Aberrant TGF-β1 signaling activation by MAF underlies pathological lens growth in high myopia

High myopia is a leading cause of blindness worldwide. Myopia progression may lead to pathological changes of lens and affect the outcome of lens surgery, but the underlying mechanism remains unclear. Here, we find an increased lens size in highly myopic eyes associated with up-regulation of β/γ-crystallin expressions. Similar findings are replicated in two independent mouse models of high myopia. Mechanistic studies show that the transcription factor MAF plays an essential role in up-regulating β/γ-crystallins in high myopia, by direct activation of the crystallin gene promoters and by activation of TGF-β1-Smad signaling. Our results establish lens morphological and molecular changes as a characteristic feature of high myopia, and point to the dysregulation of the MAF-TGF-β1-crystallin axis as an underlying mechanism, providing an insight for therapeutic interventions.


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12/27/2020
For MRI data collection, sample size was calculated based on results of preliminary investigation using G*Power Software 3.1. Lens volume data were collected using an open-source software (3D Slicer 4.10.1 r27931, https://www.slicer.org/). The axial length and the maximum cross-sectional area of lens of mice were measured with ParaVision ® 6.0.1 software (Bruker, USA).
Most statistical analyses were conducted using the Prism 8.0 (GraphPad Software, Inc., USA). For microarray, differential gene expression was determined using the limma statistical package 3.26.9 as described at http:// www.bioconductor.org.
Human microarray data have been deposited in the Gene Expression Omnibus (GEO) under the accession code GSE136701. Online Jaspar database (http://jaspar.genereg.net/) was used for prediction of MAF binding sites in promoters of crystallin genes. The source data underlying figures shown in the manuscript and the raw images for the immunoblots are provided as a Source Data file.

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October 2018 Field-specific reporting Please select the one below that is the best fit for your research. If you are not sure, read the appropriate sections before making your selection. No data were excluded.
At least three replications were made in all experiments. And all attempts at replication were successful.
No randomization of mice. For establishement of defocus-induced myopic mouse model, male C57BL/6J mice were subjected to continuous defocus all to the right eye. Wild-type mice and Irbp KO mice in comparison were age-and sex-matched. For experiments using specimens with certain treatment, our samples were consecutively collected and randomly picked when deciding which one to use in a specific assay or to form a pooled sample or to be treated.
For MRI data collection, researchers did not know the diagnosis of high myopia or emmetropia before measurement, however axial length and lens dimensions were measured simultaneously in the same image, thus blinding could not be done here. Note that full information on the approval of the study protocol must also be provided in the manuscript.