Skip to main content

Advertisement

LC-MS/MS-based shotgun proteomics identified the targets of arthritis-related microRNA

microRNAs (miRNAs), which are class of post-transcriptional regulators such as short 19 to 23-nucleotide non-coding RNAs, complementarily bind seed sequences in the 3'-untranslational region of multiple target mRNAs, resulting in their suppression of translation or degradation [1]. In the former case, since the mRNA expression of the targets does not any change, transcriptomics approach, such as expression array, cannot identify the targets.

Recent studies shed light on the fine-tuning mechanism of miRNAs in myriad biological processes including development [2], tumorigenesis [3] and inflammation [4]. We have identified enhancement of mir-146a expression in rheumatoid arthritis synoviocyte and macrophages [5], whilst suppression of them in osteoarthritis [6]. Another group also have identified the enhancement of mir-146a and mir-155 in response to bacterial pathogen such as lipopolysaccaride [7]. Recently, mice lacking of mir-155 are resistant to collagen-induced arthritis (CIA) [8], whilst administration of mir-146a complexed with aterocollagen into joint attenuates pathological condition of CIA [9]. These results indicate that mir-146a and mir-155 plays an important role for developing arthritis and inflammation. However, the targets of both two miRNAs and their molecular mechanisms are not still fully identified.

In this study, in order to identify the targets of them in translational level, we established gain of function models using adenovirus- and CMV promoter-mediated overexpression in several culture models and performed liquid chromatography-tandem mass spectrometry-based shotgun proteomics in these models.

References

  1. 1.

    Lee RC, Feinbaum RL, Ambros V: The C. elegans heterochronic gene lin-4 encodes small RNAs with antisense complementarity to lin-14. Cell. 1993, 75: 843-854. 10.1016/0092-8674(93)90529-Y.

  2. 2.

    Reinhart BJ, Slack FJ, Basson M, Pasquinelli AE, Bettinger JC, Rougvie AE, Horvitz HR, Ruvkun G: The 21-nucleotide let-7 RNA regulates developmental timing in Caenorhabditis elegans. Nature. 2005, 403: 901-906.

  3. 3.

    Johnson SM, Grosshans H, Shingara J, Byrom M, Jarvis R, Cheng A, Labourier E, Reinert KL, Brown D, Slack FJ: RAS is regulated by the let-7 microRNA family. Cell. 2005, 120: 635-647. 10.1016/j.cell.2005.01.014.

  4. 4.

    Lu LF, Boldin MP, Chaudhry A, Lin LL, Taganov KD, Hanada T, Yoshimura A, Baltimore D, Rudensky AY: Function of miR-146a in controlling Treg cell-mediated regulation of Th1 responses. Cell. 2010, 142: 914-929. 10.1016/j.cell.2010.08.012.

  5. 5.

    Nakasa T, Miyaki S, Okubo A, Hashimoto M, Nishida K, Ochi M, Asahara H: Expression of microRNA-146 in rheumatoid arthritis synovial tissue. Arthritis Rheum. 2008, 58: 1284-1292. 10.1002/art.23429.

  6. 6.

    Yamasaki K, Nakasa T, Miyaki S, Ishikawa M, Deie M, Adachi N, Yasunaga Y, Asahara H, Ochi M: Expression of MicroRNA-146a in osteoarthritis cartilage. Arthritis Rheum. 2009, 60: 1035-1041. 10.1002/art.24404.

  7. 7.

    Taganov KD, Boldin MP, Chang KJ, Baltimore D: NF-kappaB-dependent induction of microRNA miR-146, an inhibitor targeted to signaling proteins of innate immune responses. Proc Natl Acad Sci USA. 2006, 103: 12481-12486. 10.1073/pnas.0605298103.

  8. 8.

    Kurowska-Stolarska M, Alivernini S, Ballantine LE, Asquith DL, Millar NL, Gilchrist DS, Reilly J, Ierna M, Fraser AR, Stolarski B, McSharry C, Hueber AJ, Baxter D, Hunter J, Gay S, Liew FY, McInnes IB: MicroRNA-155 as a proinflammatory regulator in clinical and experimental arthritis. Proc Natl Acad Sci USA. 2011, 108: 11193-11198. 10.1073/pnas.1019536108.

  9. 9.

    Nakasa T, Shibuya H, Nagata Y, Niimoto T, Ochi M: The inhibitory effect of microRNA-146a expression on bone destruction in collagen-induced arthritis. Arthritis Rheum. 2011, 63: 1582-1590. 10.1002/art.30321.

Download references

Acknowledgements

The authors sincerely thank Dr. Yanagiya R for helpful advice on preparation of adenovirus, and Dr. Inoue A for the gift of CMV vector for mir-146a.

Author information

Correspondence to Riho Kurata.

Rights and permissions

Reprints and Permissions

About this article

Keywords

  • Rheumatoid Arthritis
  • Arthritis
  • Osteoarthritis
  • Bacterial Pathogen
  • Multiple Target