Department of Ophthalmology, The Key Laboratory of Advanced Interdisciplinary Studies Center, The First Affiliated Hospital of Guangzhou Medical University, Guangzhou, Guangdong, China.
State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-Sen University, Guangzhou, Guangdong, China.
Invest Ophthalmol Vis Sci. 2024 Aug 1;65(10):12. doi: 10.1167/iovs.65.10.12.
The role of specific extracellular matrix (ECM) molecules in lens cell development and regeneration is poorly understood, as appropriate cellular models are lacking. Here, a laminin-based lens cell in vitro induction system was developed to study the role of laminin in human lens epithelial stem/progenitor cell (LES/PC) development.
The human embryonic stem cell-based lens induction system followed a three-stage protocol. The expression profile of laminins during lens induction was screened, and laminin-511 (LN511) was tested as a candidate substitute. LN511 induction system cellular and molecular features, including induction efficiency, transcription factor expression related to different lens development stages, ECM alterations, and Hippo/YAP signaling, were evaluated.
LAMA5, LAMB1, and LAMC1 were highly expressed around the time of LES/PC derivation. We chose LN511 (product of LAMA5, LAMB1, and LAMC1) and found that it considerably enhanced lens cell induction efficiency, compared to that in Matrigel-coated culture, as more and larger lentoid bodies were detected. Notably, LES/PC induction efficiency improved by promoting lens specification-related transcription factor expression and cell proliferation. Transcriptome analysis revealed that compared to those with Matrigel, ECM accumulation and cell adhesion were downregulated in the LN511 system. Hippo/YAP signaling was hypoactive during LES/P-like cell generation, and small molecule inhibitors of YAP/TAZ activity upregulated LES/PC marker expression and promoted the efficiency of LES/P-like cell derivation.
The laminin isoform LN511 is a reliable substitute for the LES/P-like cell induction system, and LN511-YAP acted as efficient modulators of LES/PC derivation; this contributes to knowledge of the role of the ECM in human lens development.
特定细胞外基质（ECM）分子在晶状体细胞发育和再生中的作用尚不清楚，因为缺乏适当的细胞模型。在这里，开发了一种基于层粘连蛋白的体外晶状体细胞诱导系统，以研究层粘连蛋白在人晶状体上皮干细胞/祖细胞（LES/PC）发育中的作用。
基于人胚胎干细胞的晶状体诱导系统遵循三阶段方案。筛选了层粘连蛋白在晶状体诱导过程中的表达谱，并测试了层粘连蛋白-511（LN511）作为候选替代物。评估了 LN511 诱导系统的细胞和分子特征，包括诱导效率、与不同晶状体发育阶段相关的转录因子表达、ECM 改变和 Hippo/YAP 信号。
LAMA5、LAMB1 和 LAMC1 在 LES/PC 衍生时高度表达。我们选择了 LN511（LAMA5、LAMB1 和 LAMC1 的产物），并发现与 Matrigel 包被培养相比，它大大提高了晶状体细胞诱导效率，因为检测到更多和更大的晶状体小体。值得注意的是，通过促进与晶状体特化相关的转录因子表达和细胞增殖，提高了 LES/PC 诱导效率。转录组分析显示，与 Matrigel 相比，LN511 系统中 ECM 积累和细胞黏附下调。在 LES/P 样细胞生成过程中，Hippo/YAP 信号活性降低，YAP/TAZ 活性的小分子抑制剂上调 LES/PC 标志物表达并促进 LES/P 样细胞衍生的效率。
层粘连蛋白同工型 LN511 是 LES/P 样细胞诱导系统的可靠替代物，LN511-YAP 是 LES/PC 衍生的有效调节剂；这有助于了解 ECM 在人晶状体发育中的作用。
