RT Journal Article
SR Electronic
T1 Proteomics Reveals Extracellular Matrix Injury in the Glomeruli and Tubulointerstitium of Kidney Allografts with Early Antibody-Mediated Rejection
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2020.03.03.975672
DO 10.1101/2020.03.03.975672
A1 Sergi Clotet-Freixas
A1 Caitriona M. McEvoy
A1 Ihor Batruch
A1 Julie Van
A1 Chiara Pastrello
A1 Max Kotlyar
A1 Madhurangi Arambewela
A1 Alex Boshart
A1 Sofia Farkona
A1 Yun Niu
A1 Yanhong Li
A1 Olusegun Famure
A1 Andrea Bozovic
A1 Vathany Kulasingam
A1 Peixuen Chen
A1 Joseph Kim
A1 Emilie Chan
A1 Sajad Moshkelgosha
A1 Tereza Martinu
A1 Stephen Juvet
A1 Igor Jurisica
A1 Andrzej Chruscinski
A1 Rohan John
A1 Ana Konvalinka
YR 2020
UL http://biorxiv.org/content/early/2020/03/05/2020.03.03.975672.abstract
AB Antibody-mediated rejection (AMR) accounts for >50% of kidney allograft losses. AMR is caused by donor-specific antibodies (DSA) against HLA and non-HLA antigens in the glomeruli and the tubulointerstitium, which together with inflammatory cytokines such as tumor necrosis factor alpha (TNFα) and interferon gamma (IFNɣ), trigger graft injury. Unfortunately, the mechanisms governing cell-specific injury in AMR remain unclear. We studied 30 for-cause kidney biopsies with early AMR, acute cellular rejection or acute tubular necrosis (‘non-AMR’). We laser-captured and microdissected glomeruli and tubulointerstitium, and subjected them to unbiased proteome analysis. 120/2026 glomerular and 180/2399 tubulointerstitial proteins were significantly differentially expressed in AMR vs. non-AMR biopsies (P<0.05). Basement membrane and extracellular matrix (ECM) proteins were significantly decreased in both AMR compartments. We verified decreased glomerular and tubulointerstitial LAMC1 expression, and decreased glomerular NPHS1 and PTPRO expression in AMR. Cathepsin-V (CTSV) was predicted to cleave ECM-proteins in the AMR glomeruli, and CTSL, CTSS and LGMN in the tubulointerstitium. We identified galectin-1, an immunomodulatory protein upregulated in the AMR glomeruli and linked to the ECM. Anti-HLA class-I antibodies significantly increased CTSV expression, and galectin-1 expression and secretion, in human glomerular endothelial cells. Glutathione S-transferase omega-1 (GSTO1), an ECM-modifying enzyme, was significantly increased in the AMR tubulointerstitium, and in TNFα-treated proximal tubular epithelial cells. IFNɣ and TNFα significantly increased CTSS and LGMN expression in these cells. Basement membranes are often remodeled in chronic AMR, and we demonstrated that this remodeling begins early in glomeruli and tubulointerstitium. Targeting ECM-remodeling in AMR may represent a new therapeutic opportunity.SIGNIFICANCE STATEMENT Antibody-mediated rejection (AMR) accounts for >50% of kidney allograft loss, and is caused by donor-specific antibodies against HLA antigens, which induce maladaptive responses in the kidney glomeruli and tubulointerstitium. This is the first unbiased proteomics analysis of laser-captured/microdissected glomeruli and tubulointerstitium from 30 indication kidney biopsies with early AMR, acute cellular rejection or acute tubular necrosis. >2,000 proteins were quantified in each compartment. We discovered that basement membrane and extracellular matrix (ECM) proteins were significantly decreased in both AMR compartments. Two ECM-modifying proteins, LGALS1 and GSTO1, were significantly increased in glomeruli and tubulointerstitium, respectively. LGALS1 and GSTO1 were upregulated by anti-HLA antibodies or AMR-related cytokines in primary kidney cells, and may represent therapeutic targets to ameliorate ECM-remodeling in AMR.ACRAcute cellular rejectionAMRAntibody-mediated rejectionATNAcute tubular necrosisBMBasement membraneCTSLCathepsin-LCTSSCathepsin-SCTSVCathepsin-VDSADonor-specific antibodiesECMExtracellular matrixELISAEnzyme-linked immunosorbent assayFDRFalse discovery rateGSTO1Glutathione S-transferase omega-1HGMECHuman glomerular microvascular endothelial cellsHLAHuman leucocyte antigenIFNɣInterferon gammaLAMC1Laminin subunit gamma-1LFQLabel-free quantificationLGALS1Galectin-1LGMNLegumainMS/MSTandem mass spectrometryNPHS1NephrinPTECProximal tubular epithelial cellsPTPROReceptor-type tyrosine-protein phosphatase OTGTransplant glomerulopathyTNFαTumor necrosis factor-alpha