Cilia–to–basement membrane signaling is a biomechanical driver in models of autosomal dominant polycystic kidney disease
Manal Mazloum, Brice Lapin, Rushdi Alghamdi, Jessica Vandensteen, Martine Burtin, Pascal Houillier, Lydie Cheval, Gilles Crambert, Vicky Scata, Camille Cohen, Christoph Schell, Michael Rehman, Amandine Aka, Karim Ourahmoun, Rui Benedito, E. Wolfgang Kuehn, Stéphanie Descroix, Tilman Busch, Michael Köttgen, Serge Garbay, Marie-Christine Verpont, Ellie Tang, Brigitte Lelongt, Nicolas Cagnard, Stefan Somlo, Sylvie Coscoy, Fabiola Terzi, Amandine Viau, Frank Bienaimé
Manal Mazloum, Brice Lapin, Rushdi Alghamdi, Jessica Vandensteen, Martine Burtin, Pascal Houillier, Lydie Cheval, Gilles Crambert, Vicky Scata, Camille Cohen, Christoph Schell, Michael Rehman, Amandine Aka, Karim Ourahmoun, Rui Benedito, E. Wolfgang Kuehn, Stéphanie Descroix, Tilman Busch, Michael Köttgen, Serge Garbay, Marie-Christine Verpont, Ellie Tang, Brigitte Lelongt, Nicolas Cagnard, Stefan Somlo, Sylvie Coscoy, Fabiola Terzi, Amandine Viau, Frank Bienaimé
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Research Article Genetics Nephrology

Cilia–to–basement membrane signaling is a biomechanical driver in models of autosomal dominant polycystic kidney disease

Abstract

Autosomal dominant polycystic kidney disease (ADPKD), the leading genetic cause of kidney failure, results from loss-of-function mutations in PKD1, encoding polycystin-1 (PC1). PC1 localizes to the primary cilium. In the absence of PC1, adverse signaling from the primary cilium orchestrates cyst formation, but the biomechanical underpinnings of this cilia-dependent cyst activation (CDCA) remain unclear. Combining tubule-specific orthologous mouse models with a tubule-on-chip platform, we show that PC1 and cilia govern the composition, mechanical properties, and shape of the tubular basement membrane (TBM), the principal rigid determinant of tubule geometry. PC1 loss triggered TBM thinning, heparan sulfate enrichment, and deformation, leading to distension, preferentially of the distal nephron. These changes were driven by a cilia-dependent transcriptional program, with GLIS2 — a key CDCA effector — participating as a downstream mediator. Reduction of TBM stiffness amplified Pkd1–/– tubule-on-chip dilation and increased cyst formation in vivo. Conversely, increasing luminal pressure through ureteral obstruction induced disproportionate distension of Pkd1-deficient tubules and triggered an irreversible cystogenic program. Together, these findings establish a TBM-centered biomechanical model of ADPKD in which tubule deformation is governed by both basolateral and luminal mechanical factors and identify the cilium/TBM axis, operating in part through GLIS2, as a central driver of cystogenesis.

Authors

Manal Mazloum, Brice Lapin, Rushdi Alghamdi, Jessica Vandensteen, Martine Burtin, Pascal Houillier, Lydie Cheval, Gilles Crambert, Vicky Scata, Camille Cohen, Christoph Schell, Michael Rehman, Amandine Aka, Karim Ourahmoun, Rui Benedito, E. Wolfgang Kuehn, Stéphanie Descroix, Tilman Busch, Michael Köttgen, Serge Garbay, Marie-Christine Verpont, Ellie Tang, Brigitte Lelongt, Nicolas Cagnard, Stefan Somlo, Sylvie Coscoy, Fabiola Terzi, Amandine Viau, Frank Bienaimé

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Figure 8

ECM mechanics modulate Pkd1–/– tubule-on-chip dilation in vitro.

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ECM mechanics modulate Pkd1–/– tubule-on-chip dilation in vitro. (A and ...
(A and B) Representative images of tubules-on-chip formed by parental or Pkd1–/– mIMCD3 clones in scaffolds containing 9.5 (A) or 6 (B) g/L collagen I, 1 and 11 days after confluency. Arrows indicate tubule diameter. Scale bars: 100 μm. (C) Quantification of the normalized tubule deformation in the corresponding tubules-on-chip at 11 days. Each dot represents 1 tubule from n = 8 for parental cells and n = 8, 5, and 4 chips for Pkd1–/– clones 1, 2, and 3, respectively. (D–F) Labeling (D) and quantification of Ki67 (E) and internuclear distance (F) in parental and Pkd1–/– tubules-on-chip. Each dot represents 1 tubule (parental cells: N = 3 chips/n = 9–11 tubules; Pkd1–/– cells: N = 3 chips/n = 9–12 tubules). Mann-Whitney test: **P < 0.01. Scale bar: 100 μm. The box-and-whisker plots depict the minimum and maximum values (whiskers), the upper and lower quartiles, and the median. (G) Representative orthogonal sections of ciliated Pkd1–/– tubules-on-chip embedded in 6 or 9.5 g/L collagen I and maintained under static conditions for 5 days. Cilia were identified by immunostaining of acetylated α-tubulin (green) and γ-tubulin (red). Scale bar: 100 μm. (H) Corresponding quantification of percentage of ciliated cells per tubule. Each dot represents 1 tubule (n = 3 tubules per condition). The reported P value was calculated using a Student’s t test. The data shown in this figure are derived from at least 2 different independent cell seeding experiments.