Poster Presentation HUPO 2019 - 18th Human Proteome Organization World Congress

Glycomic and glycoproteomic approaches for development of novel glyco-biomarkers of cardiac fibrogenesis using a mouse model of dilated cardiomyopathy (#744)

Chiaki Nagai-Okatani 1 , Mitsuhiro Nishigori 2 , Takashi Sato 1 , Naoto Minamino 2 , Hiroyuki Kaji 1 , Atsushi Kuno 1
  1. Biotechnology Research Institute for Drug Discovery, National Institute of Advanced Industrial Science and Technology, Tsukuba, Japan
  2. Omics Research Center, National Cerebral and Cardiovascular Center, Suita, Japan

Cardiac fibrosis is a typical phenomenon in failing hearts for most cardiac diseases, including dilated cardiomyopathy (DCM), and its specific detection and quantification is crucial for the analysis of cardiac remodeling. Since cardiac fibrosis is characterized by extensive remodeling of the myocardial extracellular matrix (ECM), in which glycoproteins are major components, we assumed that fibrosis-related alterations in the cardiac glycome and glycoproteome would be suitable targets for the detection of cardiac fibrosis. In the present study1), we compared protein glycosylation between formalin-fixed paraffin-embedded heart sections of normal and DCM model mice by laser microdissection-assisted lectin microarray2,3). Among 45 lectins, Wisteria floribunda agglutinin (WFA) was selected as the most suitable lectin for staining cardiac fibrotic tissues. The extent of WFA staining was highly correlated (r > 0.98) with that of picrosirius red staining, a common collagen staining method. However, the fibrosis-specific WFA staining did not overlap with staining signals of major collagen fiber components, indicating that collagen fibers themselves would not be WFA ligands. Further histochemical analysis with N-glycosidase revealed that the fibrosis-specific WFA staining was attributable to the binding of WFA to N-glycoproteins. Using a mass spectrometry-based approach3), we identified WFA-binding N-glycoproteins expressed in DCM hearts, many of which were fibrogenesis-related ECM proteins, as expected. Additionally, the identified glycoproteins carrying WFA-binding N-glycans were detected only in DCM hearts, suggesting their cooperative glycosylation alterations with disease progression. Collectively, these results indicate that WFA staining is more suitable for the quantitative assessment of cardiac fibrogenic activity than current collagen staining methods. Furthermore, given that plasma WFA-binding glycoprotein levels were significantly correlated with the echocardiographic parameters for left ventricular remodeling, cardiac WFA-binding glycoproteins are candidate circulating glyco-biomarkers for the quantification and monitoring of cardiac fibrogenesis.

  1. Nagai-Okatani C et al. Lab Invest (in press).
  2. Zou X*, Yoshida M*, Nagai-Okatani C* et al. Sci Rep 7:43560 (2017).
  3. Narimatsu H et al. J Proteome Res 17:4097-4112 (2018).