Reduced ciliary polycystin-2 in induced pluripotent stem cells from polycystic kidney disease patients with PKD1 mutations

Benjamin S. Freedman, Albert Q. Lam, Jamie L. Sundsbak, Rossella Iatrino, Xuefeng Su, Sarah J. Koon, Maoqing Wu, Laurence Daheron, Peter C Harris, Jing Zhou, Joseph V. Bonventre

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Abstract

Heterozygous mutations in PKD1 or PKD2, which encode polycystin-1 (PC1) and polycystin-2(PC2), respectively, cause autosomal dominant PKD (ADPKD), whereas mutations in PKHD1, which encodes fibrocystin/polyductin (FPC), cause autosomal recessive PKD (ARPKD). However, the relationship between these proteins and the pathogenesis of PKD remains unclear. To model PKD in human cells, we established induced pluripotent stem (iPS) cell lines from fibroblasts of three ADPKD and two ARPKD patients. Genetic sequencing revealed unique heterozygous mutations in PKD1 of the parental ADPKD fibroblasts but no pathogenicmutations in PKD2. Undifferentiated PKD iPS cells, control iPS cells, and embryonic stem cells elaborated primary cilia and expressed PC1, PC2, and FPC at similar levels, and PKD and control iPS cells exhibited comparable rates of proliferation, apoptosis, and ciliogenesis. However, ADPKD iPS cells as well as somatic epithelial cells and hepatoblasts/biliary precursors differentiated fromthese cells expressed lower levels of PC2 at the cilium. Additional sequencing confirmed the retention of PKD1 heterozygous mutations in iPS cell lines from two patients but identified possible loss of heterozygosity in iPS cell lines from one patient. Furthermore, ectopic expression of wild-type PC1inADPKDiPS-derived hepatoblasts rescued ciliary PC2protein expression levels, and overexpression of PC1 but not a carboxy-terminal truncation mutant increased ciliary PC2 expression levels in mouse kidney cells. Taken together, these results suggest that PC1 regulates ciliary PC2 protein expression levels and support the use of PKD iPS cells for investigating disease pathophysiology.

Original languageEnglish (US)
Pages (from-to)1571-1586
Number of pages16
JournalJournal of the American Society of Nephrology
Volume24
Issue number10
DOIs
StatePublished - Oct 2013

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Induced Pluripotent Stem Cells
Polycystic Kidney Diseases
Mutation
Cilia
Cell Line
Fibroblasts
polycystic kidney disease 2 protein
Loss of Heterozygosity
Embryonic Stem Cells
Proteins
Epithelial Cells
Apoptosis
Kidney
polycystic kidney disease 1 protein

ASJC Scopus subject areas

  • Nephrology
  • Medicine(all)

Cite this

Reduced ciliary polycystin-2 in induced pluripotent stem cells from polycystic kidney disease patients with PKD1 mutations. / Freedman, Benjamin S.; Lam, Albert Q.; Sundsbak, Jamie L.; Iatrino, Rossella; Su, Xuefeng; Koon, Sarah J.; Wu, Maoqing; Daheron, Laurence; Harris, Peter C; Zhou, Jing; Bonventre, Joseph V.

In: Journal of the American Society of Nephrology, Vol. 24, No. 10, 10.2013, p. 1571-1586.

Research output: Contribution to journalArticle

Freedman, BS, Lam, AQ, Sundsbak, JL, Iatrino, R, Su, X, Koon, SJ, Wu, M, Daheron, L, Harris, PC, Zhou, J & Bonventre, JV 2013, 'Reduced ciliary polycystin-2 in induced pluripotent stem cells from polycystic kidney disease patients with PKD1 mutations', Journal of the American Society of Nephrology, vol. 24, no. 10, pp. 1571-1586. https://doi.org/10.1681/ASN.2012111089
Freedman, Benjamin S. ; Lam, Albert Q. ; Sundsbak, Jamie L. ; Iatrino, Rossella ; Su, Xuefeng ; Koon, Sarah J. ; Wu, Maoqing ; Daheron, Laurence ; Harris, Peter C ; Zhou, Jing ; Bonventre, Joseph V. / Reduced ciliary polycystin-2 in induced pluripotent stem cells from polycystic kidney disease patients with PKD1 mutations. In: Journal of the American Society of Nephrology. 2013 ; Vol. 24, No. 10. pp. 1571-1586.
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abstract = "Heterozygous mutations in PKD1 or PKD2, which encode polycystin-1 (PC1) and polycystin-2(PC2), respectively, cause autosomal dominant PKD (ADPKD), whereas mutations in PKHD1, which encodes fibrocystin/polyductin (FPC), cause autosomal recessive PKD (ARPKD). However, the relationship between these proteins and the pathogenesis of PKD remains unclear. To model PKD in human cells, we established induced pluripotent stem (iPS) cell lines from fibroblasts of three ADPKD and two ARPKD patients. Genetic sequencing revealed unique heterozygous mutations in PKD1 of the parental ADPKD fibroblasts but no pathogenicmutations in PKD2. Undifferentiated PKD iPS cells, control iPS cells, and embryonic stem cells elaborated primary cilia and expressed PC1, PC2, and FPC at similar levels, and PKD and control iPS cells exhibited comparable rates of proliferation, apoptosis, and ciliogenesis. However, ADPKD iPS cells as well as somatic epithelial cells and hepatoblasts/biliary precursors differentiated fromthese cells expressed lower levels of PC2 at the cilium. Additional sequencing confirmed the retention of PKD1 heterozygous mutations in iPS cell lines from two patients but identified possible loss of heterozygosity in iPS cell lines from one patient. Furthermore, ectopic expression of wild-type PC1inADPKDiPS-derived hepatoblasts rescued ciliary PC2protein expression levels, and overexpression of PC1 but not a carboxy-terminal truncation mutant increased ciliary PC2 expression levels in mouse kidney cells. Taken together, these results suggest that PC1 regulates ciliary PC2 protein expression levels and support the use of PKD iPS cells for investigating disease pathophysiology.",
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