Acute Tubular Injury Is an Important Component in Type I Acute Antibody-Mediated Rejection

R, Johnson; Sarmarapungavan, D.; Parasuraman, Ravi; Maine, Gabriel N.; Rooney, Michelle Nayahamui; Wolforth, Stacy C.; Reddy, Gopal; Cohn, Steven; Dumler, Francis; Rocher, Leslie L.; Li, W.; Zhang, P.L.

doi:10.1016/j.transproceed.2013.05.012

Cited by 6 publications

(5 citation statements)

References 15 publications

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“…Using a laser-capture microdissection system (Leica), the glomeruli and tubulointerstitium were then isolated and collected in a tube with 35μL of protein extraction buffer containing 10mM Tris, 1mM EDTA, and 0.002% Zwittergent in MSgrade pure water (Sigma). To obtain comparable protein amounts across all samples, in each biopsy section, we standardized the amount of captured glomeruli and tubulointerstitium to 350,000μm 2 . In each biopsy section, this area included an average of 23 captured glomeruli (glomerular compartment) or 4-5 captured tubulointerstitial areas (tubulointerstitial compartment).The total protein amount per sample was below the limit of detection of standard protein quantification assays.…”

Section: Laser-capture Microdissection and Sample Preparation For Promentioning

confidence: 99%

“…ATN indicates tubular cell injury and is a component of AMR. 2 Chronic AMR is associated with an aggressive lesion, transplant glomerulopathy (TG), which portends poor prognosis. 3 Multilayering of peritubular capillary basement membranes (BMs) and duplication of glomerular BM differentiate TG from other forms of AMR; these abnormalities likely arise from repeated cycles of endothelial cell injury.…”

Section: Introductionmentioning

confidence: 99%

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Proteomics Reveals Extracellular Matrix Injury in the Glomeruli and Tubulointerstitium of Kidney Allografts with Early Antibody-Mediated Rejection

Clotet-Freixas

McEvoy

Batruch

et al. 2020

Preprint

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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. 3 SIGNIFICANCE STATEMENTAntibody-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 lasercaptured/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 ECMremodeling in AMR.with gene expression alterations. 27 However, compartment-specific molecular alterations, or changes in proteome composition/expression are unknown. To address this gap, we conducted a ...

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Section: Laser-capture Microdissection and Sample Preparation For Promentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Proteomics Reveals Extracellular Matrix Injury in the Glomeruli and Tubulointerstitium of Kidney Allografts with Early Antibody-Mediated Rejection

Clotet-Freixas

McEvoy

Batruch

et al. 2020

Preprint

View full text Add to dashboard Cite

show abstract

“…ATN indicates tubular cell injury and may be observed in the context of AMR. 2 Chronic AMR is associated with an aggressive lesion, transplant glomerulopathy (TG), which portends poor prognosis. 3 Multilayering of peritubular capillary basement membranes (BMs) may be seen in TG, together with the duplication of glomerular BMs; these abnormalities likely arise from repeated cycles of endothelial cell injury.…”

mentioning

confidence: 99%

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

Clotet-Freixas

McEvoy

Batruch

et al. 2020

JASN

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BackgroundAntibody-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.MethodsUnbiased 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).ResultsA 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.ConclusionsBasement 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.

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“…5,6 However, inflammation and injury to the proximal tubule also contributes to disease pathogenesis in many kidney disorders. [7][8][9][10] Considering that the tubulointerstitium accounts for >90% of kidney volume and that tubulointerstitial disease may occur independent of glomerular disease, 11 it is critical that urinary protein assessment in preeclampsia consider both glomerular and nonglomerular patterns of injury.…”

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confidence: 99%

Complement Activation and Kidney Injury Molecule-1–Associated Proximal Tubule Injury in Severe Preeclampsia

et al. 2014

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We enrolled 25 cases with severe preeclampsia, 25 controls with chronic hypertension, and 25 healthy controls without hypertension from a cohort of women receiving care at Brigham and Women's Hospital from March 2012 through March 2013. Institutional review board approval was obtained through the Partners Human Research Committee, and subjects gave informed consent. All procedures followed were in accordance with institutional guidelines. Methods Abstract-Kidney injury with proteinuria is a characteristic feature of preeclampsia, yet the nature of injury in specific regions of the nephron is incompletely understood. Our study aimed to use existing urinary biomarkers to describe the pattern of kidney injury and proteinuria in pregnancies affected by severe preeclampsia. We performed a case-control study of pregnant women from Brigham and Women's Hospital from 2012 to 2013. We matched cases of severe preeclampsia (n=25) 1:1 by parity and gestational age to 2 control groups with and without chronic hypertension. Urinary levels of kidney injury molecule-1 and complement components (C3a, C5a, and C5b-9) were measured by enzyme-linked immunosorbent assay, and other markers (albumin, β2 microglobulin, cystatin C, epithelial growth factor, neutrophil gelatinase-associated lipocalin, osteopontin, and uromodulin) were measured simultaneously with a multiplex electrochemiluminescence assay. Median values between groups were compared with the Wilcoxon signed-rank test and correlations with Spearman correlation coefficient. Analysis of urinary markers revealed higher excretion of albumin and kidney injury molecule-1 and lower excretion of neutrophil gelatinase-associated lipocalin and epithelial growth factor in severe preeclampsia compared with chronic hypertension and healthy controls. Among subjects with severe preeclampsia, urinary excretion of complement activation products correlated most closely with kidney injury molecule-1, a specific marker of proximal tubule injury (C5a: r=0.60; P=0.001; and C5b-9: r=0.75; P<0.0001). Taken together, we describe a pattern of kidney injury in severe preeclampsia that is characterized by glomerular impairment and complement-mediated inflammation and injury, possibly localized to the proximal tubule in

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Acute Tubular Injury Is an Important Component in Type I Acute Antibody-Mediated Rejection

Cited by 6 publications

References 15 publications

Proteomics Reveals Extracellular Matrix Injury in the Glomeruli and Tubulointerstitium of Kidney Allografts with Early Antibody-Mediated Rejection

Proteomics Reveals Extracellular Matrix Injury in the Glomeruli and Tubulointerstitium of Kidney Allografts with Early Antibody-Mediated Rejection

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

Complement Activation and Kidney Injury Molecule-1–Associated Proximal Tubule Injury in Severe Preeclampsia

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