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

Apical mislocation of the ubiquitous transport enzyme Na,K-ATPase has been implicated as a feature of cyst development in in vitro studies of human polycystic kidney disease (PKD) epithelia. We undertook an immunohistochemical study of murine glucocorticoid-induced PKD, the pcy mouse, the cpk mouse, and the diphenylthiazole (DPT)-induced rat models of PKD to determine if this feature was common to these models of cyst development. Distribution of Na,K-ATPase was determined with a polyclonal anti-Na,K-ATPase antibody and a nickel-silver-enhanced peroxidase color development system. Results were documented objectively with densitometric techniques. Control animals appropriate to the age, strain, and species of the experimental groups demonstrated the expected polar distribution of Na,K-ATPase to the basolateral surface. This distribution was more marked in mature animals. Tubular dilatation and cystic change, however, were associated with increased apical Na,K-ATPase in all models. The murine models demonstrated decreased basolateral staining for Na,K-ATPase compared with controls, although this was not a feature of the DPT rat model. Abnormal location of Na,K-ATPase is a shared feature of a variety of animal models and human PKD. This may contribute to abnormal fluid and electrolyte flux favoring cyst formation or may represent expression of a less differentiated renal tubule epithelial phenotype.

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FP058AN OLIVE LEAF EXTRACT REDUCES INFLAMMATION AND FIBROSIS IN AN IN VITRO MODEL OF AUTOSOMAL DOMINANT POLYCYSTIC KIDNEY DISEASE

Nephrology Dialysis Transplantation ◽

10.1093/ndt/gfz106.fp058 ◽

2019 ◽

Vol 34 (Supplement_1) ◽

Author(s):

Giuseppina Toteda ◽

Anna Perri ◽

Simona Lupinacci ◽

Donatella Vizza ◽

Antonella La Russa ◽

...

Keyword(s):

Kidney Disease ◽

Leaf Extract ◽

In Vitro Model ◽

Autosomal Dominant ◽

Olive Leaf ◽

Polycystic Kidney ◽

Olive Leaf Extract ◽

Autosomal Dominant Polycystic Kidney ◽

Vitro Model

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

10.1101/364679 ◽

2018 ◽

Author(s):

Hari Prasad ◽

Donna K. Dang ◽

Kalyan C. Kondapalli ◽

Niranjana Natarajan ◽

Valeriu Cebotaru ◽

...

Keyword(s):

Kidney Disease ◽

Polycystic Kidney Disease ◽

Mdck Cells ◽

Ectopic Expression ◽

Fluid Secretion ◽

Dominant Negative ◽

Polycystic Kidney ◽

Cyst Size ◽

Cyst Development

SummaryAutosomal 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 show 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 a dominant negative, membrane-anchored C-terminal tail fragment of PC1 elevated NHA2. We show that NHA2 is a target of Ca2+/NFAT signaling and is transcriptionally induced by methylxanthine drugs such as caffeine and theophylline, which are contraindicated in ADPKD patients. Finally, we observe 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 PKD and reveal a potential new regulator of transepithelial salt and water transport in the kidney.

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An inhibitor of histone deacetylase 6 activity, ACY-1215, reduces cAMP and cyst growth in polycystic kidney disease

AJP Renal Physiology ◽

10.1152/ajprenal.00186.2017 ◽

2017 ◽

Vol 313 (4) ◽

pp. F997-F1004 ◽

Cited By ~ 11

Author(s):

Murali K. Yanda ◽

Qiangni Liu ◽

Liudmila Cebotaru

Keyword(s):

Kidney Disease ◽

Histone Deacetylase ◽

Polycystic Kidney Disease ◽

Cyst Formation ◽

Histone Deacetylase 6 ◽

Polycystic Kidney ◽

Autosomal Dominant Polycystic Kidney ◽

Vitro Model ◽

Cyst Growth

Adult-onset autosomal-dominant polycystic kidney disease (ADPKD) is caused by mutations in either the PKD1 or PKD2 gene, leading to malfunction of their gene products, polycystin 1 or 2. Histone deacetylase 6 (HDAC6) expression and activity are increased in PKD1 mutant renal epithelial cells. Here we studied the effect of ACY-1215, a specific HDAC6 inhibitor, on cyst growth in ADPKD. Treatment with ACY-1215 slowed cyst growth in a mouse model of ADPKD that forms massive cysts within 3 wk after knockout of polycystin 1 function. It also prevented cyst formation in MDCK.2 cells, an in vitro model of cystogenesis, and in an ADPKD cell line derived from the proximal tubules from a pkd1−/−.mouse (PN cells). In PN cells ACY-1215 also reduced the size of already established cysts. We found that ACY-1215 lowered cAMP levels and protein expression of adenylyl cyclase 6. Our results suggest that HDAC6 could potentially serve as a therapeutic target in ADPKD.

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Autosomal dominant polycystic kidney disease—in vitro culture of cyst-lining epithelial cells

Virchows Archiv B Cell Pathology Including Molecular Pathology ◽

10.1007/bf02890421 ◽

1992 ◽

Vol 61 (1) ◽

pp. 189-199 ◽

Cited By ~ 3

Author(s):

R. Klingel ◽

S. Störkel ◽

W. Dippold ◽

H. -J. Rumpelt ◽

R. Moll ◽

...

Keyword(s):

Kidney Disease ◽

Epithelial Cells ◽

In Vitro Culture ◽

Polycystic Kidney Disease ◽

Autosomal Dominant ◽

Polycystic Kidney ◽

Autosomal Dominant Polycystic Kidney ◽

Cyst Lining

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Obacunone Retards Renal Cyst Development in Autosomal Dominant Polycystic Kidney Disease by Activating NRF2

Antioxidants ◽

10.3390/antiox11010038 ◽

2021 ◽

Vol 11 (1) ◽

pp. 38

Author(s):

Zhiwei Qiu ◽

Jinzhao He ◽

Guangying Shao ◽

Jiaqi Hu ◽

Xiaowei Li ◽

...

Keyword(s):

Kidney Disease ◽

Polycystic Kidney Disease ◽

Autosomal Dominant ◽

Renal Cyst ◽

Renal Diseases ◽

Therapeutic Drug ◽

Dependent Manner ◽

Polycystic Kidney ◽

Autosomal Dominant Polycystic Kidney ◽

Cyst Development

Autosomal dominant polycystic kidney disease (ADPKD) is a common inherited disease characterized by progressive enlargement of fluid-filled cysts derived from renal tubular epithelial cells, which has become the fourth leading cause of end-stage renal diseases. Currently, treatment options for ADPKD remain limited. The purpose of this study was to discover an effective therapeutic drug for ADPKD. With virtual screening, Madin-Darby canine kidney (MDCK) cyst model, embryonic kidney cyst model and kidney-specific Pkd1 knockout mouse (PKD) model, we identified obacunone as a candidate compound for ADPKD drug discovery from a natural antioxidant compound library. In vitro experiments showed that obacunone significantly inhibited cyst formation and expansion of MDCK cysts and embryonic kidney cysts in a dose-dependent manner. In vivo, obacunone treatment significantly reduced the renal cyst development in PKD mice. Western blot and morphological analysis revealed that obacunone served as a NRF2 activator in ADPKD, which suppressed lipid peroxidation by up-regulating GPX4 and finally restrained excessive cell proliferation by down-regulating mTOR and MAPK signaling pathways. Experimental data demonstrated obacunone as an effective renal cyst inhibitor for ADPKD, indicating that obacunone might be developed into a therapeutic drug for ADPKD treatment.

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Discovery and preclinical evaluation of anti-miR-17 oligonucleotide RGLS4326 for the treatment of polycystic kidney disease

Nature Communications ◽

10.1038/s41467-019-11918-y ◽

2019 ◽

Vol 10 (1) ◽

Cited By ~ 19

Author(s):

Edmund C. Lee ◽

Tania Valencia ◽

Charles Allerson ◽

Annelie Schairer ◽

Andrea Flaten ◽

...

Keyword(s):

Kidney Disease ◽

Polycystic Kidney Disease ◽

Collecting Duct ◽

Treatment Options ◽

Subcutaneous Administration ◽

Polycystic Kidney ◽

Short Oligonucleotide ◽

Mrna Targets ◽

Stage Renal Disease

Abstract Autosomal dominant polycystic kidney disease (ADPKD), caused by mutations in either PKD1 or PKD2 genes, is one of the most common human monogenetic disorders and the leading genetic cause of end-stage renal disease. Unfortunately, treatment options for ADPKD are limited. Here we report the discovery and characterization of RGLS4326, a first-in-class, short oligonucleotide inhibitor of microRNA-17 (miR-17), as a potential treatment for ADPKD. RGLS4326 is discovered by screening a chemically diverse and rationally designed library of anti-miR-17 oligonucleotides for optimal pharmaceutical properties. RGLS4326 preferentially distributes to kidney and collecting duct-derived cysts, displaces miR-17 from translationally active polysomes, and de-represses multiple miR-17 mRNA targets including Pkd1 and Pkd2. Importantly, RGLS4326 demonstrates a favorable preclinical safety profile and attenuates cyst growth in human in vitro ADPKD models and multiple PKD mouse models after subcutaneous administration. The preclinical characteristics of RGLS4326 support its clinical development as a disease-modifying treatment for ADPKD.

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