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Integrated Transcriptome and Proteome Analyses Reveal the Regulatory Role of miR-146a in Human Limbal Epithelium via Notch Signaling
MiR-146a is upregulated in the stem cell-enriched limbal region vs. central human cornea and can mediate corneal epithelial wound healing. The aim of this study was to identify miR-146a targets in human primary limbal epithelial cells (LECs) using genomic and proteomic analyses. RNA-seq combined wit...
Autores principales: | , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7650592/ https://www.ncbi.nlm.nih.gov/pubmed/32993109 http://dx.doi.org/10.3390/cells9102175 |
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author | Poe, Adam J. Kulkarni, Mangesh Leszczynska, Aleksandra Tang, Jie Shah, Ruchi Jami-Alahmadi, Yasaman Wang, Jason Kramerov, Andrei A. Wohlschlegel, James Punj, Vasu Ljubimov, Alexander V. Saghizadeh, Mehrnoosh |
author_facet | Poe, Adam J. Kulkarni, Mangesh Leszczynska, Aleksandra Tang, Jie Shah, Ruchi Jami-Alahmadi, Yasaman Wang, Jason Kramerov, Andrei A. Wohlschlegel, James Punj, Vasu Ljubimov, Alexander V. Saghizadeh, Mehrnoosh |
author_sort | Poe, Adam J. |
collection | PubMed |
description | MiR-146a is upregulated in the stem cell-enriched limbal region vs. central human cornea and can mediate corneal epithelial wound healing. The aim of this study was to identify miR-146a targets in human primary limbal epithelial cells (LECs) using genomic and proteomic analyses. RNA-seq combined with quantitative proteomics based on multiplexed isobaric tandem mass tag labeling was performed in LECs transfected with miR-146a mimic vs. mimic control. Western blot and immunostaining were used to confirm the expression of some targeted genes/proteins. A total of 251 differentially expressed mRNAs and 163 proteins were identified. We found that miR-146a regulates the expression of multiple genes in different pathways, such as the Notch system. In LECs and organ-cultured corneas, miR-146a increased Notch-1 expression possibly by downregulating its inhibitor Numb, but decreased Notch-2. Integrated transcriptome and proteome analyses revealed the regulatory role of miR-146a in several other processes, including anchoring junctions, TNF-α, Hedgehog signaling, adherens junctions, TGF-β, mTORC2, and epidermal growth factor receptor (EGFR) signaling, which mediate wound healing, inflammation, and stem cell maintenance and differentiation. Our results provide insights into the regulatory network of miR-146a and its role in fine-tuning of Notch-1 and Notch-2 expressions in limbal epithelium, which could be a balancing factor in stem cell maintenance and differentiation. |
format | Online Article Text |
id | pubmed-7650592 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-76505922020-11-10 Integrated Transcriptome and Proteome Analyses Reveal the Regulatory Role of miR-146a in Human Limbal Epithelium via Notch Signaling Poe, Adam J. Kulkarni, Mangesh Leszczynska, Aleksandra Tang, Jie Shah, Ruchi Jami-Alahmadi, Yasaman Wang, Jason Kramerov, Andrei A. Wohlschlegel, James Punj, Vasu Ljubimov, Alexander V. Saghizadeh, Mehrnoosh Cells Article MiR-146a is upregulated in the stem cell-enriched limbal region vs. central human cornea and can mediate corneal epithelial wound healing. The aim of this study was to identify miR-146a targets in human primary limbal epithelial cells (LECs) using genomic and proteomic analyses. RNA-seq combined with quantitative proteomics based on multiplexed isobaric tandem mass tag labeling was performed in LECs transfected with miR-146a mimic vs. mimic control. Western blot and immunostaining were used to confirm the expression of some targeted genes/proteins. A total of 251 differentially expressed mRNAs and 163 proteins were identified. We found that miR-146a regulates the expression of multiple genes in different pathways, such as the Notch system. In LECs and organ-cultured corneas, miR-146a increased Notch-1 expression possibly by downregulating its inhibitor Numb, but decreased Notch-2. Integrated transcriptome and proteome analyses revealed the regulatory role of miR-146a in several other processes, including anchoring junctions, TNF-α, Hedgehog signaling, adherens junctions, TGF-β, mTORC2, and epidermal growth factor receptor (EGFR) signaling, which mediate wound healing, inflammation, and stem cell maintenance and differentiation. Our results provide insights into the regulatory network of miR-146a and its role in fine-tuning of Notch-1 and Notch-2 expressions in limbal epithelium, which could be a balancing factor in stem cell maintenance and differentiation. MDPI 2020-09-26 /pmc/articles/PMC7650592/ /pubmed/32993109 http://dx.doi.org/10.3390/cells9102175 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Poe, Adam J. Kulkarni, Mangesh Leszczynska, Aleksandra Tang, Jie Shah, Ruchi Jami-Alahmadi, Yasaman Wang, Jason Kramerov, Andrei A. Wohlschlegel, James Punj, Vasu Ljubimov, Alexander V. Saghizadeh, Mehrnoosh Integrated Transcriptome and Proteome Analyses Reveal the Regulatory Role of miR-146a in Human Limbal Epithelium via Notch Signaling |
title | Integrated Transcriptome and Proteome Analyses Reveal the Regulatory Role of miR-146a in Human Limbal Epithelium via Notch Signaling |
title_full | Integrated Transcriptome and Proteome Analyses Reveal the Regulatory Role of miR-146a in Human Limbal Epithelium via Notch Signaling |
title_fullStr | Integrated Transcriptome and Proteome Analyses Reveal the Regulatory Role of miR-146a in Human Limbal Epithelium via Notch Signaling |
title_full_unstemmed | Integrated Transcriptome and Proteome Analyses Reveal the Regulatory Role of miR-146a in Human Limbal Epithelium via Notch Signaling |
title_short | Integrated Transcriptome and Proteome Analyses Reveal the Regulatory Role of miR-146a in Human Limbal Epithelium via Notch Signaling |
title_sort | integrated transcriptome and proteome analyses reveal the regulatory role of mir-146a in human limbal epithelium via notch signaling |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7650592/ https://www.ncbi.nlm.nih.gov/pubmed/32993109 http://dx.doi.org/10.3390/cells9102175 |
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