Histone chip-Seq identifies differential enhancer usage during chondrogenesis as critical for defining cell-type specificity
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Abstract
Epigenetic mechanisms are known to regulate gene expression during chondrogenesis. In this study, we have characterized the epigenome during the in vitro differentiation of human mesenchymal stem cells (hMSCs) into chondrocytes. Chromatin immunoprecipitation followed by next‐generation sequencing (ChIP‐seq) was used to assess a range of N‐terminal posttranscriptional modifications (marks) to histone H3 lysines (H3K4me3, H3K4me1, H3K27ac, H3K27me3, and H3K36me3) in both hMSCs and differentiated chondrocytes. Chromatin states were characterized using histone ChIP‐seq and cis ‐regulatory elements were identified in chondrocytes. Chondrocyte enhancers were associated with chondrogenesis‐related gene ontology (GO) terms. In silico analysis and integration of DNA methylation data with chondrogenesis chromatin states revealed that enhancers marked by histone marks H3K4me1 and H3K27ac were de‐methylated during in vitro chondrogenesis. Similarity analysis between hMSC and chondrocyte chromatin states defined in this study with epigenomes of cell‐types defined by the Roadmap Epigenomics project revealed that enhancers are more distinct between cell‐types compared to other chromatin states. Motif analysis revealed that the transcription factor SOX9 is enriched in chondrocyte enhancers. Luciferase reporter assays confirmed that chondrocyte enhancers characterized in this study exhibited enhancer activity which may be modulated by DNA methylation and SOX9 overexpression. Altogether, these integrated data illustrate the cross‐talk between different epigenetic mechanisms during chondrocyte differentiation.
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In chondrogenic systems, SOX9 can activate enhancers but may not be required for initiation of chromatin changes.
“In chondrogenic systems, SOX9 can activate enhancers (19), but may not be required for the initiation of chromatin changes (20).”
From SOX9 reprograms endothelial cells by altering the chromatin landscape (2022) · cites this paper as [19]
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- Authors
- Cheung, K., Barter, M. J., Falk, J., Proctor, C. J., Reynard, L. N., & Young, D. A.
- Affiliations
- Newcastle University (GB)
- Skeletal Research Group Institute of Genetic Medicine Newcastle University Central Parkway Newcastle upon Tyne UK
- Corresponding author
- Cheung, Kathleen
- Young, David A.
- Venue
- The FASEB Journal
- Publisher
- Wiley
- Published
- 2020-02-14
- Volume
- 34
- Issue
- 4
- Pages
- 5317-5331
- ISSN
- 0892-6638 (ISSN-L)
- 1530-6860
- PMC
- PMC7187454
- OpenAlex
- W3006007721
- Cited by
- 23
- References
- 104
- Citations by year
- 2026: 5 · 2025: 1 · 2024: 4 · 2023: 6 · 2022: 3 · 2021: 2 · 2020: 2
- FWCI
- 1.05
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- Locations
- The FASEB Journal (publishedVersion, cc-by)
- bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, other-oa)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- Newcastle University ePrints (Newcastle Univesity) (cc-by)
- Funders
- Medical Research Council
- Arthritis Research UK (18746, 19424)
- Centro de Investigación Médica Aplicada, Universidad de Navarra (JXR 10641, MR/P020941/1)
- JGW Patterson Foundation
- Dunhill Medical Trust
- National Institute for Health Research
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Genomics and Chromatin Dynamics
- Health Sciences › Medicine › Rheumatology › Osteoarthritis Treatment and Mechanisms
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Epigenetics and DNA Methylation
- Keywords
- chondrogenesis, chondrocytes, mesenchymal stem cells, histone modifications, enhancers, DNA methylation, H3K4me1, H3K27ac, SOX9, chromatin state, cis-regulatory elements, cell type specificity, epigenetic regulation
- MeSH terms
- Chromatin Immunoprecipitation Sequencing, Adult, Cell Differentiation, Cells, Cultured, Chromatin, Enhancer Elements, Genetic, Female, Histones, Humans, Promoter Regions, Genetic, Cell Lineage, DNA Methylation, Chondrocytes, Chondrogenesis, Epigenesis, Genetic, SOX9 Transcription Factor, Young Adult, Epigenomics
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