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Yap1 is Essential for Lens Development and Exhibits Dose-Dependent Effects

ABSTRACT: Purpose: The Hippo-YAP pathway, which is crucial for regulating organ size, has been shown to play critical roles in lens development and homeostasis. However, Yap1's specific role in early lens morphogenesis is unclear. This study aims to investigate lens defects in Yap1 conditional knockout mice and explore the potential mechanisms. Methods: Lens-specific Yap1 deletion was induced using P0-3.9GFPCre in both Yap1fl/fl and Yap1fl/+ mice. Slit-lamp and brightfield imaging were used to observe the ocular phenotype. Lens histology was examined by H&E staining, while marker expression was analyzed using immunofluorescence and Western blot. Cellular proliferation and apoptosis were assessed via BrdU and TUNEL assays, respectively. RNA-seq was performed to profile the lens transcriptome. Results: Yap1 was consistently expressed throughout lens development. In contrast to previous reports, Yap1 deletion produced a continuous spectrum of lens defects, ranging from normal lenses to microphakia and, in the most severe cases, aphakia. The severity of these abnormalities exhibited a clear dose-dependent relationship with residual YAP1 protein levels. Notably, we identified lens–cornea adhesion as a previously unreported phenotype in Yap1-deficient mice, which may associate with reduced expression of the transcription factors FoxE3 and AP2α. The reduction in lens size was primarily attributable to impaired proliferation and a modest increase in apoptosis in lens epithelial cells. Conclusion: Our study identifies Yap1 as an essential regulator of early lens morphogenesis, acting through a finely tuned and highly dosage-sensitive regulatory mechanism.

ORGANISM(S): Mus musculus

PROVIDER: GSE315383 | GEO | 2026/05/20

REPOSITORIES: GEO

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Project description:FOXE3 is a lens specific transcription factor that has been associated with anterior segment ocular dysgenesis. To determine the transcriptional target(s) of FOXE3 that are indispensable for the anterior segment development, we examined the transcriptome and the proteome of cells expressing truncated FOXE3 responsible for Peters anomaly identified through linkage-coupled next-generation whole exome sequencing. We found that DNAJB1, an autophagy-associated protein, was the only candidate exhibiting differential expression in both screens. We confirmed the candidacy of DNAJB1 through chromatin immunoprecipitation and luciferase assays while knockdown of DNAJB1 in human lens epithelial cells resulted in mitotic arrest. Subsequently, we targeted dnajb1a in zebrafish through injection of a splice-blocking morpholino. The dnajb1a morphants exhibited underdeveloped cataractous lenses with persistent apoptotic nuclei. In conclusion, we have identified DNAJB1 as a transcriptional target of FOXE3 in a novel pathway that is crucial for development of the anterior segment of the eye. Human Embryonic Kidney (HEK293FT) cells were transfected with the expression vector (pT-RexTM-DEST30) harboring either the wild type or the mutant (C240*) FOXE3 ORF (open reading frame). The experimental design included a total of eight biological replicates of cells expressing the wild type and eight replicates of mutant FOXE3 along with eight non-transfected controls. Cells were harvested 24-hour post-transfection and subjected to total RNA isolation for the preparation of whole transcriptome next-generation sequencing libraries. Initially, we examined the quality of transcriptome libraries on a MiSeq genome analyzer. Subsequent to confirmation of the quality, all libraries were paired-end sequenced (2 x 100 bp) using Illumina TruSeq Cluster V3 flow cell at a concentration of 13.0 pM in two separate lanes (12 bar-coded mRNA pooled libraries in each lane) on a HiSeq 2000 genome analyzer.

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Yap1 is Essential for Lens Development and Exhibits Dose-Dependent Effects

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