Role of ADAMs 9 and 15 in Proliferative Retinopathy

ADAM 9 和 15 在增殖性视网膜病变中的作用

• 批准号: 6946780
• 负责人: Carl Peter Blobel
• 金额: $ 52.31万
• 依托单位: HOSPITAL FOR SPECIAL SURGERY
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-06 至 2009-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6946780
• 关键词: angiogenesis; biological signal transduction; cell cell interaction; enzyme activity; enzyme mechanism; genetically modified animals; laboratory mouse; metalloendopeptidases; proliferative vitreoretinopathy; protein protein interaction; site directed mutagenesis

DESCRIPTION (provided by applicant): Retinal neovascularization is a major cause of blindness, and therefore improvements in our understanding of the molecular mechanism underlying this disease are needed to provide better options for treatment. This proposal focuses on understanding the role of two membrane-anchored metalloproteases, ADAMs (a disintegrin and metalloprotease) 9 and 15, in pathological neovascularization in the retina. We have shown that adam 15-/- mice have strongly decreased pathological neovascularization in a model for retinopathy of prematurity (ROP model), whereas adam 9-/- mice, and adam 9/15-/- double knockout mice have strongly increased pathological neovascularization compared to wildtype controls. We now wish to understand the mechanism underlying the role of ADAMs 9 and 15 in proliferative retinopathy. Specifically, we will: i) Identify whether the metalloprotease domain or other domains of ADAMs 9 or 15 are critical for their role in pathological neovascularization using "knock-in" mice. The metalloprotease domain of ADAMs 9 and 15 may be involved in cleaving substrate proteins, the disintegrin domain/cysteine-rich region may function in cell-cell and cell matrix interactions, and the cytoplasmic domain may have a role in signaling. We will generate "knock-in" mice to evaluate the contribution of different domains to neovascularization. ii) Search for potential substrates or interacting partners of ADAMs 9 and 15 that may be relevant for pathological neovascularization. The most likely hypothesis is that ADAM 9 and ADAM 15 cleave substrate oroteins with a role in angiogenesis. Protein ectodomain shedding is a well-established functional regulator of proteins such as TNFa, EGF-receptor ligands, and Notch and Delta. We will therefore test whether ADAMs 9 or 15, or other ADAMs cleave molecules with a role in angiogenesis, such as VEGFR-2, Tie-1 and 2, PECAM, VE-cadherin, etc. However, should "knock-in" mice demonstrate that regulation of angiogenesis is not due to the metalloprotease activity of ADAM 9 and 15, we will search for functionally relevant extracellular or cytoplasmic interacting proteins. Since ADAMs 9 and 15 are not required for normal development and adult homeostasis, we anticipate that the proposed studies will provide new targets for the design of drugs that regulate pathological neovascularization without affecting normal angiogenesis.

描述（由申请人提供）：视网膜新生血管是致盲的主要原因，因此需要提高我们对这种疾病的分子机制的理解，以提供更好的治疗选择。该提案的重点是了解两个膜锚定的金属蛋白酶，亚当斯（解整合素和金属蛋白酶）9和15，在视网膜中的病理性新生血管形成的作用。我们已经表明，adam 15-/-小鼠在早产儿视网膜病变模型（ROP模型）中具有强烈减少的病理性新血管形成，而adam 9-/-小鼠和adam 9/15-/-双敲除小鼠与野生型对照相比具有强烈增加的病理性新血管形成。我们现在希望了解亚当斯9和15在增殖性视网膜病变中的作用机制。具体而言，我们将： i）使用“敲入”小鼠鉴定亚当斯9或15的金属蛋白酶结构域或其他结构域是否对于它们在病理性新血管形成中的作用是关键的。亚当斯9和15的金属蛋白酶结构域可能参与切割底物蛋白，去整合素结构域/富含半胱氨酸的区域可能在细胞-细胞和细胞基质相互作用中起作用，并且胞质结构域可能在信号传导中起作用。我们将产生“敲入”小鼠来评估不同结构域对新血管形成的贡献。 ii）寻找可能与病理性新血管形成相关的亚当斯9和15的潜在底物或相互作用伴侣。最可能的假设是，ADAM 9和ADAM 15切割底物乳清蛋白，在血管生成中起作用。蛋白质胞外域脱落是诸如TNFa、EGF受体配体以及Notch和Delta的蛋白质的公认的功能调节剂。因此，我们将测试是否亚当斯9或15，或其他亚当斯切割分子与血管生成中的作用，如VEGFR-2，Tie-1和2，PECAM，VE-钙粘蛋白等，然而，应该“敲入”小鼠证明，血管生成的调节是不是由于金属蛋白酶活性的ADAM 9和15，我们将寻找功能相关的细胞外或细胞质相互作用蛋白。 由于亚当斯9和15不是正常发育和成人体内平衡所必需的，我们预计，拟议的研究将提供新的目标，为药物的设计，调节病理性新血管形成，而不影响正常的血管生成。