Development of stem cell-based therapies for limbal stem cell deficiency

项目概要/摘要： 角膜盲是世界上第四大致盲原因。角膜缘干细胞缺乏症（LSCD） 由于角膜上皮干细胞或角膜缘干细胞的功能障碍或缺乏而导致的致盲性眼病 角膜基质干细胞（LSC），维持透明角膜上皮的正常稳态。LSCD，通常 在许多常见的眼科疾病中，都可以看到未诊断和最具挑战性的治疗。最理想的治疗 自体LSC移植治疗严重/完全单侧LSCD。移植的有效性 自1997年以来，在超过37项研究中报道了培养的LSC（cLSC）。然而，种植 方案和临床结果测量因研究而异。只有三项研究使用符合GMP的 cLSC。此外，在过去二十年中，成功率没有提高。为了提高疗效， cLSC的安全性特征，使用标准化LSC培养的临床试验和临床方案， 必要为了实现这一目标，我们的实验室建立了一个无外源性和无饲养层的LSC 培养方法，可持续产生具有满足高水平临床应用标准的特性的cLSC。 成功已经开发了可量化的体外参数来表征cLSC，并且新开发了一种新的 体内成像技术可用于量化患者中LSC功能。在这份提案中，我们将建立 这些可量化的体外参数作为cLSC生产的过程中质量控制， 体内参数作为临床结果指标，以客观评估临床结果。初步临床研究 评价GMP条件下生产的cLSC的安全性和有效性。执行 这些体外和体内可量化的参数将使得有可能建立标准化的患者- 特异性干细胞治疗从拟议的研究中学到的知识将构成 开发下一代患者特异性角膜缘干细胞疗法。

Project summary/Abstract: Corneal blindness is the fourth leading cause of blindness in the world. Limbal stem cell deficiency (LSCD) is a blinding eye disorder due to the dysfunction or absence of corneal epithelial stem cells or limbal stem cells (LSCs), which maintain the normal homeostasis of the transparent corneal epithelium. LSCD, which is often undiagnosed and most challenging to treat is seen in many common eye diseases. The most desired treatment is transplantation of autologous LSCs in severe/total unilateral LSCD. The efficacy of transplantation of cultivated LSCs (cLSCs) has been reported in more than 37 studies since 1997. However, the cultivation protocol and the clinical outcome measures vary from study to study. Only three studies used GMP-compliant cLSCs. In addition, the success rate has not improved over the last two decades. To improve the efficacy and safety profile of the cLSCs, clinical trials using a standardized LSC cultivation and clinical protocol are necessary. To accomplish this goal, our laboratory has established a xenobiotic-free and feeder-free LSC culture method that can consistently produce cLSCs with properties that meet the criteria for high-level clinical success. Quantifiable in vitro parameters have been developed to characterize cLSCs and a newly developed in vivo imaging technique can be used to quantify LSC function in patients. In this proposal, we will establish these quantifiable in vitro parameters as in-process quality controls for the production of cLSCs and establish in vivo parameters as clinical outcome measures to objectively assess clinical outcomes. A pilot clinical study to evaluate the safety and efficacy of cLSCs produced under GMP condition will be conducted. Implementation of these in vitro and in vivo quantifiable parameters will make it possible to establish a standardized patient- specific stem cell therapy. The knowledge learned from the proposed study will form the foundation for the development of the next generation patient-specific limbal stem cell therapy.