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Abstract: FR-PO956

Proteomics of Human Glomerulonephritis by Laser Microdissection and Liquid Chromatograph-Tandem Mass Spectrometry (LMD-LC MS/MS)

Session Information

Category: Pathology and Lab Medicine

  • 1601 Pathology and Lab Medicine: Basic

Authors

  • Kawata, Naoto, Showa University Fujigaoka Hospital, Yokohama, Japan
  • Kang, Dedong, Showa University, Tokyo, Japan
  • Inui, Kiyoko, Showa University Fujigaoka Hospital, Yokohama, Japan
  • Shibata, Takanori, Showa University, Tokyo, Japan
  • Yoshimura, Ashio, Shinyokohama Daiichi Clinic, Yokohama, Japan
  • Honda, Kazuho, Showa University, Tokyo, Japan
Background

Laser microdissection and liquid chromatograph tandem mass spectrometry (LMD-LC MS/MS) methods enable us to analyze the proteins from the tissue sections. In nephrology, they are preferentially applied in the diagnosis of abnormal protein deposition diseases, such as amyloidosis. In this study, we applied this method to the renal biopsy specimens obtained from the patients with IgA nephropathy(IgAN) and idiopatic membranous nephropathy(MN) to investigate its usefulness for the diagnosis and pathophysiological understanding of human glomerular diseases.

Methods

The 0.3mm2 of glomerular tissue were dissected by LMD from 10µm-thick sections of renal biopsy specimens obtained from five patients with IgA nephropathy, membranous nephropathy and kidney transplant donor, respectively. The samples were analyzed by LC-MS/MS and the results were investigated to clinical and histological findings.

Results

From the control glomeruli, more than 300 types of proteins such as cytoskeleton proteins, nuclear related proteins, Extracellular Matrix(ECM) related proteins and enzymes were identified. In addition to IgA1 and C3, IgAN showed significant increases in the factors related to oxidative stress and cell proliferation, such as heat shock protein 70/71/90, peroxiredoxin, and elongation factor 1α, as compared to controls. In MN, the proteins such as IgG1, IgG4, C3, C4a, and phospholipase-A2-Receptor(PLA2R) were significantly elevated compared to controls.

Conclusion

By applying LMD-LC MS/MS method to renal biopsy specimens, it was possible to identify the pathognomonic proteins for the diagnosis of IgAN and MN as well as immunohistochemistry. Furthermore, we identified the several factors involved in the pathogenesis of glomerular diseases. Although further investigation is necessary, it seems that the proteomics of glomerular proteins analyzed by LMD-LC MS/MS is a promising method for the diagnosis and pathophysiological understanding of human glomerulonephritis.