Sergi Clotet-Freixas1, Caitriona M McEvoy2,3, Ihor Batruch4, Chiara Pastrello5, Max Kotlyar5, Julie Anh Dung Van2,6, Madhurangi Arambewela2, Alex Boshart2,6, Sofia Farkona2, Yun Niu5, Yanhong Li1, Olusegun Famure2, Andrea Bozovic7, Vathany Kulasingam7, Peixuen Chen2, S Joseph Kim2,3, Emilie Chan3, Sajad Moshkelgosha2,8, Syed Ashiqur Rahman9,10, Jishnu Das9,10, Tereza Martinu2,8,11, Stephen Juvet2,8,11, Igor Jurisica5,12,13,14, Andrzej Chruscinski2,11, Rohan John2,7, Ana Konvalinka1,3,6,7,11. 1. Toronto General Hospital Research Institute, University Health Network, Toronto, Ontario, Canada Sergi.ClotetFreixas@uhnresearch.ca Ana.Konvalinka@uhn.ca. 2. Toronto General Hospital Research Institute, University Health Network, Toronto, Ontario, Canada. 3. Division of Nephrology, Department of Medicine, University Health Network, Toronto, Ontario, Canada. 4. Department of Laboratory Medicine and Pathobiology, Lunenfeld-Tanenbaum Research Institute, Mount Sinai Hospital, Toronto, Ontario, Canada. 5. Krembil Research Institute, University Health Network, Toronto, Ontario, Canada. 6. Institute of Medical Science, University of Toronto, Toronto, Ontario, Canada. 7. Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, Ontario, Canada. 8. Division of Respirology, Toronto Lung Transplant Program, University Health Network, Toronto, Ontario, Canada. 9. Center for Systems Immunology, Department of Immunology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania. 10. Center for Systems Immunology, Department of Computational and Systems Biology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania. 11. Soham and Shaila Ajmera Family Transplant Centre, University Health Network, Toronto, Ontario, Canada. 12. Department of Medical Biophysics, University of Toronto, Toronto, Ontario, Canada. 13. Department of Computer Science, University of Toronto, Toronto, Ontario, Canada. 14. Institute of Neuroimmunology, Slovak Academy of Sciences, Bratislava, Slovakia.
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.
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.
Authors: Bernard M van den Berg; Gangqi Wang; Margien G S Boels; M Cristina Avramut; Erik Jansen; Wendy M P J Sol; Franck Lebrin; Anton Jan van Zonneveld; Eelco J P de Koning; Hans Vink; Hermann-Josef Gröne; Peter Carmeliet; Johan van der Vlag; Ton J Rabelink Journal: J Am Soc Nephrol Date: 2019-07-15 Impact factor: 10.121
Authors: T F Mueller; G Einecke; J Reeve; B Sis; M Mengel; G S Jhangri; S Bunnag; J Cruz; D Wishart; C Meng; G Broderick; B Kaplan; P F Halloran Journal: Am J Transplant Date: 2007-10-17 Impact factor: 8.086
Authors: Michael J Randles; Sophie Collinson; Tobias Starborg; Aleksandr Mironov; Mira Krendel; Eva Königshausen; Lorenz Sellin; Ian S D Roberts; Karl E Kadler; Jeffrey H Miner; Rachel Lennon Journal: Sci Rep Date: 2016-10-11 Impact factor: 4.379
Authors: S A Peeters; L Engelen; J Buijs; A Jorsal; H-H Parving; L Tarnow; P Rossing; C G Schalkwijk; C D A Stehouwer Journal: Cardiovasc Diabetol Date: 2017-04-26 Impact factor: 9.951
Authors: Ignacio M Seropian; Germán E González; Sebastián M Maller; Daniel H Berrocal; Antonio Abbate; Gabriel A Rabinovich Journal: Mediators Inflamm Date: 2018-11-05 Impact factor: 4.711
Authors: Sara Rahmati; Mark Abovsky; Chiara Pastrello; Max Kotlyar; Richard Lu; Christian A Cumbaa; Proton Rahman; Vinod Chandran; Igor Jurisica Journal: Nucleic Acids Res Date: 2020-01-08 Impact factor: 16.971
Authors: Timothy D Cummins; Erik A Korte; Sagar Bhayana; Michael L Merchant; Michelle T Barati; William E Smoyer; Jon B Klein Journal: Pediatr Nephrol Date: 2022-02-26 Impact factor: 3.651
Authors: Michael Randles; Franziska Lausecker; Qing Kong; Hani Suleiman; Graeme Reid; Maria Kolatsi-Joannou; Pinyuan Tian; Sara Falcone; Bernard Davenport; Paul Potter; Tom Van Agtmael; Jill Norman; David Long; Martin Humphries; Jeffrey Miner; Rachel Lennon Journal: J Am Soc Nephrol Date: 2021-05-28 Impact factor: 14.978