Extracellular matrix remodeling in ocular anterior segment development

眼前节发育中的细胞外基质重塑

• 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
• 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

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（由申请人提供）：眼前段发育不良（ASD）包括多种眼前段形态发生发育障碍，这是一个涉及多种细胞系及其相关细胞外基质（ECM）的复杂过程。ECM重塑对前段形态发生的贡献尚未被研究。我们最近发现，一个Adamts9等位基因（编码一种分泌的金属蛋白酶）的缺失会导致彼得斯异常（角膜白血病和持续的晶状体柄）和晶状体缺陷，包括白内障。ADAMTS9可能是葡萄糖基转移酶B3GALTL的主要底物，该酶在人类彼得斯综合征（PPS）中发生突变。在PPS中，眼外出生缺陷，包括腭裂和心脏发育异常，包括ADAMTS9，伴随彼得斯异常。我们推测，由玻璃状血管内皮细胞和视杯分泌的ADAMTS9是ECM转换所必需的，并且可以非自主地作用于晶状体和神经嵴细胞以调节前段形态发生。ADAMTS9可切割蛋白聚糖，versican，尽管它在眼病（瓦格纳综合征）中起作用，但在眼睛发育过程中对其知之甚少。此外，初步研究支持ECM分子、纤维连接蛋白和纤维蛋白-2作为潜在的ADAMTS9底物的调查。在本提案的目的1中，我们将全面表征B3GALTL在体外修饰ADAMTS9，以及在培养细胞中阻止这种修饰的后果。我们将定义由Adamts9单倍不全导致的ASD的发育、细胞和分子变化。在目标2中，我们将使用一个新的在血管内皮（Tie2-Cre）和视神经杯（Tie2-Cre）中有条件失活的Adamts9等位基因，来描述这些特定谱系表达的Adamts9是如何驱动前段形态发生的。我们将在体外和眼内研究ADAMTS9是否与纤连蛋白、纤维连接蛋白和纤连蛋白2相互作用和蛋白水解过程。总之，这些目标提供了一个从B3GALTL到眼睛发育基础再到ADAMTS9分子作用的机制连续体。与公共卫生相关：ADAMTS9是Peters异常、PPS或其变体以及其他形式的人类ASD和/或先天性白内障的新候选基因，其致病基因突变未知。它将为迄今为止被忽视的问题提供答案，即ECM动力学如何影响早期眼睛发育以及修饰ECM的关键蛋白酶是什么？它将推进与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础知识，并提供与成人青光眼和白内障的形成和可能的预防相关的机制。

项目成果

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
• 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
• 作者: Schnellmann,Rahel;Sack,Ragna;Hess,Daniel;Annis,DouglasS;Mosher,DeaneF;Apte,SuneelS;Chiquet-Ehrismann,Ruth
• 通讯作者: Chiquet-Ehrismann,Ruth