RNA-seq analysis of clinical-grade osteochondral allografts reveals activation of early response genes

Yang Lin, Eric A. Lewallen, Emily T. Camilleri, Carolina A. Bonin, Dakota L. Jones, Amel Dudakovic, Catalina Galeano-Garces, Wei Wang, Marcel J. Karperien, Annalise N. Larson, Diane L. Dahm, Michael J. Stuart, Bruce A. Levy, Jay Smith, Daniel B. Ryssman, Jennifer J. Westendorf, Hee Jeong Im, Andre J. van Wijnen, Scott M. Riester, Aaron J. Krych

Research output: Contribution to journalArticle

18 Scopus citations

Abstract

Preservation of osteochondral allografts used for transplantation is critical to ensure favorable outcomes for patients after surgical treatment of cartilage defects. To study the biological effects of protocols currently used for cartilage storage, we investigated differences in gene expression between stored allograft cartilage and fresh cartilage from living donors using high throughput molecular screening strategies. We applied next generation RNA sequencing (RNA-seq) and real-time reverse transcription quantitative polymerase chain reaction (RT-qPCR) to assess genome-wide differences in mRNA expression between stored allograft cartilage and fresh cartilage tissue from living donors. Gene ontology analysis was used to characterize biological pathways associated with differentially expressed genes. Our studies establish reduced levels of mRNAs encoding cartilage related extracellular matrix (ECM) proteins (i.e., COL1A1, COL2A1, COL10A1, ACAN, DCN, HAPLN1, TNC, and COMP) in stored cartilage. These changes occur concomitantly with increased expression of “early response genes” that encode transcription factors mediating stress/cytoprotective responses (i.e., EGR1, EGR2, EGR3, MYC, FOS, FOSB, FOSL1, FOSL2, JUN, JUNB, and JUND). The elevated expression of “early response genes” and reduced levels of ECM-related mRNAs in stored cartilage allografts suggests that tissue viability may be maintained by a cytoprotective program that reduces cell metabolic activity. These findings have potential implications for future studies focused on quality assessment and clinical optimization of osteochondral allografts used for cartilage transplantation.

Original languageEnglish (US)
Pages (from-to)1950-1959
Number of pages10
JournalJournal of Orthopaedic Research
Volume34
Issue number11
DOIs
StatePublished - Nov 1 2016

Keywords

  • RNA sequencing
  • allograft
  • cartilage
  • osteochondral
  • transplant

ASJC Scopus subject areas

  • Orthopedics and Sports Medicine

Fingerprint Dive into the research topics of 'RNA-seq analysis of clinical-grade osteochondral allografts reveals activation of early response genes'. Together they form a unique fingerprint.

  • Cite this

    Lin, Y., Lewallen, E. A., Camilleri, E. T., Bonin, C. A., Jones, D. L., Dudakovic, A., Galeano-Garces, C., Wang, W., Karperien, M. J., Larson, A. N., Dahm, D. L., Stuart, M. J., Levy, B. A., Smith, J., Ryssman, D. B., Westendorf, J. J., Im, H. J., van Wijnen, A. J., Riester, S. M., & Krych, A. J. (2016). RNA-seq analysis of clinical-grade osteochondral allografts reveals activation of early response genes. Journal of Orthopaedic Research, 34(11), 1950-1959. https://doi.org/10.1002/jor.23209