TY - JOUR
T1 - Urine-Derived Kidney Progenitor Cells in Cystinosis
AU - Veys, Koenraad
AU - Berlingerio, Sante Princiero
AU - David, Dries
AU - Bondue, Tjessa
AU - Held, Katharina
AU - Reda, Ahmed
AU - van den Broek, Martijn
AU - Theunis, Koen
AU - Janssen, Mirian
AU - Cornelissen, Elisabeth
AU - Vriens, Joris
AU - Diomedi-Camassei, Francesca
AU - Gijsbers, Rik
AU - van den Heuvel, Lambertus
AU - Arcolino, Fanny O.
AU - Levtchenko, Elena
N1 - Funding Information: Funding: This research was funded by the Research Foundation—Flanders (F.W.O. Vlaanderen), grant number 1801110N, the Cystinosis Research Network (CRN), Cystinosis Ireland and KU Leuven (C1 Internal Funding grant 3M170322). KV, FOA and DD are funded by the Research Foundation— Flanders (F.W.O Vlaanderen), grants 11Y5216N, 12Q99171N and 1S22919N, respectively. Funding Information: Acknowledgments: We thank Louise Medaer and Maxime Smits for their assistance with experiments while replacing Dries David. We also thank Inge Bongaers, Sandra van Aerschot, Irina Thiry, Christiana Adebayo, Sem Tuerlings and Antonina Mikorska for their technical assistance. We acknowledge M. van Dyck (UZ Leuven) for the collection of urine samples and care for our cystinosis patients; P. Van den Berghe (KU Leuven) of the Cell and Tissue Imaging cluster/Cell Imaging Core (CIC), supported by Hercules AKUL/15/37_GOH1816N and FWO G.0929.15 for support with confocal microscopy; B. Bussolatti (University of Turin, Italy) for sharing human papilla kidney stem cells; R. Masereeuw (University of Utrecht, Utrecht, The Netherlands) and F Emma (Laboratory of Pathology and Metabolism, Università Bambino Gesù, Rome, Italy) for cystine measurements. Library preparation, sequencing and statistical data analysis were performed by VIB Nucleomics Core (www.nucleomics.be; accessed date 13 March 2022). The authors acknowledge the internal fund from KU Leuven (C1 grant 3M170322). E.L. is supported by the Research Foundation—Flanders (F.W.O Vlaanderen), grant 1801110N, the Cystinosis Research Network (CRN) and Cystinosis Ireland. K.V, F.O.A and D.D are funded by the Research Foundation—Flanders (F.W.O Vlaanderen), grants 11Y5216N, 12Q9917N and 1S22919N, respectively. Publisher Copyright: © 2022 by the authors. Licensee MDPI, Basel, Switzerland.
PY - 2022/4/1
Y1 - 2022/4/1
N2 - Nephropathic cystinosis is an inherited lysosomal storage disorder caused by pathogenic variants in the cystinosin (CTNS) gene and is characterized by the excessive shedding of proximal tubular epithelial cells (PTECs) and podocytes into urine, development of the renal Fanconi syndrome and end-stage kidney disease (ESKD). We hypothesized that in compensation for epithelial cell losses, cystinosis kidneys undertake a regenerative effort, and searched for the presence of kidney progenitor cells (KPCs) in the urine of cystinosis patients. Urine was cultured in a specific progenitor medium to isolate undifferentiated cells. Of these, clones were characterized by qPCR, subjected to a differentiation protocol to PTECs and podocytes and assessed by qPCR, Western blot, immunostainings and functional assays. Cystinosis patients voided high numbers of undifferentiated cells in urine, of which various clonal cell lines showed a high capacity for self-renewal and expressed kidney progenitor markers, which therefore were assigned as cystinosis urine-derived KPCs (Cys-uKPCs). Cys-uKPC clones showed the capacity to differentiate between functional PTECs and/or podocytes. Gene addition with wild-type CTNS using lentiviral vector technology resulted in significant reductions in cystine levels. We conclude that KPCs present in the urine of cystinosis patients can be isolated, differentiated and complemented with CTNS in vitro, serving as a novel tool for disease modeling.
AB - Nephropathic cystinosis is an inherited lysosomal storage disorder caused by pathogenic variants in the cystinosin (CTNS) gene and is characterized by the excessive shedding of proximal tubular epithelial cells (PTECs) and podocytes into urine, development of the renal Fanconi syndrome and end-stage kidney disease (ESKD). We hypothesized that in compensation for epithelial cell losses, cystinosis kidneys undertake a regenerative effort, and searched for the presence of kidney progenitor cells (KPCs) in the urine of cystinosis patients. Urine was cultured in a specific progenitor medium to isolate undifferentiated cells. Of these, clones were characterized by qPCR, subjected to a differentiation protocol to PTECs and podocytes and assessed by qPCR, Western blot, immunostainings and functional assays. Cystinosis patients voided high numbers of undifferentiated cells in urine, of which various clonal cell lines showed a high capacity for self-renewal and expressed kidney progenitor markers, which therefore were assigned as cystinosis urine-derived KPCs (Cys-uKPCs). Cys-uKPC clones showed the capacity to differentiate between functional PTECs and/or podocytes. Gene addition with wild-type CTNS using lentiviral vector technology resulted in significant reductions in cystine levels. We conclude that KPCs present in the urine of cystinosis patients can be isolated, differentiated and complemented with CTNS in vitro, serving as a novel tool for disease modeling.
KW - cell model
KW - cystinosis
KW - gene therapy
KW - kidney progenitors
UR - http://www.scopus.com/inward/record.url?scp=85127603188&partnerID=8YFLogxK
U2 - https://doi.org/10.3390/cells11071245
DO - https://doi.org/10.3390/cells11071245
M3 - Article
C2 - 35406807
SN - 2073-4409
VL - 11
JO - Cells
JF - Cells
IS - 7
M1 - 1245
ER -