TY - JOUR
T1 - Neural epidermal growth factor-like 1 protein (NELL-1) associated membranous nephropathy
AU - Sethi, Sanjeev
AU - Debiec, Hanna
AU - Madden, Benjamin
AU - Charlesworth, M. Cristine
AU - Morelle, Johann
AU - Gross, Lou Ann
AU - Ravindran, Aishwarya
AU - Buob, David
AU - Jadoul, Michel
AU - Fervenza, Fernando C.
AU - Ronco, Pierre
N1 - Funding Information:
We would like to thank the Mayo Clinic, Proteomics Core (a shared resource of the Mayo Clinic Cancer Center [National Cancer Institute {NCI} P30 CA15083]); Department of Laboratory Medicine and Pathology and the Pathology Research Core, Mayo Clinic; and the Fulk Family Foundation. PR received the following: European Research Council ERC-2012-ADG_20120314 grant no. 322947, 7th Framework Programme of the European Community contract no. 2012-305608 (European Consortium for High-Throughput Research in Rare Kidney Diseases), and the National Research Agency grant MNaims (ANR-17-CE17-0012-01). We are greatly indebted to the clinicians who took care of the patients in the Department of Nephrology and Dialysis at Tenon Hospital, Paris, over the last 20 years; to Isabelle Brocheriou (MD, PhD) from the Department of Pathology, Pitié Salpêtrière Hospital, Paris; and to Nadhir Yousfi (PhD) for help in Western blotting experiments. We would like to thank Romain Morichon (PhD), Confocal Microscopy Platform, Saint-Antoine Hospital, Paris; Eva Compérat; and all staff members of the Tenon Hospital Biological Resource Center (BRC CANCER HUEP- Paris) for their help in centralizing and managing biological data collection. We also thank the clinicians from the UC Louvain Kidney Disease Network for patients’ referral; Selda Aydin (MD, PhD) from the Department of Pathology of the Cliniques universitaires Saint-Luc, Brussels, for pathological diagnosis; and Nicolas Hanset (MD) for data collection.
Funding Information:
We would like to thank the Mayo Clinic, Proteomics Core (a shared resource of the Mayo Clinic Cancer Center [National Cancer Institute {NCI} P30 CA15083]); Department of Laboratory Medicine and Pathology and the Pathology Research Core, Mayo Clinic; and the Fulk Family Foundation. PR received the following: European Research Council ERC-2012-ADG_20120314 grant no. 322947, 7th Framework Programme of the European Community contract no. 2012-305608 (European Consortium for High-Throughput Research in Rare Kidney Diseases), and the National Research Agency grant MNaims (ANR-17-CE17-0012-01). We are greatly indebted to the clinicians who took care of the patients in the Department of Nephrology and Dialysis at Tenon Hospital, Paris, over the last 20 years; to Isabelle Brocheriou (MD, PhD) from the Department of Pathology, Piti? Salp?tri?re Hospital, Paris; and to Nadhir Yousfi (PhD) for help in Western blotting experiments. We would like to thank Romain Morichon (PhD), Confocal Microscopy Platform, Saint-Antoine Hospital, Paris; Eva Comp?rat; and all staff members of the Tenon Hospital Biological Resource Center (BRC CANCER HUEP- Paris) for their help in centralizing and managing biological data collection. We also thank the clinicians from the UC Louvain Kidney Disease Network for patients? referral; Selda Aydin (MD, PhD) from the Department of Pathology of the Cliniques universitaires Saint-Luc, Brussels, for pathological diagnosis; and Nicolas Hanset (MD) for data collection. SS and FCF designed the study. SS wrote the manuscript, interpreted the kidney biopsy, clinical, IHC and MS data. BM and CC performed the laser microdissection and MS. AR helped in gathering clinical data. LG performed the IHC. PR, DB, and HD provided tissue for the validation cohort and also performed the confocal studies and Western blot analysis. JM and MJ provided clinical information and tissue for the Belgian validation cohort. The manuscript was drafted and written by SS, with input as appropriate from the rest of the investigators.
Publisher Copyright:
© 2019 International Society of Nephrology
PY - 2020/1
Y1 - 2020/1
N2 - Membranous nephropathy is characterized by deposition of immune complexes along the glomerular basement membrane. PLA2R and THSD7A are target antigens in 70% and 1-5% of primary membranous nephropathy cases, respectively. In the remaining cases, the target antigen is unknown. Here, laser microdissection of glomeruli followed by mass spectrometry was used to identify novel antigen(s) in PLA2R-negative membranous nephropathy. An initial pilot mass spectrometry study in 35 cases of PLA2R-negative membranous nephropathy showed high spectral counts for neural tissue encoding protein with EGF-like repeats, NELL-1, in six cases. Mass spectrometry failed to detect NELL-1 in 23 PLA2R-associated membranous nephropathy and 88 controls. NELL-1 was localized by immunohistochemistry, which showed bright granular glomerular basement membrane staining for NELL-1 in all six cases. Next, an additional 23 NELL-1 positive cases of membranous nephropathy were identified by immunohistochemistry in a discovery cohort of 91 PLA2R-negative membranous nephropathy cases, 14 were confirmed by mass spectrometry. Thus, 29 of 126 PLA2R-negative cases were positive for NELL-1. PLA2R-associated membranous nephropathy and controls stained negative for NELL-1. We then identified five NELL-1 positive cases of membranous nephropathy out of 84 PLA2R and THSD7A-negative cases in two validation cohorts from France and Belgium. By confocal microscopy, both IgG and NELL-1 co-localized to the glomerular basement membrane. Western blot analysis showed reactivity to NELL-1 in five available sera, but no reactivity in control sera. Clinical and biopsy findings of NELL-1 positive membranous nephropathy showed features of primary membranous nephropathy. Thus, a subset of membranous nephropathy is associated with accumulation and co-localization of NELL-1 and IgG along the glomerular basement membrane, and with anti-NELL-1 antibodies in the serum. Hence, NELL-1 defines a distinct type of primary membranous nephropathy.
AB - Membranous nephropathy is characterized by deposition of immune complexes along the glomerular basement membrane. PLA2R and THSD7A are target antigens in 70% and 1-5% of primary membranous nephropathy cases, respectively. In the remaining cases, the target antigen is unknown. Here, laser microdissection of glomeruli followed by mass spectrometry was used to identify novel antigen(s) in PLA2R-negative membranous nephropathy. An initial pilot mass spectrometry study in 35 cases of PLA2R-negative membranous nephropathy showed high spectral counts for neural tissue encoding protein with EGF-like repeats, NELL-1, in six cases. Mass spectrometry failed to detect NELL-1 in 23 PLA2R-associated membranous nephropathy and 88 controls. NELL-1 was localized by immunohistochemistry, which showed bright granular glomerular basement membrane staining for NELL-1 in all six cases. Next, an additional 23 NELL-1 positive cases of membranous nephropathy were identified by immunohistochemistry in a discovery cohort of 91 PLA2R-negative membranous nephropathy cases, 14 were confirmed by mass spectrometry. Thus, 29 of 126 PLA2R-negative cases were positive for NELL-1. PLA2R-associated membranous nephropathy and controls stained negative for NELL-1. We then identified five NELL-1 positive cases of membranous nephropathy out of 84 PLA2R and THSD7A-negative cases in two validation cohorts from France and Belgium. By confocal microscopy, both IgG and NELL-1 co-localized to the glomerular basement membrane. Western blot analysis showed reactivity to NELL-1 in five available sera, but no reactivity in control sera. Clinical and biopsy findings of NELL-1 positive membranous nephropathy showed features of primary membranous nephropathy. Thus, a subset of membranous nephropathy is associated with accumulation and co-localization of NELL-1 and IgG along the glomerular basement membrane, and with anti-NELL-1 antibodies in the serum. Hence, NELL-1 defines a distinct type of primary membranous nephropathy.
KW - NELL-1
KW - mass spectrometry
KW - membranous nephropathy
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U2 - 10.1016/j.kint.2019.09.014
DO - 10.1016/j.kint.2019.09.014
M3 - Article
C2 - 31901340
AN - SCOPUS:85076263834
VL - 97
SP - 163
EP - 174
JO - Kidney International
JF - Kidney International
SN - 0085-2538
IS - 1
ER -