Souverein Erik A, Oak Sanyukta, Fouladian Zachary, Reid Mark W, Fernandez G Esteban, Salas Andrew, Aparicio Jennifer G, Nagiel Aaron, Gnedeva Ksenia
Keck School of Medicine of USC, University of Southern California, Los Angeles, California, United States.
USC Caruso Department of Otolaryngology - Head and Neck Surgery, Keck School of Medicine of USC, University of Southern California, Los Angeles, California, United States.
Invest Ophthalmol Vis Sci. 2025 Jun 2;66(6):69. doi: 10.1167/iovs.66.6.69.
To investigate whether Hippo pathway inhibition via a novel Lats kinase inhibitor (LKI) induces regeneration of retinal pigment epithelium (RPE).
An in vitro scratch "injury" on confluent RPE monolayers was produced using the Woundmaker Tool. A 6×6 grid of laser spots was delivered to the central retinae of Dutch Belted rabbits via indirect laser. Following laser injury, eyes received 50 µL intravitreal LKI or vehicle. Fundus photography and optical coherence tomography (OCT) were performed prior to sacrifice for immunohistological analyses.
In vitro RPE demonstrated accelerated wound healing 24 to 96 hours after injury with LKI versus control treatment (P < 0.0001). In the in vivo rabbit model of retinal degeneration, laser spots in LKI-injected eyes demonstrated hyperpigmentation and RPE layer thickening with shadowing on OCT beginning at 1 week after treatment. There was a marked increase in RPE cross-sectional area (P < 0.0001) and percentage of Ki67+ RPE and Müller glia (P < 0.001), specifically at the laser spots. At 4 weeks, the increase in RPE cross-sectional area persisted (P < 0.001), but Ki67+ cells were no longer observed within the RPE (P = 0.36) or Müller glia (P = 0.48). The response was limited to the damaged regions, and there was no proliferation seen in the intact retina in either control or LKI-treated eyes.
Hippo pathway inhibition via a novel LKI promotes wound healing and RPE proliferation in RPE monolayers in vitro. In a rabbit model of geographic atrophy, LKI treatment facilitated robust RPE regeneration and Müller glia proliferation specifically at the sites of injury.
研究通过新型大肿瘤抑制激酶（Lats）抑制剂（LKI）抑制Hippo信号通路是否能诱导视网膜色素上皮（RPE）再生。
使用伤口制作工具在汇合的RPE单层细胞上造成体外划痕“损伤”。通过间接激光向荷兰带兔的中央视网膜发射6×6网格的激光点。激光损伤后，眼睛接受50 μL玻璃体内注射LKI或赋形剂。在处死前进行眼底照相和光学相干断层扫描（OCT）以进行免疫组织学分析。
与对照处理相比，体外RPE在损伤后24至96小时用LKI处理显示伤口愈合加速（P < 0.0001）。在视网膜变性的体内兔模型中，注射LKI的眼睛中的激光点在治疗后1周开始在OCT上显示色素沉着过度和RPE层增厚并伴有阴影。RPE横截面积显著增加（P < 0.0001），Ki67 + RPE和Müller胶质细胞的百分比显著增加（P < 0.001），特别是在激光点处。在4周时，RPE横截面积的增加持续存在（P < 0.001），但在RPE（P = 0.36）或Müller胶质细胞（P = 0.48）中不再观察到Ki67 +细胞。反应仅限于受损区域，在对照或LKI处理的眼睛的完整视网膜中均未观察到增殖。
通过新型LKI抑制Hippo信号通路可促进体外RPE单层细胞的伤口愈合和RPE增殖。在地图样萎缩的兔模型中，LKI治疗促进了强大的RPE再生和Müller胶质细胞增殖，特别是在损伤部位。
