NHA2 promotes cyst development in an in vitro model of polycystic kidney disease

Abstract

Key points Significant and selective up‐regulation of the Na+/H+ exchanger NHA2 (SLC9B2) was observed in cysts of patients with autosomal dominant polycystic kidney disease. Using the MDCK cell model of cystogenesis, it was found that NHA2 increases cyst size. Silencing or pharmacological inhibition of NHA2 inhibits cyst formation in vitro. Polycystin‐1 represses NHA2 expression via Ca2+/NFAT signalling whereas the dominant negative membrane‐anchored C‐terminal fragment (PC1‐MAT) increased NHA2 levels. Drugs (caffeine, theophylline) and hormones (vasopressin, aldosterone) known to exacerbate cysts elicit NHA2 expression. Taken together, the findings reveal NHA2 as a potential new player in salt and water homeostasis in the kidney and in the pathogenesis of polycystic kidney disease. Abstract Autosomal dominant polycystic kidney disease (ADPKD) is caused by mutations in PKD1 and PKD2 encoding polycystin‐1 (PC1) and polycystin‐2 (PC2), respectively. The molecular pathways linking polycystins to cyst development in ADPKD are still unclear. Intracystic fluid secretion via ion transporters and channels plays a crucial role in cyst expansion in ADPKD. Unexpectedly, we observed significant and selective up‐regulation of NHA2, a member of the SLC9B family of Na+/H+ exchangers, that correlated with cyst size and disease severity in ADPKD patients. Using three‐dimensional cultures of MDCK cells to model cystogenesis in vitro, we showed that ectopic expression of NHA2 is causal to increased cyst size. Induction of PC1 in MDCK cells inhibited NHA2 expression with concordant inhibition of Ca2+ influx through store‐dependent and ‐independent pathways, whereas reciprocal activation of Ca2+ influx by the dominant negative membrane‐anchored C‐terminal tail fragment of PC1 elevated NHA2. We showed that NHA2 is a target of Ca2+/NFAT signalling and is transcriptionally induced by methylxanthine drugs such as caffeine and theophylline, which are contraindicated in ADPKD patients. Finally, we observed robust induction of NHA2 by vasopressin, which is physiologically consistent with increased levels of circulating vasopressin and up‐regulation of vasopressin V2 receptors in ADPKD. Our findings have mechanistic implications on the emerging use of vasopressin V2 receptor antagonists such as tolvaptan as safe and effective therapy for polycystic kidney disease and reveal a potential new regulator of transepithelial salt and water transport in the kidney.

Chemical and molecular bases of dome formation in human colorectal cancer cells mediated by sulphur compounds from Cucumis melo var. conomon

“…In addition, the domes are reportedly formed in response to known differentiation inducers [14,15]. Thus, to date, dome formation has been used as a morphological marker in the cell cultures for the onset of cell differentiation for studying transepithelial transport and cell polarity, because that unidirectional fluid movement mimics the active transepithelial transport seen in differentiated epithelial tissues of colon and kidney [10,11,[16][17][18].…”

A new coupling of an acid‐base transporter to PKD and cyst formation

“…Prasad et al . () present strong evidence for the involvement of NHA2 as a novel acid‐base transporter in ADPKD. NHA2 expression was closely associated with well‐established ADPKD mechanisms and signalling pathways (e.g.…”

“…In the current issue of The Journal of Physiology , Prasad et al . () have further explored the involvement of acid‐base transporters in PKD. The authors report that the human NHA2 orthologue of the bacterial NhaA (NHE superfamily) is upregulated in human ADPKD, and NHA2 stimulates cyst growth in an in vitro cell model.…”

“…Using bioinformatics and an impartially designed approach, Prasad et al . () identified pharmacological agents such as caffeine and theophylline (methyl xanthines) that stimulated NHA2 gene expression, as well as dexamethasone and valproic acid that inhibited expression. These agents all influence intracellular Ca 2+ in agreement with the 3D‐culture model data.…”

“…Prasad et al . () have reported exciting new results that allude to multifaceted roles of acid‐base transporters in PKD. In addition to moving solutes directly and contributing to reabsorption/secretion, these transporters also regulate intracellular pH, which is well known to influence cell function, signalling pathways, and proliferation – all of which are key elements of PKD.…”