
$ 35.66万
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：
2014
资助国家：
美国
起止时间：
2014-12-01 至 2018-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9189617
关键词：
ADAMTS9 gene Adhesions Adult Affect Alleles Anterior Back Birth Blindness Brain Candidate Disease Gene Cardiac development Cataract Categories Cell Lineage Characteristics Child Childhood Cleaved cell Cleft Palate Complex Congenital Abnormality Cornea Corneal Opacity Crystalline Lens Cultured Cells Data Defect Development Developmental Course Ectoderm Embryo Endothelial Cells Endothelium Extracellular Matrix Eye Eye Abnormalities Eye Development Eye diseases Fibronectins Gene Mutation Genes Glaucoma Glucosyltransferase Goals Human Impairment In Vitro Infant Inherited Investigation Irido-corneo-trabecular dysgenesis Knowledge Krause-Kivlin syndrome LoxP-flanked allele Mass Spectrum Analysis Mediating Mesenchyme Mesoderm Messenger RNA Metalloproteases Modeling Modification Molecular Morphogenesis Mus Mutagenesis Mutate Names Neural Crest Cell Optic Nerve Outcome Pathway interactions Peptide Hydrolases Phenotype Prevention Process Proteins Proteoglycan Proteolysis Public Health Recombinants Retina Role Site Stimulus Structure Surface Ectoderm Syndrome Thrombospondins Variant Vascular Endothelium Vesicle Vision Visual congenital cataract developmental disease fibrillin-2 functional disability knock-down lens lens morphogenesis malformation novel optic cup pressure prevent public health relevance relating to nervous system versican

项目摘要

DESCRIPTION (provided by applicant): Anterior segment dysgenesis (ASD) encompasses a variety of developmental disorders of ocular anterior segment morphogenesis, a complex process involving several cell lineages and their associated extracellular matrix (ECM). The contribution of ECM remodeling to anterior segment morphogenesis has not been the subject of prior investigation. We recently discovered that loss of a single Adamts9 allele (encoding a secreted metalloprotease) resulted in Peters anomaly (corneal leukoma and persistent lens stalk) and lens defects, including cataract. ADAMTS9 is likely a major substrate for the glucosyltransferase B3GALTL, which is mutated in human Peters plus syndrome (PPS). In PPS, extraocular birth defects, including cleft palate and cardiac development anomalies, in which ADAMTS9 had been previously implicated, accompany Peters anomaly. We hypothesize that ADAMTS9 secreted by endothelial cells of the hyaloid vasculature and by the optic cup, is required for ECM turnover and could act non-autonomously on the lens and neural crest cells to regulate anterior segment morphogenesis. ADAMTS9 cleaves the proteoglycan, versican, about which little is known during eye development, despite its role in eye disease (Wagner syndrome). Also, preliminary studies support investigation of the ECM molecules, fibronectin and fibrillin-2 as potential ADAMTS9 substrates. In aim 1 of this proposal, we will comprehensively characterize ADAMTS9 modification by B3GALTL in vitro, and the consequences of preventing this modification in cultured cells. We will define the developmental, cellular and molecular changes underlying ASD resulting from Adamts9 haploinsufficiency. In aim 2, we will use a new floxed Adamts9 allele for conditional inactivation in vascular endothelium (Tie2-Cre), and optic cup (�re), to delineate how ADAMTS9 expressed by these specific lineages drives anterior segment morphogenesis. We will specifically investigate whether ADAMTS9 interacts with and proteolytically processes versican, fibronectin, and fibrillin-2, both in vitro and in the eye. Together, these aims provide a mechanistic continuum from B3GALTL to fundamentals of eye development to molecular actions of ADAMTS9. Relevance to public health: ADAMTS9 is a novel candidate gene for Peters anomaly, PPS or its variants, and other forms of human ASD and/or congenital cataract for which causative gene mutations are unknown. It will provide answers to a hitherto ignored question, i.e. how does ECM dynamics influence early eye development and what are the key proteases that modify the ECM? It will advance fundamental knowledge of ocular and lens morphogenesis that is highly relevant to ASD, childhood glaucoma and cataract, and provides mechanisms pertinent to the formation and possibly, the prevention of adult glaucoma and cataracts.

描述（由适用提供）：前部失去障碍（ASD）包括眼前段形态发生的多种发育障碍，一个复杂的过程涉及多个细胞谱系及其相关的细胞外基质（ECM）。 ECM重塑对前节形态发生的贡献并不是先前研究的主题。我们最近发现，单个ADAMTS9等位基因（编码分泌的金属蛋白酶）的损失导致Peters异常（角膜白细胞和持久性透镜茎）和透镜缺陷，包括CATAACT。 ADAMTS9可能是葡萄糖基转移酶B3GALTL的主要底物，该基材在人类Peters Plus Plus综合征（PPS）中突变。在PPS中，眼外出生缺陷，包括left裂和心脏发育异常，以前涉及Adamts9，伴随着彼得斯异常。假设ECM营业额是由透明脉管系统的内皮细胞和光杯分泌的ADAMTS9，并且可能在透镜和中性rest细胞上非自治作用以调节前节的形态发生。 ADAMTS9裂解了蛋白聚糖，versican，尽管它在眼科疾病（Wagner综合征）中作用，但在眼部发育过程中几乎没有人知道。此外，初步研究支持ECM分子，纤连蛋白和原纤维素-2作为潜在ADAMTS9底物的投资。在本提案的目标1中，我们将在体外通过B3GALTL进行全面表征ADAMTS9的修饰，以及防止在培养细胞中进行这种修饰的后果。我们将定义由ADAMTS9单倍弥倍不足导致的ASD的发育，细胞和分子变化。在AIM 2中，我们将使用新的Floxed Adamts9等位基因在血管内皮（TIE2-CRE）和视杯（RE）中进行有条件灭活，以描绘这些特定谱系表达的ADAMTS9如何驱动前部段的形成。我们将在体外和眼中特别研究ADAMTS9是否与蛋白水解相互作用并通过蛋白水解处理Versican，纤连蛋白和原纤维素-2。这些目标共同提供了从B3GALTL到眼睛发育基础到ADAMTS9的分子作用的机械连续性。与公共卫生相关：ADAMTS9是彼得异常，PPS或其变体以及其他形式的人ASD和/或先天性白内障的新型候选基因，严重基因突变未知。它将为隐藏的忽略问题提供答案，即ECM动态如何影响早期眼睛的发育？修改ECM的关键蛋白是什么？它将促进与ASD，儿童青光眼和白内障高度相关的眼部和晶状体形态发生的基本知识，并提供与形成和可能的机制相关的机制，即预防成人青光眼和白内障。

项目成果

期刊论文数量（3）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

ADAMTS proteins in human disorders.

DOI：
10.1016/j.matbio.2018.06.002
发表时间：
2018-10
期刊：
Matrix biology : journal of the International Society for Matrix Biology
影响因子：
0
作者：
Mead TJ;Apte SS
通讯作者：
Apte SS

A Selective Extracellular Matrix Proteomics Approach Identifies Fibronectin Proteolysis by A Disintegrin-like and Metalloprotease Domain with Thrombospondin Type 1 Motifs (ADAMTS16) and Its Impact on Spheroid Morphogenesis.

选择性细胞外基质蛋白质组学方法通过具有血小板反应蛋白 1 型基序 (ADAMTS16) 的解整合素样和金属蛋白酶结构域 (ADAMTS16) 鉴定纤连蛋白的蛋白水解作用及其对球体形态发生的影响。

DOI：
10.1074/mcp.ra118.000676
发表时间：
2018
期刊：
Molecular & cellular proteomics : MCP
影响因子：
0
作者：
Schnellmann,Rahel;Sack,Ragna;Hess,Daniel;Annis,DouglasS;Mosher,DeaneF;Apte,SuneelS;Chiquet-Ehrismann,Ruth
通讯作者：
Chiquet-Ehrismann,Ruth