Lineage Cell Therapeutics, Inc., Alameda, California, USA.
Department of Small Animal Clinical Sciences, College of Veterinary Medicine, Michigan State University, East Lansing, Michigan, USA.
Stem Cells Dev. 2021 Apr;30(8):399-417. doi: 10.1089/scd.2020.0085.
Progressive vision loss, caused by retinal degenerative (RD) diseases such as age-related macular degeneration, retinitis pigmentosa, and Leber congenital amaurosis, severely impacts quality of life and affects millions of people. Finding efficient treatment for blinding diseases is among the greatest unmet clinical needs. The evagination of optic vesicles from developing pluripotent stem cell-derived neuroepithelium and self-organization, lamination, and differentiation of retinal tissue in a dish generated considerable optimism for developing innovative approaches for treating RD diseases, which previously were not feasible. Retinal organoids may be a limitless source of multipotential retinal progenitors, photoreceptors (PRs), and the whole retinal tissue, which are productive approaches for developing RD disease therapies. In this study we compared the distribution and expression level of molecular markers (genetic and epigenetic) in human fetal retina (age 8-16 weeks) and human embryonic stem cell (hESC)-derived retinal tissue (organoids) by immunohistochemistry, RNA-seq, flow cytometry, and mass-spectrometry (to measure methylated and hydroxymethylated cytosine level), with a focus on PRs to evaluate the clinical application of hESC-retinal tissue for vision restoration. Our results revealed high correlation in gene expression profiles and histological profiles between human fetal retina (age 8-13 weeks) and hESC-derived retinal tissue (10-12 weeks). The transcriptome signature of hESC-derived retinal tissue from retinal organoids maintained for 24 weeks in culture resembled the transcriptome of human fetal retina of more advanced developmental stages. The histological profiles of 24 week-old hESC-derived retinal tissue displayed mature PR immunophenotypes and presence of developing inner and outer segments. Collectively, our work highlights the similarity of hESC-derived retinal tissue at early stages of development (10 weeks), and human fetal retina (age 8-13 weeks) and it supports the development of regenerative medicine therapies aimed at using tissue from hESC-derived retinal organoids (hESC-retinal implants) for mitigating vision loss.
进行性视力丧失是由视网膜退行性疾病引起的，例如年龄相关性黄斑变性、色素性视网膜炎和莱伯先天性黑矇，严重影响生活质量，影响了数百万人。寻找有效的致盲疾病治疗方法是最大的未满足的临床需求之一。从多能干细胞衍生的神经外胚层中诱导出的视泡外凸以及视网膜组织在培养皿中的自我组织、分层和分化，为治疗视网膜退行性疾病带来了新的希望，因为这些疾病以前是无法治疗的。视网膜类器官可能是多能视网膜祖细胞、光感受器 (PR) 和整个视网膜组织的无限来源，这为开发治疗视网膜退行性疾病的方法提供了有希望的途径。在这项研究中，我们通过免疫组织化学、RNA-seq、流式细胞术和质谱（测量甲基化和羟甲基化胞嘧啶水平）比较了人胎龄视网膜（8-16 周）和人胚胎干细胞（hESC）衍生的视网膜组织（类器官）中分子标志物（遗传和表观遗传）的分布和表达水平，重点是 PR，以评估 hESC 视网膜组织在视力恢复中的临床应用。我们的结果表明，人胎龄视网膜（8-13 周）和 hESC 衍生的视网膜组织（10-12 周）之间的基因表达谱和组织学特征高度相关。在培养中培养 24 周的视网膜类器官衍生的 hESC 衍生的视网膜组织的转录组特征与更成熟的发育阶段的人胎龄视网膜的转录组特征相似。24 周龄的 hESC 衍生的视网膜组织的组织学特征显示出成熟的 PR 免疫表型和正在发育的内节和外节的存在。总的来说，我们的工作强调了 hESC 衍生的视网膜组织在早期发育阶段（10 周）与人类胎龄视网膜（8-13 周）的相似性，支持了旨在使用 hESC 衍生的视网膜类器官（hESC-视网膜植入物）组织开发再生医学疗法的发展，以减轻视力丧失。
