Trichostatin A-Mediated Epigenetic Modulation Predominantly Triggers Transcriptomic Alterations in the Ex Vivo Expanded Equine Chondrocytes.
Abstract: Epigenetic mechanisms of gene regulation are important for the proper differentiation of cells used for therapeutic and regenerative purposes. The primary goal of the present study was to investigate the impacts of 5-aza-2' deoxycytidine (5-AZA-dc)- and/or trichostatin A (TSA)-mediated approaches applied to epigenomically modulate the ex vivo expanded equine chondrocytes maintained in monolayer culture on the status of chondrogenic cytodifferentiation at the transcriptome level. The results of next-generation sequencing of 3' mRNA-seq libraries on stimulated and unstimulated chondrocytes of the third passage showed no significant influence of 5-AZA-dc treatment. Chondrocytes stimulated with TSA or with a combination of 5-AZA-dc+TSA revealed significant expressional decline, mainly for genes encoding histone and DNA methyltransferases, but also for other genes, many of which are enriched in canonical pathways that are important for chondrocyte biology. The TSA- or 5-AZA-dc+TSA-induced upregulation of expanded chondrocytes included genes that are involved in histone hyperacetylation and also genes relevant to rheumatoid arthritis and inflammation. Chondrocyte stimulation experiments including a TSA modifier also led to the unexpected expression incrementation of genes encoding HDAC3, SIRT2, and SIRT5 histone deacetylases and the MBD1 CpG-binding domain protein, pointing to another function of the TSA agent besides its epigenetic-like properties. Based on the transcriptomic data, TSA stimulation seems to be undesirable for chondrogenic differentiation of passaged cartilaginous cells in a monolayer culture. Nonetheless, obtained transcriptomic results of TSA-dependent epigenomic modification of the ex vivo expanded equine chondrocytes provide a new source of data important for the potential application of epigenetically altered cells for transplantation purposes in tissue engineering of the equine skeletal system.
Publication Date: 2022-10-29 PubMed ID: 36361948PubMed Central: PMC9655705DOI: 10.3390/ijms232113168Google Scholar: Lookup
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Summary
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This study explores the effects of two epigenetic modulators, 5-aza-2′ deoxycytidine and trichostatin A, on horse cartilage cells grown in a lab. It found that trichostatin A impacted gene expression significantly, which could influence how these cells are used in regenerative therapies for the horse skeletal system.
Objective of the Research
- This research investigates the impact of using 5-AZA-dc and TSA to modify gene regulation in horse cartilage cells grown outside of an organism in a lab. The goal is to better understand how these alterations might impact the differentiation of cells into cartilage, an important process for regenerative treatments.
Research Approach and Methods Utilized
- Researchers treated the cartilage cells to be studied with 5-AZA-dc, TSA, or both and then analyzed them using next-generation sequencing of 3′ mRNA-seq libraries. This technique allows for deep investigation of gene expression in the treated and untreated (control) cells.
Key Findings from the Research
- There was no notable effect on gene expression from the treatment of 5-AZA-dc alone. However, cells exposed to TSA alone or a combination of 5-AZA-dc and TSA showed significant changes in gene expression. These changes occurred primarily in genes related to DNA and histone methylation, processes that are involved in gene regulation and expression. These genes play crucial roles in maintaining the biology of cartilage cells.
- The upregulated genes in treated cells were involved in histone hyperacetylation, another molecular process involved in gene regulation, and genes related to rheumatoid arthritis and inflammation.
- Treatment also unexpectedly increased the expression of genes notorious for being histone deacetylases and the MBD1 CpG-binding domain protein, implying that TSA could be possessing some additional functionalities apart from its epigenetic-like properties.
- The data suggests that TSA stimulation might not be beneficial for the differentiation of cells into cartilage in a single layer culture, a conclusion which may affect future efforts to develop therapeutic cell treatments.
Significance and Implications of the Study
- Despite some concerning findings about the use of TSA, the research contributes valuable knowledge about the potential use of epigenetically modified cells in regenerating damaged horse skeletal tissue. Such information can be instrumental for designing successful therapeutic strategies in the field of equine tissue engineering.
Cite This Article
APA
Ząbek T, Witarski W, Szmatoła T, Sawicki S, Mrozowicz J, Samiec M.
(2022).
Trichostatin A-Mediated Epigenetic Modulation Predominantly Triggers Transcriptomic Alterations in the Ex Vivo Expanded Equine Chondrocytes.
Int J Mol Sci, 23(21), 13168.
https://doi.org/10.3390/ijms232113168 Publication
Researcher Affiliations
- Department of Animal Molecular Biology, National Research Institute of Animal Production, Krakowska 1 Street, 32-083 Balice, Poland.
- Department of Animal Molecular Biology, National Research Institute of Animal Production, Krakowska 1 Street, 32-083 Balice, Poland.
- Department of Animal Molecular Biology, National Research Institute of Animal Production, Krakowska 1 Street, 32-083 Balice, Poland.
- University Centre of Veterinary Medicine, University of Agriculture in Kraków, Mickiewicza 24/28, 30-059 Kraków, Poland.
- University Centre of Veterinary Medicine, University of Agriculture in Kraków, Mickiewicza 24/28, 30-059 Kraków, Poland.
- Department of Animal Molecular Biology, National Research Institute of Animal Production, Krakowska 1 Street, 32-083 Balice, Poland.
- Department of Reproductive Biotechnology and Cryoconservation, National Research Institute of Animal Production, Krakowska 1 Street, 32-083 Balice, Poland.
MeSH Terms
- Animals
- Azacitidine / pharmacology
- Chondrocytes / metabolism
- Decitabine / pharmacology
- DNA Methylation
- Epigenesis, Genetic
- Epigenomics
- Histone Deacetylase Inhibitors
- Histones / metabolism
- Horses / genetics
- Hydroxamic Acids / pharmacology
- Transcriptome
Conflict of Interest Statement
The authors declare no conflict of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript; or in the decision to publish the results.
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