Extracellular Matrix Injury of Kidney Allografts in Antibody-Mediated Rejection: A Proteomics Study

J Am Soc Nephrol. 2020 Nov;31(11):2705-2724. doi: 10.1681/ASN.2020030286. Epub 2020 Sep 8.

Abstract

Background: Antibody-mediated rejection (AMR) accounts for >50% of kidney allograft loss. Donor-specific antibodies (DSA) against HLA and non-HLA antigens in the glomeruli and the tubulointerstitium cause AMR while inflammatory cytokines such as TNFα trigger graft injury. The mechanisms governing cell-specific injury in AMR remain unclear.

Methods: Unbiased proteomic analysis of laser-captured and microdissected glomeruli and tubulointerstitium was performed on 30 for-cause kidney biopsy specimens with early AMR, acute cellular rejection (ACR), or acute tubular necrosis (ATN).

Results: A total of 107 of 2026 glomerular and 112 of 2399 tubulointerstitial proteins was significantly differentially expressed in AMR versus ACR; 112 of 2026 glomerular and 181 of 2399 tubulointerstitial proteins were significantly dysregulated in AMR versus ATN (P<0.05). Basement membrane and extracellular matrix (ECM) proteins were significantly decreased in both AMR compartments. Glomerular and tubulointerstitial laminin subunit γ-1 (LAMC1) expression decreased in AMR, as did glomerular nephrin (NPHS1) and receptor-type tyrosine-phosphatase O (PTPRO). The proteomic analysis revealed upregulated galectin-1, which is an immunomodulatory protein linked to the ECM, in AMR glomeruli. Anti-HLA class I antibodies significantly increased cathepsin-V (CTSV) expression and galectin-1 expression and secretion in human glomerular endothelial cells. CTSV had been predicted to cleave ECM proteins in the AMR glomeruli. Glutathione S-transferase ω-1, an ECM-modifying enzyme, was significantly increased in the AMR tubulointerstitium and in TNFα-treated proximal tubular epithelial cells.

Conclusions: Basement membranes are often remodeled in chronic AMR. Proteomic analysis performed on laser-captured and microdissected glomeruli and tubulointerstitium identified early ECM remodeling, which may represent a new therapeutic opportunity.

Keywords: acute allograft rejection; extracellular matrix; glomerular endothelial cells; kidney transplantation; proximal tubule; renal biopsy.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Aged
  • Allografts / metabolism
  • Allografts / pathology
  • Antibodies / metabolism
  • Basement Membrane / metabolism*
  • Biopsy
  • Cathepsins / metabolism
  • Cell Line
  • Cysteine Endopeptidases / metabolism
  • Extracellular Matrix / metabolism*
  • Extracellular Matrix / pathology
  • Female
  • Galectin 1 / genetics
  • Galectin 1 / metabolism
  • Gene Expression
  • Glutathione Transferase / metabolism
  • Graft Rejection / genetics
  • Graft Rejection / metabolism*
  • Graft Rejection / pathology*
  • Histocompatibility Antigens Class I / immunology
  • Humans
  • Kidney Glomerulus / metabolism
  • Kidney Glomerulus / pathology*
  • Kidney Transplantation
  • Kidney Tubules / metabolism
  • Kidney Tubules / pathology*
  • Laminin / metabolism
  • Male
  • Matrix Metalloproteinase 2 / metabolism
  • Matrix Metalloproteinase 3 / metabolism
  • Membrane Proteins / metabolism
  • Middle Aged
  • Necrosis
  • Proteomics
  • Receptor-Like Protein Tyrosine Phosphatases, Class 3 / metabolism
  • Tumor Necrosis Factor-alpha / pharmacology

Substances

  • Antibodies
  • Galectin 1
  • Histocompatibility Antigens Class I
  • LGALS1 protein, human
  • Laminin
  • Membrane Proteins
  • Tumor Necrosis Factor-alpha
  • laminin gamma 1
  • nephrin
  • GSTO1 protein, human
  • Glutathione Transferase
  • PTPRO protein, human
  • Receptor-Like Protein Tyrosine Phosphatases, Class 3
  • Cathepsins
  • Cysteine Endopeptidases
  • CTSV protein, human
  • MMP3 protein, human
  • Matrix Metalloproteinase 3
  • MMP2 protein, human
  • Matrix Metalloproteinase 2

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