Exploring novel modulators for rescuing cigarette smoke-induced corneal edema and examining iPSC-derived corneal endothelial cells as a treatment modality

项目摘要 香烟烟雾(CS)是一种复杂的毒素混合物(如尼古丁、一氧化碳) 影响眼睛的多个组织，包括角膜，眼睛的最外面的组织。这个 眼角膜很危险，因为它靠近香烟和飞灰的燃烧端。 角膜内皮是角膜的最内层，负责角膜的水化。 CS导致进行性角膜内皮细胞(CEC)丢失，导致角膜水肿和 视力丧失如果仍然得不到治疗。CS诱导的角膜水肿的严重程度进一步 再加上先前存在的疾病，如角膜内皮营养不良和糖尿病。 我们对蛋白质组谱的初步研究表明，CS触发CEC丢失和 破坏Descemet膜(DM)中的关键结构蛋白。我们进一步确定了 氧化应激相关指标水平升高和CE相关水平下降 香烟烟雾提取物(CSE)对人诱导多能性反应的标志物 干细胞(IPSC)来源的CEC单层培养。同时，我们已经证明，人类 胚胎干细胞(HESC)来源的CECs可在去皮DM(DM)上形成功能性CE 在兔和猴子中没有CE)。此外，我们还证明了注射hESC衍生的 CECs可在无CE和DM的基质上形成功能性CE。 在本应用程序中，我们将在这些数据集的基础上，通过大规模筛选细胞 途径调节剂建立一种非侵入性局部滴眼液方法。同时，我们将 展示了IPSC衍生的CECs注射技术作为一种微创的有效性， 不依赖于供体组织的治疗方式。综合在一起，获得的综合知识 从识别调节器和细胞疗法将通过提供无限的 用于治疗氧化应激引起的CE障碍的来源。

Project Summary Cigarette smoke (CS) is a complex mixture of toxins (e.g., nicotine, carbon monoxide) that affects multiple tissues of the eye including the cornea, the outermost tissue of the eye. The cornea is at high risk due to its proximity to the burning end of the cigarette and flying ash. Corneal endothelium (CE) is the innermost layer of the cornea responsible for corneal hydration. CS results in progressive corneal endothelial cell (CEC) loss leading to corneal edema and vision loss if remains untreated. The severity of CS-induced corneal edema is further compounded with preexisting morbidities such as corneal endothelial dystrophies and diabetes. Our preliminary studies of proteome profiling have demonstrated that CS triggers CEC loss and disruption of critical structural proteins in Descemet’s membrane (DM). We further identified elevated levels of oxidative stress-associated indicators and decreased levels of CE-associated markers in response to cigarette smoke extract (CSE) treatment in human induced pluripotent stem cell (iPSC)-derived CEC monolayer cultures. In parallel, we have shown that human embryonic stem cell (hESC)-derived CECs can form a functional CE on denuded DM (DM without CE) in rabbits and monkeys. Moreover, we have also shown that injected hESC-derived CECs can form a functional CE on denuded stroma (stroma without CE and DM). In this application, we will build on these datasets through large-scale screening of cellular pathway modulators to establish a non-invasive topical eye drop approach. In parallel, we will demonstrate the efficacy of the iPSC-derived CECs injection technique as a minimally invasive, donor tissue-independent, treatment modality. Taken together, the combined knowledge gained from identifying modulators and cell therapy will make a paradigm by offering an unlimited source for the treatment of oxidative stress-induced CE disorders.