Akoto Theresa, Hadvina Rachel, Jones Skyler, Cai Jingwen, Yu Hongfang, McCord Hayden, Jin Charles X J, Estes Amy J, Gan Lin, Kuo Anthony, Smith Sylvia B, Liu Yutao
Department of Cellular Biology and Anatomy, Medical College of Georgia, Augusta University, Augusta, Georgia, United States.
Center for Biotechnology and Genomic Medicine, Medical College of Georgia, Augusta University, Augusta, Georgia, United States.
Invest Ophthalmol Vis Sci. 2024 Jun 3;65(6):22. doi: 10.1167/iovs.65.6.22.
It is necessary to establish a mouse model of keratoconus (KC) for research and therapy. We aimed to determine corneal phenotypes in 3 Ppip5k2 mouse models.
Central corneal thickness (CCT) was determined using spectral domain optical coherence tomography (SD-OCT) in Ppip5k2+/K^ (n = 41 eyes), Ppip5k2K^/K^ (n = 17 eyes) and 2 knock-in mice, Ppip5k2S419A/+ (n = 54 eyes) and Ppip5k2S419A/S419A (n = 18 eyes), and Ppip5k2D843S/+ (n = 42 eyes) and Ppip5k2D843S/D843S (n = 44 eyes) at 3 and 6 months. Pachymetry maps were generated using the Mouse Corneal Analysis Program (MCAP) to process OCT images. Slit lamp biomicroscopy was used to determine any corneal abnormalities, and, last, hematoxylin and eosin (H&E) staining using corneal sections from these animals was used to examine morphological changes.
CCT significantly decreased from 3 to 6 months in the Ppip5k2+/K^ and Ppip5k2K^/K^ mice compared to their littermate controls. OCT-based pachymetry maps revealed abnormally localized thinning in all three models compared to their wild-type (WT) controls. Slit lamp examinations revealed corneal abnormalities in the form of bullous keratopathy, stromal edema, stromal scarring, deep corneal neovascularization, and opacities in the heterozygous/homozygous mice of the three models in comparison with their controls. Corneal histological abnormalities, such as epithelial thickening and stromal layer damage, were observed in the heterozygous/homozygous mice of the three models in comparison with the WT controls.
We have identified phenotypic and histological changes in the corneas of three mouse lines that could be relevant in the development of animal models of KC.
为了研究和治疗圆锥角膜（KC），建立一种小鼠模型是很有必要的。我们旨在确定3种Ppip5k2小鼠模型的角膜表型。
使用光谱域光学相干断层扫描（SD-OCT）测定Ppip5k2+/K^（n = 41只眼）、Ppip5k2K^/K^（n = 17只眼）以及2种基因敲入小鼠Ppip5k2S419A/+（n = 54只眼）、Ppip5k2S419A/S419A（n = 18只眼）、Ppip5k2D843S/+（n = 42只眼）和Ppip5k2D843S/D843S（n = 44只眼）在3个月和6个月时的中央角膜厚度（CCT）。使用小鼠角膜分析程序（MCAP）生成测厚图来处理OCT图像。使用裂隙灯生物显微镜检查确定是否存在角膜异常，最后，对这些动物的角膜切片进行苏木精和伊红（H&E）染色以检查形态学变化。
与同窝对照相比，Ppip5k2+/K^和Ppip5k2K^/K^小鼠的CCT在3个月至6个月时显著降低。基于OCT的测厚图显示，与野生型（WT）对照相比，所有三种模型均存在异常局限性变薄。裂隙灯检查显示，与对照相比，三种模型的杂合子/纯合子小鼠出现了大疱性角膜病变、基质水肿、基质瘢痕、角膜深层新生血管和混浊等角膜异常。与WT对照相比，在三种模型的杂合子/纯合子小鼠中观察到角膜组织学异常，如上皮增厚和基质层损伤。
我们已经确定了三种小鼠品系角膜中的表型和组织学变化，这些变化可能与KC动物模型的开发相关。
