Mitogen-Activated Protein Kinase Kinase Kinase 1 Overexpression Disrupts Development of the Ocular Surface Epithelium

Mitogen-Activated Protein Kinase Kinase Kinase 1 (MAP3K1) is a key signaling molecule essential for eyelid closure during embryogenesis. In mice, <i>Map3k1</i> knockout leads to a fully penetrant eye-open at birth (EOB) phenotype due to disrupted MAPK signaling, abnormal epithelial diffe...

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Main Authors: Maureen Mongan, Bo Xiao, Antonius Christianto, Yueh-Chiang Hu, Ying Xia
Format: Article
Language:English
Published: MDPI AG 2025-06-01
Series:Cells
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Online Access:https://www.mdpi.com/2073-4409/14/12/894
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author Maureen Mongan
Bo Xiao
Antonius Christianto
Yueh-Chiang Hu
Ying Xia
author_facet Maureen Mongan
Bo Xiao
Antonius Christianto
Yueh-Chiang Hu
Ying Xia
author_sort Maureen Mongan
collection DOAJ
description Mitogen-Activated Protein Kinase Kinase Kinase 1 (MAP3K1) is a key signaling molecule essential for eyelid closure during embryogenesis. In mice, <i>Map3k1</i> knockout leads to a fully penetrant eye-open at birth (EOB) phenotype due to disrupted MAPK signaling, abnormal epithelial differentiation, and morphogenesis. To further explore the roles of MAP3K1 in ocular development, we generated a Cre-inducible gain-of-function transgenic mouse, designated as <i>Map3k1<sup>TG</sup></i>, and crossed it with Lens epithelial (<i>Le)-Cre</i> mice to drive MAP3K1 overexpression in developing ocular surface epithelium (OSE). <i>Map3k1<sup>TG</sup></i>;<i>Le-Cre</i> embryos exhibited ocular defects including premature eyelid closure, lens degeneration, and corneal edema. While corneal epithelial differentiation remained intact, the lens epithelium degenerated with lens formation compromised. Eyelid epithelium was markedly thickened, containing cells with aberrant keratin (K)14/K10 co-expression. Genetic rescue experiments revealed that <i>Map3k1<sup>TG</sup></i>;<i>Le-Cre</i> restored eyelid closure in <i>Map3k1</i> knockout mice, whereas MAP3K1 deficiency attenuated the epithelial thickening caused by transgene expression. Mechanistically, MAP3K1 overexpression enhanced c-Jun phosphorylation in vivo and activated JNK-c-Jun, WNT, TGFβ, and Notch signaling and promoted keratinocyte proliferation and migration in vitro. These findings highlight a dose-sensitive role for MAP3K1 in regulating epithelial proliferation, differentiation, and morphogenesis during eyelid development.
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spelling doaj-art-128b32c4c2074768a4ba8a77987b5eb42025-08-20T03:26:20ZengMDPI AGCells2073-44092025-06-01141289410.3390/cells14120894Mitogen-Activated Protein Kinase Kinase Kinase 1 Overexpression Disrupts Development of the Ocular Surface EpitheliumMaureen Mongan0Bo Xiao1Antonius Christianto2Yueh-Chiang Hu3Ying Xia4Department of Environmental and Public Health Sciences, College of Medicine, University of Cincinnati, Cincinnati, OH 45267, USADepartment of Environmental and Public Health Sciences, College of Medicine, University of Cincinnati, Cincinnati, OH 45267, USADepartment of Environmental and Public Health Sciences, College of Medicine, University of Cincinnati, Cincinnati, OH 45267, USADivision of Developmental Biology, Cincinnati Children’s Hospital Medical Center, Cincinnati, OH 45267, USADepartment of Environmental and Public Health Sciences, College of Medicine, University of Cincinnati, Cincinnati, OH 45267, USAMitogen-Activated Protein Kinase Kinase Kinase 1 (MAP3K1) is a key signaling molecule essential for eyelid closure during embryogenesis. In mice, <i>Map3k1</i> knockout leads to a fully penetrant eye-open at birth (EOB) phenotype due to disrupted MAPK signaling, abnormal epithelial differentiation, and morphogenesis. To further explore the roles of MAP3K1 in ocular development, we generated a Cre-inducible gain-of-function transgenic mouse, designated as <i>Map3k1<sup>TG</sup></i>, and crossed it with Lens epithelial (<i>Le)-Cre</i> mice to drive MAP3K1 overexpression in developing ocular surface epithelium (OSE). <i>Map3k1<sup>TG</sup></i>;<i>Le-Cre</i> embryos exhibited ocular defects including premature eyelid closure, lens degeneration, and corneal edema. While corneal epithelial differentiation remained intact, the lens epithelium degenerated with lens formation compromised. Eyelid epithelium was markedly thickened, containing cells with aberrant keratin (K)14/K10 co-expression. Genetic rescue experiments revealed that <i>Map3k1<sup>TG</sup></i>;<i>Le-Cre</i> restored eyelid closure in <i>Map3k1</i> knockout mice, whereas MAP3K1 deficiency attenuated the epithelial thickening caused by transgene expression. Mechanistically, MAP3K1 overexpression enhanced c-Jun phosphorylation in vivo and activated JNK-c-Jun, WNT, TGFβ, and Notch signaling and promoted keratinocyte proliferation and migration in vitro. These findings highlight a dose-sensitive role for MAP3K1 in regulating epithelial proliferation, differentiation, and morphogenesis during eyelid development.https://www.mdpi.com/2073-4409/14/12/894MAP3K1 gain-of-functionocular surface epitheliumeyelid closureepithelial morphogenesis
spellingShingle Maureen Mongan
Bo Xiao
Antonius Christianto
Yueh-Chiang Hu
Ying Xia
Mitogen-Activated Protein Kinase Kinase Kinase 1 Overexpression Disrupts Development of the Ocular Surface Epithelium
Cells
MAP3K1 gain-of-function
ocular surface epithelium
eyelid closure
epithelial morphogenesis
title Mitogen-Activated Protein Kinase Kinase Kinase 1 Overexpression Disrupts Development of the Ocular Surface Epithelium
title_full Mitogen-Activated Protein Kinase Kinase Kinase 1 Overexpression Disrupts Development of the Ocular Surface Epithelium
title_fullStr Mitogen-Activated Protein Kinase Kinase Kinase 1 Overexpression Disrupts Development of the Ocular Surface Epithelium
title_full_unstemmed Mitogen-Activated Protein Kinase Kinase Kinase 1 Overexpression Disrupts Development of the Ocular Surface Epithelium
title_short Mitogen-Activated Protein Kinase Kinase Kinase 1 Overexpression Disrupts Development of the Ocular Surface Epithelium
title_sort mitogen activated protein kinase kinase kinase 1 overexpression disrupts development of the ocular surface epithelium
topic MAP3K1 gain-of-function
ocular surface epithelium
eyelid closure
epithelial morphogenesis
url https://www.mdpi.com/2073-4409/14/12/894
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AT antoniuschristianto mitogenactivatedproteinkinasekinasekinase1overexpressiondisruptsdevelopmentoftheocularsurfaceepithelium
AT yuehchianghu mitogenactivatedproteinkinasekinasekinase1overexpressiondisruptsdevelopmentoftheocularsurfaceepithelium
AT yingxia mitogenactivatedproteinkinasekinasekinase1overexpressiondisruptsdevelopmentoftheocularsurfaceepithelium