Primary URECs: a source to better understand the pathology of renal tubular epithelia in pediatric hereditary cystic kidney diseases

Orphanet Journal of Rare Diseases

Jens König¹  Kai Wohlgemuth¹  Mareike Dahmer-Heath¹  Carsten Bergmann²  Margarita E. Georgiadis³  Dieter Haffner³  Fatima Hassan³  Arne Mertens³  Amrit Khera³  Sarah Lüdiger³  Doris Franke³  Kathrin Swolana³  Wolfgang H. Ziegler³  Birga Soetje³  Metin Cetiner⁴  Max C. Liebau⁵  Claudia Dafinger⁵ 

[1] Department of General Pediatrics, University Children’s Hospital Münster;Department of Medicine IV, Faculty of Medicine and Medical Center, University of Freiburg;Department of Pediatric Kidney, Liver and Metabolic Diseases, Hannover Medical School;Department of Pediatric Nephrology, Pediatrics II, University of Duisburg-Essen;Department of Pediatrics and Center for Molecular Medicine, University Hospital Cologne and Faculty of Medicine, University of Cologne;

关键词: Urine-derived renal tubular epithelial cells (URECs);    Hereditary cystic kidney diseases;    Children;    3D culture;    Spheroids;    Epithelial morphogenesis;   

DOI  :  10.1186/s13023-022-02265-1

来源: DOAJ

【 摘 要 】

Abstract Background In pediatric hereditary cystic kidney diseases, epithelial cell defects mostly result from rare, autosomal recessively inherited pathogenic variants in genes encoding proteins of the cilia-centrosome complex. Consequences of individual gene variants on epithelial function are often difficult to predict and can furthermore depend on the patient’s genetic background. Here, we studied urine-derived renal tubular epithelial cells (URECs) from genetically determined, pediatric cohorts of different hereditary cystic kidney diseases, comprising autosomal recessive polycystic kidney disease, nephronophthisis (NPH) and the Bardet Biedl syndrome (BBS). UREC characteristics and behavior in epithelial function-related 3D cell culture were compared in order to identify gene and variant-specific properties and to determine aspects of epithelial (cell) dysfunction. Results UREC preparations from patients (19) and healthy controls (39) were studied in a qualitative and quantitative manner using primary cells cultured for up-to 21 days. In patients with biallelic pathogenic variants in PKHD1 or NPHP genes, we were able to receive satisfactory amounts of URECs of reproducible quality. In BBS patients, UREC yield was lower and more dependent on the individual genotype. In contrast, in UREC preparations derived from healthy controls, no predictable and satisfactory outcome could be established. Considering cell proliferation, tubular origin and epithelial properties in 2D/3D culture conditions, we observed distinct and reproducible epithelial properties of URECs. In particular, the cells from patients carrying PKHD1 variants were characterized by a high incidence of defective morphogenesis of monolayered spheroids—a property proposed to be suitable for corrective intervention. Furthermore, we explored different ways to generate reference cell lines for both—patients and healthy controls—in order to eliminate restrictions in cell number and availability of primary URECs. Conclusions Ex vivo 3D cell culture of primary URECs represents a valuable, non-invasive source to evaluate epithelial cell function in kidney diseases and as such helps to elucidate the functional consequences of rare genetic disorders. In combination with genetically defined control cell lines to be generated in the future, the cultivation of primary URECs could become a relevant tool for testing personalized treatment of epithelial dysfunction in patients with hereditary cystic kidney disease.

【 授权许可】

Unknown