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Home / Equine Research Bank / Arthritis Rheumatol / Transcriptome-wide analysis of messenger RNA decay in normal...
Arthritis & rheumatology (Hoboken, N.J.)2014; 66(11); 3052-3061; doi: 10.1002/art.38849

Transcriptome-wide analysis of messenger RNA decay in normal and osteoarthritic human articular chondrocytes.

Abstract: Messenger RNA (mRNA) decay rates control not only gene expression levels, but also responsiveness to altered transcriptional input. We undertook this study to examine transcriptome-wide posttranscriptional regulation in both normal and osteoarthritic (OA) human articular chondrocytes. Methods: Human articular chondrocytes were isolated from normal or OA tissue. Equine articular chondrocytes were isolated from young or old horses at a commercial abattoir. RNA decay was measured across the transcriptome in human cells by microarray analysis following an actinomycin D chase. Messenger RNA levels in samples were confirmed using quantitative reverse transcription-polymerase chain reaction. Results: Examination of total mRNA expression levels demonstrated significant differences in the expression of transcripts between normal and OA chondrocytes. Interestingly, almost no difference was observed in total mRNA expression between chondrocytes from intact OA cartilage and those from fibrillated OA cartilage. Decay analysis revealed a set of rapidly turned over transcripts associated with transcriptional control and programmed cell death that were common to all chondrocytes and contained binding sites for abundant cartilage microRNAs. Many transcripts exhibited altered mRNA half-lives in human OA chondrocytes compared to normal cells. Specific transcripts whose decay rates were altered were generally less stable in these pathologic cells. Examination of selected genes in chondrocytes from young and old healthy horses did not identify any change in mRNA turnover. Conclusions: This is the first investigation into the "posttranscriptome" of the chondrocyte. It identifies a set of short-lived chondrocyte mRNAs likely to be highly responsive to altered transcriptional input as well as mRNAs whose decay rates are affected in OA chondrocytes.
© 2014 The Authors. Arthritis & Rheumatology is published by Wiley Periodicals, Inc. on behalf of the American College of Rheumatology.
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Publication Date: 2014-08-27 PubMed ID: 25155964PubMed Central: PMC4233952DOI: 10.1002/art.38849Google Scholar: Lookup
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  • Comparative Study
  • Journal Article
  • Research Support
  • Non-U.S. Gov't

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research paper essentially studies how messenger RNA (mRNA) decay rates vary and affect gene expression levels in both normal and osteoarthritic (OA) human articular chondrocytes, which are cells that are part of cartilage. This investigation of the ‘posttranscriptome’ reveals key differences in mRNA decay rates between normal cells and OA cells.

Methods

  • In the study, human articular chondrocytes – cells derived from the cartilage that lines the joints – were obtained from both normal and osteoarthritic tissues.
  • Equine (horse) articular chondrocytes were also collected from both young and old horses at a commercial abattoir, to compare the results.
  • A procedure called an actinomycin D chase was used for measuring the decay of RNA across the transcriptome in human cells.
  • Confirmation of mRNA levels in the samples was done using a method known as quantitative reverse transcription-polymerase chain reaction.

Results

  • The analysis of total mRNA expression levels showed significant differences in the expression of transcripts between normal and osteoarthritic (OA) chondrocytes.
  • It was noted, however, that there was minimal difference in total mRNA expression when OA chondrocytes were compared, whether they were from intact OA cartilage or fibrillated (fragmented) OA cartilage.
  • The decay analysis exposed that there was a group of quickly turned over transcripts connected to transcriptional control and programmed cell death that were common to all chondrocytes.
  • The study found alteration in the mRNA half-lives in human OA chondrocytes as compared to normal cells, indicating less stability of specific transcripts in pathologic cells.
  • When comparing decay rates in chondrocytes from young and old healthy horses, no change in mRNA turnover was observed.

Conclusions

  • This was a pioneering study into the ‘posttranscriptome’ of the chondrocyte, the ground-breaking aspect being the exploration of decay rates of mRNAs.
  • The study revealed a collection of short-lived chondrocyte mRNAs presumed to be greatly responsive to altered transcriptional input, as well as mRNAs whose decay rates were affected in OA chondrocytes.

Cite This Article

APA
Tew SR, McDermott BT, Fentem RB, Peffers MJ, Clegg PD. (2014). Transcriptome-wide analysis of messenger RNA decay in normal and osteoarthritic human articular chondrocytes. Arthritis Rheumatol, 66(11), 3052-3061. https://doi.org/10.1002/art.38849

Publication

Arthritis & rheumatology (Hoboken, N.J.)
ISSN: 2326-5205
NlmUniqueID: 101623795
Country: United States
Language: English
Volume: 66
Issue: 11
Pages: 3052-3061

Researcher Affiliations

Tew, Simon R
  • University of Liverpool, Leahurst Campus, Neston, Cheshire, UK.
McDermott, Benjamin T
    Fentem, Rory B
      Peffers, Mandy J
        Clegg, Peter D

          MeSH Terms

          • Adolescent
          • Adult
          • Aged
          • Aged, 80 and over
          • Aging / metabolism
          • Animals
          • Cartilage, Articular / metabolism
          • Cartilage, Articular / pathology
          • Cells, Cultured
          • Chondrocytes / metabolism
          • Chondrocytes / pathology
          • Female
          • Gene Expression Profiling
          • Gene Expression Regulation
          • Horses
          • Humans
          • In Vitro Techniques
          • Male
          • Middle Aged
          • Models, Animal
          • Osteoarthritis, Knee / metabolism
          • Osteoarthritis, Knee / pathology
          • Osteoarthritis, Knee / physiopathology
          • RNA Stability / physiology
          • RNA, Messenger / metabolism
          • Young Adult

          Grant Funding

          • 19385 / Versus Arthritis
          • MR/K006312/1 / Medical Research Council
          • 19385 / Arthritis Research UK
          • WT088557MA / Wellcome Trust

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          Citations

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