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

探索新型调节剂来挽救香烟烟雾引起的角膜水肿并检查 iPSC 衍生的角膜内皮细胞作为治疗方式

• 批准号: 10723408
• 负责人: Muhammad Ali Riaz
• 金额: $ 9.02万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-30 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10723408
• 关键词: Address; Adherent Culture; Advocate; Affect; Animal Model; Attenuated; Blindness; CRISPR/Cas technology; Carbon Monoxide; Cell Culture Techniques; Cell Density; Cell Line; Cell Therapy; Cells; Cigarette; Clinical; Complex Mixtures; Cornea; Corneal Diseases; Corneal Endothelium; Corneal edema; Critical Pathways; Data; Data Set; Descemet' s membrane; Diabetes Mellitus; Disease; Early treatment; Edema; Endothelial Cells; Eye; Eyedrops; Genetic; Goals; Graft Rejection; Human; Hydration status; In Vitro; Individual; Injection of therapeutic agent; Injections; Keratoplasty; Knowledge; Lead; Mediating; Methodology; Modality; Molecular; Monkeys; Morbidity - disease rate; NQO1 gene; Natural regeneration; Nicotine; Oryctolagus cuniculus; Outcome; Oxidative Stress; Oxidative Stress Induction; Pathway interactions; Patients; Pre-Clinical Model; Proteome; Reporter; Repression; Research; Severities; Signal Pathway; Signal Transduction; Smoking History; Source; Structural Protein; Techniques; Therapeutic; Tissue Donors; Tissues; Topical application; Toxin; Transplantation Surgery; Treatment Efficacy; Validation; anterior chamber; cigarette smoke; cigarette smoke-induced; effective therapy; efficacy evaluation; endothelial dysfunction; experimental study; exposure to cigarette smoke; fly ash; high risk; human embryonic stem cell; in vivo; induced pluripotent stem cell; inhibitor; insight; minimally invasive; novel; novel therapeutics; oxidative damage; pre-clinical; response; restoration; screening; stem cells

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.

项目摘要 香烟烟雾（CS）是一种复杂的毒素混合物（例如，尼古丁、一氧化碳）， 影响眼睛的多个组织，包括角膜，眼睛的最外层组织。的 角膜由于接近香烟的燃烧端和飞灰而处于高风险中。 角膜内皮（CE）是负责角膜水合作用的角膜的最内层。 CS导致进行性角膜内皮细胞（CEC）损失，导致角膜水肿， 如果不治疗，视力会丧失。CS诱导的角膜水肿的严重程度进一步增加。 并伴有先前存在的疾病如角膜内皮营养不良和糖尿病。 我们对蛋白质组分析的初步研究表明，CS触发CEC损失， 破坏后弹力膜（DM）中的关键结构蛋白。我们进一步确认 氧化应激相关指标水平升高，CE相关指标水平降低 人诱导多能干细胞中响应香烟烟雾提取物（CSE）处理的标志物 干细胞（iPSC）衍生的CEC单层培养物。与此同时，我们已经表明，人类 胚胎干细胞（hESC）来源的CEC可以在裸露的DM（DM）上形成功能性CE 不含CE）。此外，我们还表明，注射hESC衍生的 CEC可以在裸露的基质（没有CE和DM的基质）上形成功能性CE。 在这个应用中，我们将通过大规模筛选细胞， 通路调节剂，以建立非侵入性局部滴眼液方法。同时，我们将 证明了iPSC衍生的CEC注射技术作为微创， 不依赖供体组织的治疗方式。综合起来， 从识别调节剂和细胞疗法将通过提供一个无限的模式， 用于治疗氧化应激诱导的CE障碍。