Abe-Fukasawa Natsuki, Hayashi Ryuhei, Morita Mio, Azuma Shohei, Iwawaki Takumi, Kagaya Kenta, Nishino Taito, Nishida Kohji
Biological Research Laboratories, Nissan Chemical Corporation, Shiraoka, Saitama, 349-0294, Japan.
Department of Ophthalmology, The University of Osaka Graduate School of Medicine, Suita, Osaka, 565-0871, Japan.
Regen Ther. 2025 Sep 3;30:730-739. doi: 10.1016/j.reth.2025.08.014. eCollection 2025 Dec.
Transplantation of expanded corneal endothelial cells (CECs) has been regarded as a promising approach for treating corneal diseases caused by CEC damage or dysfunction. However, an efficient method for expanding CECs remains inadequately established.
We examined whether small molecule inhibitors of large tumor suppressor kinase (LATS) promote the proliferation of CECs. We also evaluated the expression of functional markers in CECs treated with the inhibitors.
We found that LATS kinase inhibitors enhance the cell density of bovine CECs . CECs that were expanded in the presence of these inhibitors exhibited increased nuclear translocation of yes-associated protein (YAP) and upregulated expression of YAP-regulated genes. Furthermore, we observed that YAP was essential for promoting cell proliferation. Notably, the inhibitors also increased the density of primary human CECs. Expanded human CECs expressed CEC functional markers, including Na/K-transporting ATPase subunit alpha-1 (ATP1A1), Zonula occludens-1 (ZO-1), and N-cadherin; they showed upregulated expression of YAP-regulated genes.
Collectively, these findings support the development of efficient culture techniques for CEC expansion and may facilitate the advancement of therapeutic strategies for CEC-associated diseases.
扩增角膜内皮细胞（CECs）移植被视为治疗由CEC损伤或功能障碍引起的角膜疾病的一种有前景的方法。然而，一种有效的CECs扩增方法仍未充分建立。
我们研究了大肿瘤抑制激酶（LATS）的小分子抑制剂是否能促进CECs的增殖。我们还评估了用这些抑制剂处理的CECs中功能标志物的表达。
我们发现LATS激酶抑制剂可提高牛CECs的细胞密度。在这些抑制剂存在下扩增的CECs表现出Yes相关蛋白（YAP）核转位增加以及YAP调控基因的表达上调。此外，我们观察到YAP对促进细胞增殖至关重要。值得注意的是，这些抑制剂还增加了原代人CECs的密度。扩增的人CECs表达CEC功能标志物，包括钠/钾转运ATP酶α-1亚基（ATP1A1）、紧密连接蛋白1（ZO-1）和N-钙黏蛋白；它们显示出YAP调控基因的表达上调。
总的来说，这些发现支持开发用于CECs扩增的高效培养技术，并可能促进CEC相关疾病治疗策略的进展。
