Acta Biomaterialia ( IF 9.4 ) Pub Date : 2018-05-17 , DOI: 10.1016/j.actbio.2018.05.023 Majed Felemban 1 , Birthe Dorgau 1 , Nicola Claire Hunt 1 , Dean Hallam 1 , Darin Zerti 1 , Roman Bauer 2 , Yuchun Ding 3 , Joseph Collin 1 , David Steel 1 , Natalio Krasnogor 3 , Jumana Al-Aama 4 , Susan Lindsay 1 , Carla Mellough 5 , Majlinda Lako 1
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The extracellular matrix (ECM) plays an important role in numerous processes including cellular proliferation, differentiation, migration, maturation, adhesion guidance and axonal growth. To date, there has been no detailed analysis of the ECM distribution during retinal ontogenesis in humans and the functional importance of many ECM components is poorly understood. In this study, the expression of key ECM components in adult mouse and monkey retina, developing and adult human retina and retinal organoids derived from human pluripotent stem cells was studied. Our data indicate that basement membrane ECMs (Fibronectin and Collagen IV) were expressed in Bruch’s membrane and the inner limiting membrane of the developing human retina, whilst the hyalectins (Versican and Brevican), cluster of differentiation 44 (CD44), photoreceptor-specific ECMs Interphotoreceptor Matrix Proteoglycan 1 (IMPG1) and Interphotoreceptor Matrix Proteoglycan 2 (IMPG2) were detected in the developing interphotoreceptor matrix (IPM). The expression of IMPG1, Versican and Brevican in the developing IPM was conserved between human developing retina and human pluripotent stem cell-derived retinal organoids. Blocking the action of CD44 and IMPG1 in pluripotent stem cell derived retinal organoids affected the development of photoreceptors, their inner/outer segments and connecting cilia and disrupted IPM formation, with IMPG1 having an earlier and more significant impact. Together, our data suggest an important role for IMPG1 and CD44 in the development of photoreceptors and IPM formation during human retinogenesis.
Statement of Significance
The expression and the role of many extracellular matrix (ECM) components during human retinal development is not fully understood. In this study, expression of key ECM components (Collagen IV, Fibronectin, Brevican, Versican, IMPG1 and IMPG2) was investigated during human retinal ontogenesis. Collagen IV and Fibronectin were expressed in Bruch’s membrane; whereas Brevican, Versican, IMPG1 & IMPG2 in the developing interphotoreceptor matrix (IPM). Retinal organoids were successfully generated from pluripotent stem cells. The expression of ECM components was examined in the retinal organoids found to recapitulate human retinal development in vivo. Using functional blocking experiments, this study also highlight an important role of IMPG1 and CD44 in the development of photoreceptors and IPM formation.
中文翻译：
人多能干细胞衍生的视网膜类器官中的细胞外基质表达概括了体内的视网膜发生，并揭示了 IMPG1 和 CD44 在光感受器和光感受器间基质发育中的重要作用
细胞外基质 (ECM) 在许多过程中发挥重要作用，包括细胞增殖、分化、迁移、成熟、粘附引导和轴突生长。迄今为止，还没有对人类视网膜个体发育过程中 ECM 分布的详细分析，并且对许多 ECM 成分的功能重要性知之甚少。在这项研究中，研究了成年小鼠和猴子视网膜、发育中和成年人类视网膜以及源自人类多能干细胞的视网膜类器官中关键 ECM 成分的表达。我们的数据表明基底膜 ECM（纤连蛋白和胶原蛋白 IV）在布鲁赫膜和发育中的人类视网膜的内界膜中表达，而透明凝集素（Versican 和 Brevican）、分化簇 44（CD44）、在显影的光感受器间基质 (IPM) 中检测到光感受器特异性 ECMs Interphotoreceptor Matrix Proteoglycan 1 (IMPG1) 和 Interphotoreceptor Matrix Proteoglycan 2 (IMPG2)。IMPG1、Versican 和 Brevican 在发育中的 IPM 中的表达在人类发育中的视网膜和人类多能干细胞衍生的视网膜类器官之间是保守的。在多能干细胞衍生的视网膜类器官中阻断 CD44 和 IMPG1 的作用会影响光感受器、它们的内/外段和连接纤毛的发育，并破坏 IPM 的形成，其中 IMPG1 具有更早和更显着的影响。总之，我们的数据表明 IMPG1 和 CD44 在人类视网膜发生过程中光感受器发育和 IPM 形成中的重要作用。
重要性声明
许多细胞外基质 (ECM) 成分在人类视网膜发育过程中的表达和作用尚不完全清楚。在这项研究中，研究了人类视网膜个体发育过程中关键 ECM 成分（胶原蛋白 IV、纤连蛋白、Brevican、Versican、IMPG1 和 IMPG2）的表达。胶原蛋白 IV 和纤连蛋白在布鲁赫膜中表达；而 Brevican、Versican、IMPG1 和 IMPG2 在显影间感光器基质 (IPM) 中。从多能干细胞成功地产生了视网膜类器官。ECM 成分的表达在视网膜类器官中进行了检查，发现这些类器官可以概括人体视网膜的体内发育。通过功能性阻断实验，本研究还强调了 IMPG1 和 CD44 在感光细胞发育和 IPM 形成中的重要作用。
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