Urinary Exosomal Cystatin C and Lipopolysaccharide Binding Protein as Biomarkers for Antibody−Mediated Rejection after Kidney Transplantation
Abstract
:1. Introduction
2. Materials and Methods
2.1. Patients
2.2. Histopathology
2.3. Urine Processing and Exosome Extraction
2.4. Sample Preparation for Proteome Analysis
2.5. Nano-LC-ESI-MS/MS Analysis
2.6. Database Searching and Label Free Quantitation
2.7. Biomarker Candidate Selection
2.8. Evaluation of Exosomes
2.9. Western Blot and Immunohistochemistry
2.10. Statistical Analysis
3. Results
3.1. Patient Characteristics
3.2. Urine Exosome Characterization and Exosomal Proteins
3.3. Discovery of Four Biomarkers for Discriminating Antibody-Mediated Rejection
3.4. Validation of the Four Biomarkers for Discriminating Antibody−Mediated Rejection
3.5. Comparison of LBP and CST3 Expression in Kidney Allograft Tissue According to Pathologic Groups
3.6. Levels of LBP, CST3, and RBP4 in the Urine and Serum
3.7. Protein–Protein Interaction Analysis of LBP and CST3
3.8. Combined Use of LBP and CST3 as Biomarkers for ABMR
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
Patents
References
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Discovery Cohort | Validation Cohort | ||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|
ABMR | BKVN | TCMR | NOMOA | DONOR | p | ABMR | TCMR | NOMOA | Control | p | |
(N = 12) | (N = 5) | (N = 8) | (N = 10) | (N = 24) | (N = 25) | (N = 10) | (N = 19) | (N = 25) | |||
Age (years) | 52.2 ± 13.6 | 47.2 ± 8.6 | 53.8 ± 12.7 | 48.4 ± 10.2 | 38.5 ± 7.9 | 0.002 | 46.0 ± 10.8 | 51.0 ± 10.9 | 51.0 ± 12.6 | 50.6 ± 9.7 | 0.37 |
Female sex | 5 (41.7) | 1 (20.0) | 1 (12.5) | 4 (40.0) | 13 (54.2) | 0.326 | 7 (28.0) | 3 (30.0) | 4 (21.1) | 13 (52.0) | 0.011 |
BMI (kg/m2) | 22.2 ± 2.6 | 23.7 ± 2.2 | 25.4 ± 4.3 | 21.6 ± 3.4 | 25.9 ± 3.8 | 0.011 | 22.0 ± 2.7 | 24.7 ± 2.7 | 22.7 ± 2.8 | 21.9 ± 3.0 | 0.351 |
Hypertension | 1 (8.3) | 0 | 2 (25.0) | 0 | 2 (8.0) | 0 | 1 (5.3) | 4 (16.0) | |||
Diabetes | 1 (8.3) | 1 (20.0) | 2 (25.0) | 4 (40.0) | 4 (16.0) | 5 (50.0) | 6 (31.6) | 3 (12.0) | |||
Glomerulonephritis | 1 (8.3) | 0 | 0 | 1 (10.0) | 5 (20.0) | 0 | 3 (15.8) | 6 (24.0) | |||
IgA nephropathy | 0 | 1 (20.0) | 1 (12.5) | 1 (10.0) | 3 (12.0) | 2 (20.0) | 1 (5.3) | 2 (8.0) | |||
FSGS | 0 | 0 | 0 | 1 (10.0) | 0 | 0 | 1 (5.3) | 0 | |||
PCKD | 0 | 0 | 1 (12.5) | 0 | 0 | 0 | 2 (10.5) | 1 (4.0) | |||
Unknown | 6 (50.0) | 3 (60.0) | 1 (12.5) | 3 (30.0) | 9 (36.0) | 2 (20.0) | 5 (26.3) | 8 (32.0) | |||
Others | 3 (25.0) | 1 (20.0) | 0 | 2 (20.0) | 2 (8.0) | 1 (10.0) | 0 | 1 (4.0) | |||
Serum creatinine (mg/dL) | 3.25 ± 1.41 | 4.37 ± 2.80 | 1.61 ± 0.38 | 2.33 ± 1.12 | 0.005 | 2.01 ± 1.08 | 2.45 ± 1.31 | 1.979 ± 0.75 | 0.508 | ||
Albumin: creatinine ratio | 1597 ± 1548 | 426.3 ± 643.3 | 290.7 ± 329.4 | 2816 ± 2170 | 0.059 | 1635 ± 1436 | 1422 ± 1737 | 732.6 ± 1027 | 0.236 | ||
Pre-emptive KT | 2 (16.7) | 1 (20.0) | 0 | 2 (20.0) | 0.594 | 2 (8.0) | 1 (10.0) | 1 (5.3) | 7 (28.0) | 0.05 | |
Pre-transplant dialysis (month) | 33.1 ± 40.2 | 30.0 ± 20.3 | 62.8 ± 71.6 | 48.4 ± 52.8 | 0.972 | 38.2 ± 54.0 | 49.4 ± 44.7 | 37.3 ± 44.3 | 36.2 ± 55.5 | 0.559 | |
KT to biopsy (month) | 182.8 ± 45.2 | 37.4 ± 62.1 | 64.3 ± 80.5 | 76.2 ± 67.9 | 0.003 | 105.3 ± 60.9 | 51.5 ± 59.1 | 68.9 ± 77.0 | 42.0 ± 36.5 | 0.002 | |
Induction | 0.025 | 0.046 | |||||||||
None | 10 (83.3) | 0 | 2 (25.0) | 3 (30.0) | 6 (24.0) | 2 (20.0) | 3 (15.8) | 1 (4.0) | |||
ATG | 1 (8.3) | 0 | 1 (12.5) | 2 (20.0) | 3 (12.0) | 1 (10.0) | 0 | 1 (4.0) | |||
Simulect | 1 (8.3) | 5 (100.0) | 5 (62.5) | 5 (50.0) | 13 (52.0) | 7 (70.0) | 15 (78.9) | 23 (92.0) | |||
Calcineurin inhibitor | 0.318 | 0.112 | |||||||||
Tacrolimus | 2 (16.7) | 5 (100.0) | 2 (25.0) | 5 (50.0) | 17 (68.0) | 4 (40.0) | 13 (68.4) | 19 (76.0) | |||
Cyclosporine | 10 (83.3) | 0 | 6 (75.0) | 5 (50.0) | 8 (32.0) | 6 (60.0) | 6 (31.6) | 6 (24.0) | |||
Steroid maintenance | 12 (100.0) | 5 (100.0) | 7 (87.5) | 8 (80.0) | 0.078 | 25 (100.0) | 10 (100.0) | 19 (100.0) | 24 (96.0) | 0.08 | |
A/B/DR HLA mismatch | 3.3 ± 1.5 | 3.2 ± 1.9 | 2.9 ± 2.0 | 3.1 ± 1.5 | 0.903 | 3.0 ± 1.5 | 3.7 ± 2.0 | 2.9 ± 1.6 | 3.0 ± 1.8 | 0.912 | |
ABO-incompatible KT | 1 (8.3) | 1 (20.0) | 0 | 0 | 0.241 | 3 (12.0) | 3 (30.0) | 5 (26.3) | 0 | 0.009 | |
DDKT | 2 (16.7) | 2 (40.0) | 3 (37.5) | 3 (30.0) | 0.077 | 8 (32.0) | 3 (30.0) | 6 (31.6) | 3 (12.0) | 0.41 | |
DSA detection | 6 (50) | 1 (20) | 3 (37.5) | 2 (25%) | 15 (60) | 4 (40) | 4 (21.05) | ||||
PRAI | 18.67 ± 29.68 | 1.800 ± 2.68 | 11.00 ± 13.85 | 7.625 ± 8.93 | 0.557 | 30.32 ± 31.49 | 30.60 ± 36.98 | 7.84 ± 20.06 | 0.063 | ||
PRAII | 43.42 ± 35.70 | 26.00 ± 35.80 | 41.63 ± 37.95 | 5.875 ± 15.82 | 0.144 | 65.32 ± 32.99 | 54.20 ± 39.77 | 25.16 ± 31.54 | 0.557 |
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Kim, M.J.; Lim, S.J.; Ko, Y.; Kwon, H.E.; Jung, J.H.; Kwon, H.; Go, H.; Park, Y.; Kim, T.-K.; Jung, M.; et al. Urinary Exosomal Cystatin C and Lipopolysaccharide Binding Protein as Biomarkers for Antibody−Mediated Rejection after Kidney Transplantation. Biomedicines 2022, 10, 2346. https://doi.org/10.3390/biomedicines10102346
Kim MJ, Lim SJ, Ko Y, Kwon HE, Jung JH, Kwon H, Go H, Park Y, Kim T-K, Jung M, et al. Urinary Exosomal Cystatin C and Lipopolysaccharide Binding Protein as Biomarkers for Antibody−Mediated Rejection after Kidney Transplantation. Biomedicines. 2022; 10(10):2346. https://doi.org/10.3390/biomedicines10102346
Chicago/Turabian StyleKim, Mi Joung, Seong Jun Lim, Youngmin Ko, Hye Eun Kwon, Joo Hee Jung, Hyunwook Kwon, Heounjeong Go, Yangsoon Park, Tae-Keun Kim, MinKyo Jung, and et al. 2022. "Urinary Exosomal Cystatin C and Lipopolysaccharide Binding Protein as Biomarkers for Antibody−Mediated Rejection after Kidney Transplantation" Biomedicines 10, no. 10: 2346. https://doi.org/10.3390/biomedicines10102346