June 2022
Volume 63, Issue 7
Open Access
ARVO Annual Meeting Abstract  |   June 2022
The cAMP Response Element Binding Protein (CREB) Negatively Regulates Pax6 Expression in Lens Epithelial Cells through Its Direct Binding to the Core Promoter of Pax6 Gene
Author Affiliations & Notes
  • Ling Wang
    Changsha Medical University, Changsha, Hunan, China
    Sun Yat-Sen University Zhongshan Ophthalmic Center State Key Laboratory of Ophthalmology, Guangzhou, Guangdong, China
  • Jialing Fu
    Sun Yat-Sen University Zhongshan Ophthalmic Center State Key Laboratory of Ophthalmology, Guangzhou, Guangdong, China
  • Yan Wang
    Sun Yat-Sen University Zhongshan Ophthalmic Center State Key Laboratory of Ophthalmology, Guangzhou, Guangdong, China
  • Shuyu Zheng
    Sun Yat-Sen University Zhongshan Ophthalmic Center State Key Laboratory of Ophthalmology, Guangzhou, Guangdong, China
  • yuan xiao
    Sun Yat-Sen University Zhongshan Ophthalmic Center State Key Laboratory of Ophthalmology, Guangzhou, Guangdong, China
  • Jingmiao Wang
    Sun Yat-Sen University Zhongshan Ophthalmic Center State Key Laboratory of Ophthalmology, Guangzhou, Guangdong, China
  • Jiawen Xiang
    Sun Yat-Sen University Zhongshan Ophthalmic Center State Key Laboratory of Ophthalmology, Guangzhou, Guangdong, China
  • David W Li
    Sun Yat-Sen University Zhongshan Ophthalmic Center State Key Laboratory of Ophthalmology, Guangzhou, Guangdong, China
  • Footnotes
    Commercial Relationships   Ling Wang None; Jialing Fu None; Yan Wang None; Shuyu Zheng None; yuan xiao None; Jingmiao Wang None; Jiawen Xiang None; David Li None
  • Footnotes
    Support  Supported by National Natural Science Foundation of China (Grants #82000842, #81970787, #82000876, #81770910) and Natural Science Foundation of Guangdong Province and Guangdong City Joint Program of China (2019B1515120014),and the Fundamental Funds, 3030901010110 of the State Key Laboratory of Ophthalmology of Zhongshan Ophthalmic Center.
Investigative Ophthalmology & Visual Science June 2022, Vol.63, 661 – F0016. doi:
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    • Get Citation

      Ling Wang, Jialing Fu, Yan Wang, Shuyu Zheng, yuan xiao, Jingmiao Wang, Jiawen Xiang, David W Li; The cAMP Response Element Binding Protein (CREB) Negatively Regulates Pax6 Expression in Lens Epithelial Cells through Its Direct Binding to the Core Promoter of Pax6 Gene. Invest. Ophthalmol. Vis. Sci. 2022;63(7):661 – F0016.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose : CREB is a transcription factor and its most prominent function is to control the synaptic plasticity associated with long-term memory. In our recent studies, we demonstrated that CREB regulates oxidative stress-induced apoptosis and aging in lens by inhibiting αB-crystallin expression but promoting P300-P53-Bak/Bax signaling axis (Wang L et al. 2021. Aging Cell). Whether CREB is capable to regulate lens differentiation remains largely unknown. Here, in the present study, we demonstrate that CREB directly regulates Pax6 and other genes to control differentiation of mouse lens.

Methods : CRISPR/Cas9 Technology was used to create mouse model CREB-S133A. RNAseq analysis was used to compare the transcriptional activity of wild type and mutant CREB-S133A.Gel mobility shifting assays were used to determine binding of CREB to Pax6 gene promoter. ChIP assays were used to confirm the binding of CREB to the Pax6 gene promoter. Reporter gene constructs driven by different promoter and enhancer regions of the Pax6 gene were generated and used to assay the relative strength of different Pax6 gene promoter regions. QRT-PCR and Wes were used to analyze mRNA and protein expression levels.

Results : RNAseq analysis revealed the reverse relationship between CREB and Pax6. EMSA assays demonstrate that CREB directly binds to the promoter region of Pax6. ChIP assay results confirmed that CREB binds to Pax6 promoter in vivo. In mouse lens epithelial cell, overexpression WT-CREB resulted in downregulation of Pax6. Silence of CREB upregulates Pax6. RNAseq analysis also revealed that CREB can regulate a panel of differentiation-related genes.

Conclusions : Through direct control of Pax6 and other downstream differentiation-related genes, CREB regulates lens differentiation. Supported by National Natural Science Foundation of China (Grants #82000842, #81970787, #82000876, #81770910) and Natural Science Foundation of Guangdong Province and Guangdong City Joint Program of China (2019B1515120014),and the Fundamental Funds, 3030901010110 of the State Key Laboratory of Ophthalmology of Zhongshan Ophthalmic Center.

This abstract was presented at the 2022 ARVO Annual Meeting, held in Denver, CO, May 1-4, 2022, and virtually.

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