title: Discovery of novel differentiation markers in the early stage of chondrogenesis by glycoform-focused reverse proteomics and genomics.
abstract: BACKGROUND
  Osteoarthritis (OA) is one of the most common chronic diseases among adults, especially the elderly, which is characterized by destruction of the articular cartilage. Despite affecting more than 100 million individuals all over the world, therapy is currently limited to treating pain, which is a principal symptom of OA. New approaches to the treatment of OA that induce regeneration and repair of cartilage are strongly needed.
  
  
  METHODS
  To discover potent markers for chondrogenic differentiation, glycoform-focused reverse proteomics and genomics were performed on the basis of glycoblotting-based comprehensive approach.
  
  
  RESULTS
  Expression levels of high-mannose type N-glycans were up-regulated significantly at the late stage of differentiation of the mouse chondroprogenitor cells. Among 246 glycoproteins carrying this glycotype identified by ConA affinity chromatography and LC/MS, it was demonstrated that 52% are classified as cell surface glycoproteins. Gene expression levels indicated that mRNAs for 15 glycoproteins increased distinctly in the earlier stages during differentiation compared with Type II collagen. The feasibility of mouse chondrocyte markers in human chondrogenesis model was demonstrated by testing gene expression levels of these 15 glycoproteins during differentiation in human mesenchymal stem cells.
  
  
  CONCLUSIONS
  The results showed clearly an evidence of up-regulation of 5 genes, ectonucleotide pyrophosphatase/phosphodiesterase family member 1, collagen alpha-1(III) chain, collagen alpha-1(XI) chain, aquaporin-1, and netrin receptor UNC5B, in the early stages of differentiation.
  
  
  CONCLUSIONS
  These cell surface 5 glycoproteins become highly sensitive differentiation markers of human chondrocytes that contribute to regenerative therapies, and development of novel therapeutic reagents.
authors.affiliation: Shionogi Innovation Center for Drug Discovery, Shionogi & Co. Ltd., Kita-ku, Sapporo 001-0021, Japan.
authors.forename: Takeshi
authors.initials: T
authors.lastname: Ishihara
authors.forename: Kiyoshi
authors.initials: K
authors.lastname: Kakiya
authors.forename: Koji
authors.initials: K
authors.lastname: Takahashi
authors.forename: Hiroto
authors.initials: H
authors.lastname: Miwa
authors.forename: Masatomo
authors.initials: M
authors.lastname: Rokushima
authors.forename: Tomoyo
authors.initials: T
authors.lastname: Yoshinaga
authors.forename: Yoshikazu
authors.initials: Y
authors.lastname: Tanaka
authors.forename: Takaomi
authors.initials: T
authors.lastname: Ito
authors.forename: Hiroko
authors.initials: H
authors.lastname: Togame
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Takemoto
authors.forename: Maho
authors.initials: M
authors.lastname: Amano
authors.forename: Norimasa
authors.initials: N
authors.lastname: Iwasaki
authors.forename: Akio
authors.initials: A
authors.lastname: Minami
authors.forename: Shin-Ichiro
authors.initials: S
authors.lastname: Nishimura
mesh_terms: D000328:Adult; D000818:Animals; D000943:Antigens, Differentiation / Q000378:metabolism*; D002454:Cell Differentiation; D002478:Cells, Cultured; D019902:Chondrocytes / Q000166:cytology* / Q000378:metabolism*; D020219:Chondrogenesis / Q000502:physiology*; D020869:Gene Expression Profiling; D023281:Genomics; D006023:Glycoproteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism; D051379:Mice; D020411:Oligonucleotide Array Sequence Analysis; D011134:Polysaccharides / Q000378:metabolism; D040901:Proteomics; D012333:RNA, Messenger / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D000943:Antigens, Differentiation; D006023:Glycoproteins; D011134:Polysaccharides; D012333:RNA, Messenger
keywords: Chondrogenic differentiation; Glycoform-focused reverse proteomics and genomics; Glycomics; Mass spectrometry; Osteoarthritis