THE UBIQUITIN PATHWAY IN CORNEAL SCARRING

项目摘要：泛素途径在角膜瘢痕形成中的作用 由于损伤、创伤或感染导致的角膜疤痕可导致衰弱的混浊和 永久性视力丧失。急性瘢痕形成，类似于慢性纤维化，以免疫细胞为特征。 被称为肌成纤维细胞的细胞的浸润和持续存在。我们发现去泛素酶 (配音)USP10是角膜疤痕形成途径的关键调节因子。体内敲除USP10导联 能显著减少肌成纤维细胞的发育、细胞的凋亡、CD45+的存在 愈合伤口中的免疫细胞和纤维化的细胞外基质。我们提议测试中央 假设USP10是角膜瘢痕中肌成纤维细胞持久性的关键调节因子。 因子在伤口愈合中的作用。具体地说，由于USP10是αv-整合素和p53在特定情况下的配音 目的1我们将揭示USP10在整合素循环和p53动态中的复杂作用。 人角膜成纤维细胞和小鼠体内。成纤维细胞和成纤维细胞之间的通讯 巨噬细胞在瘢痕形成的结果中起着关键作用。在具体目标2中，我们将开发新的3D和 2.5D水凝胶系统，具有“可调节的硬度”，模拟3D环境，接近于 正在愈合的角膜。利用这些水凝胶，小鼠角膜成纤维细胞将与小鼠骨共同培养。 骨髓来源巨噬细胞(M0/M1/M2)中USP10在巨噬细胞介导中的作用 肌成纤维细胞的发育和收缩。在特定的目标3中，我们使用了三个单独的小鼠模型 将评估USP10在免疫细胞渗入角膜伤口中的作用。

Project Summary: The ubiquitin pathway in corneal scarring Scarring in the cornea resulting from injury, trauma, or infection can lead to debilitating opacities and permanent vision loss. Acute scarring, similar to chronic fibrosis, is characterized by immune cell infiltration and the persistence of cells termed myofibroblasts. We have found that the deubiquitinase (DUB) USP10 is a key regulator of scarring pathways in the cornea. Knockdown of USP10 in vivo leads to a significant reduction in the development of myofibroblasts, cell apoptosis, the presence of CD45+ immune cells, and fibrotic extracellular matrix in a healing wound. We are proposing to test the central hypothesis that USP10 is a key regulator of myofibroblast persistence in a corneal scar via its multi- factorial role in wound healing. Specifically, since USP10 is a DUB for αv-integrins and p53, in Specific Aim 1 we will unravel the complex role of USP10 in integrin recycling and p53 dynamics in primary human corneal fibroblasts and in mice in vivo. The communication between fibroblasts and macrophages plays a critical role in scarring outcomes. In Specific Aim 2 we will develop novel 3D and 2.5D hydrogel systems with “tunable stiffness” that mimic a 3D environment close to the stiffness of the healing cornea. Using these hydrogels, mouse corneal fibroblasts will be cocultured with mouse bone marrow derived macrophages (M0/M1/M2) to elucidate the role of USP10 in macrophage-mediated myofibroblast development and contraction. In Specific Aim 3 using three separate mouse models we will assess the role of USP10 in immune cell infiltration into a corneal wound.