title: RNA-seq analysis of clinical-grade osteochondral allografts reveals activation of early response genes.
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. © 2016 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 34:1950-1959, 2016.
authors.affiliation: Department of Orthopedic Surgery, Mayo Clinic, 200 First Street SW, Rochester, Minnesota, 55905.|Department of Orthopedic Surgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, P. R. China.
authors.forename: Yang
authors.initials: Y
authors.lastname: Lin
authors.affiliation: Department of Orthopedic Surgery, Mayo Clinic, 200 First Street SW, Rochester, Minnesota, 55905.
authors.forename: Eric A
authors.initials: EA
authors.lastname: Lewallen
authors.affiliation: Department of Orthopedic Surgery, Mayo Clinic, 200 First Street SW, Rochester, Minnesota, 55905.
authors.forename: Emily T
authors.initials: ET
authors.lastname: Camilleri
authors.affiliation: Department of Orthopedic Surgery, Mayo Clinic, 200 First Street SW, Rochester, Minnesota, 55905.
authors.forename: Carolina A
authors.initials: CA
authors.lastname: Bonin
authors.affiliation: Department of Orthopedic Surgery, Mayo Clinic, 200 First Street SW, Rochester, Minnesota, 55905.
authors.forename: Dakota L
authors.initials: DL
authors.lastname: Jones
authors.affiliation: Department of Orthopedic Surgery, Mayo Clinic, 200 First Street SW, Rochester, Minnesota, 55905.
authors.forename: Amel
authors.initials: A
authors.lastname: Dudakovic
authors.affiliation: Department of Orthopedic Surgery, Mayo Clinic, 200 First Street SW, Rochester, Minnesota, 55905.
authors.forename: Catalina
authors.initials: C
authors.lastname: Galeano-Garces
authors.affiliation: Department of Orthopedic Surgery, Mayo Clinic, 200 First Street SW, Rochester, Minnesota, 55905.|Department of Orthopedics, WuHan Orthopedics Hospital/Puai Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430033, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Wang
authors.affiliation: Department of Developmental BioEngineering, University of Twente, Enschede, The Netherlands.
authors.forename: Marcel J
authors.initials: MJ
authors.lastname: Karperien
authors.affiliation: Department of Orthopedic Surgery, Mayo Clinic, 200 First Street SW, Rochester, Minnesota, 55905.
authors.forename: Annalise N
authors.initials: AN
authors.lastname: Larson
authors.affiliation: Department of Orthopedic Surgery, Mayo Clinic, 200 First Street SW, Rochester, Minnesota, 55905.
authors.forename: Diane L
authors.initials: DL
authors.lastname: Dahm
authors.affiliation: Department of Orthopedic Surgery, Mayo Clinic, 200 First Street SW, Rochester, Minnesota, 55905.
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Stuart
authors.affiliation: Department of Orthopedic Surgery, Mayo Clinic, 200 First Street SW, Rochester, Minnesota, 55905.
authors.forename: Bruce A
authors.initials: BA
authors.lastname: Levy
authors.affiliation: Department of Orthopedic Surgery, Mayo Clinic, 200 First Street SW, Rochester, Minnesota, 55905.
authors.forename: Jay
authors.initials: J
authors.lastname: Smith
authors.affiliation: Department of Orthopedic Surgery, Mayo Clinic, 200 First Street SW, Rochester, Minnesota, 55905.
authors.forename: Daniel B
authors.initials: DB
authors.lastname: Ryssman
authors.affiliation: Department of Orthopedic Surgery, Mayo Clinic, 200 First Street SW, Rochester, Minnesota, 55905.
authors.forename: Jennifer J
authors.initials: JJ
authors.lastname: Westendorf
authors.affiliation: Department of Biochemistry, Rush University Medical Center, Chicago, Illinois, 60612.
authors.forename: Hee-Jeong
authors.initials: HJ
authors.lastname: Im
authors.affiliation: Department of Orthopedic Surgery, Mayo Clinic, 200 First Street SW, Rochester, Minnesota, 55905.
authors.forename: Andre J
authors.initials: AJ
authors.lastname: van Wijnen
authors.affiliation: Department of Orthopedic Surgery, Mayo Clinic, 200 First Street SW, Rochester, Minnesota, 55905.
authors.forename: Scott M
authors.initials: SM
authors.lastname: Riester
authors.affiliation: Department of Orthopedic Surgery, Mayo Clinic, 200 First Street SW, Rochester, Minnesota, 55905.
authors.forename: Aaron J
authors.initials: AJ
authors.lastname: Krych
mesh_terms: D002356:Cartilage / Q000378:metabolism*; D064595:Composite Tissue Allografts; D005109:Extracellular Matrix / Q000378:metabolism*; D015870:Gene Expression; D020869:Gene Expression Profiling; D006801:Humans; D017423:Sequence Analysis, RNA; D013312:Stress, Physiological; D014021:Tissue Preservation
keywords: RNA sequencing; allograft; cartilage; osteochondral; transplant