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Use of patient derived urine renal epithelial cells to confirm pathogenicity of PKHD1 alleles

Lookup NU author(s): Dr Elisa MolinariORCiD, Dr Shalabh Srivastava, Becky Dewhurst, Professor John SayerORCiD



This work is licensed under a Creative Commons Attribution 4.0 International License (CC BY 4.0).


© 2020, The Author(s).Background: PKHD1 is the main genetic cause of autosomal recessive polycystic kidney disease (ARPKD), a hereditary hepato-renal fibrocystic disorder which is the most important cause of end-stage renal disease during early childhood. ARPKD can also present in adulthood with milder phenotypes. In this study, we describe a 24-year-old woman with atypical polycystic kidney, no family history of renal disease and no obvious extra-renal manifestations who was referred for genetic investigation. Methods: We used a combination of next generation sequencing, Sanger sequencing and RNA and microscopy studies performed on urine-derived renal epithelial cells (URECs) to provide a genetic diagnosis of ARPKD. Results: A next generation sequencing panel of cystic ciliopathy genes allowed the identification of two heterozygous sequence changes in PKHD1 (c.6900C > T; p.(Asn2300=) and c.7964A > C; p.(His2655Pro)). The pathogenicity of the synonymous PKHD1 variant is not clear and requires RNA studies, which cannot be carried out efficiently on RNA extracted from proband blood, due to the low expression levels of PKHD1 in lymphocytes. Using URECs as a source of kidney-specific RNA, we show that PKHD1 is alternatively spliced around exon 43, both in control and proband URECs. The variant p.(Asn2300=) shifts the expression ratio in favour of a shorter, out-of-frame transcript. To further study the phenotypic consequence of these variants, we investigated the ciliary phenotype of patient URECs, which were abnormally elongated and presented multiple blebs along the axoneme. Conclusions: We confirm the power of URECs as a tool for functional studies on candidate variants in inherited renal disease, especially when the expression of the gene of interest is restricted to the kidney and we describe, for the first time, ciliary abnormalities in ARPKD patient cells.

Publication metadata

Author(s): Molinari E, Srivastava S, Dewhurst RM, Sayer JA

Publication type: Article

Publication status: Published

Journal: BMC Nephrology

Year: 2020

Volume: 21

Issue: 1

Online publication date: 15/10/2020

Acceptance date: 08/10/2020

Date deposited: 19/11/2020

ISSN (electronic): 1471-2369

Publisher: BioMed Central Ltd


DOI: 10.1186/s12882-020-02094-z

PubMed id: 33059616


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Funder referenceFunder name
RP_006_20180227Kidney Research UK (was National Kidney Research Fund)