Role of ADAMs 9 and 15 in Proliferative Retinopathy

描述(申请人提供)：视网膜新生血管是导致失明的一个主要原因，因此需要提高我们对这种疾病潜在的分子机制的了解，以便为治疗提供更好的选择。这项建议侧重于了解两种膜锚定的金属蛋白水解酶ADAMS(去整合素和金属蛋白水解酶)9和15在视网膜病理性新生血管中的作用。我们已经证明，在早产儿视网膜病变模型(ROP模型)中，ADAM 15-/-小鼠显著减少了病理性新生血管，而与野生型对照相比，ADAM 9-/-小鼠和ADAM 9/15-/-双基因敲除小鼠显著增加了病理性新生血管。现在，我们希望了解ADAMS 9和15在增殖性视网膜病变中的作用机制。具体来说，我们会： I)利用“敲入”小鼠，确定ADAMS 9或15的金属蛋白酶域或其他结构域是否对它们在病理性新生血管中的作用起关键作用。ADAMS 9和15的金属蛋白酶结构域可能参与底物蛋白的切割，去整合素结构域/富含半胱氨酸的区域可能参与细胞与细胞和细胞基质的相互作用，细胞质结构域可能参与信号转导。我们将产生“敲入”小鼠，以评估不同结构域对新生血管的贡献。 Ii)寻找可能与病理性新生血管相关的亚当斯9和15的潜在底物或相互作用伙伴。最有可能的假设是，亚当9和亚当15切割底物oroteins，在血管生成中发挥作用。蛋白质胞外结构域脱落是一种成熟的蛋白质功能调节因子，如TNFa、EGF受体配体、Notch和Delta。因此，我们将测试Adams 9或15或其他Adams是否切割在血管生成中起作用的分子，如VEGFR-2、Tie-1和2、PECAM、VE-cadherin等。然而，如果“敲入”小鼠证明血管生成的调节不是由于ADAM 9和15的金属蛋白酶活性，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ADAMS 9和15不是正常发育和成人动态平衡所必需的，我们预计拟议的研究将为设计在不影响正常血管生成的情况下调节病理性新生血管的药物提供新的靶点。

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.