Small-molecule CFTR inhibitors slow cyst growth in polycystic kidney disease.

Cyst expansion in polycystic kidney disease (PKD) involves progressive fluid accumulation, which is believed to require chloride transport by the cystic fibrosis transmembrane conductance regulator (CFTR) protein. Herein is reported that small-molecule CFTR inhibitors of the thiazolidinone and glycine hydrazide classes slow cyst expansion in in vitro and in vivo models of PKD. More than 30 CFTR inhibitor analogs were screened in an MDCK cell model, and near-complete suppression of cyst growth was found by tetrazolo-CFTR(inh)-172, a tetrazolo-derived thiazolidinone, and Ph-GlyH-101, a phenyl-derived glycine hydrazide, without an effect on cell proliferation. These compounds also inhibited cyst number and growth by >80% in an embryonic kidney cyst model involving 4-d organ culture of embryonic day 13.5 mouse kidneys in 8-Br-cAMP-containing medium. Subcutaneous delivery of tetrazolo-CFTR(inh)-172 and Ph-GlyH-101 to neonatal, kidney-specific PKD1 knockout mice produced stable, therapeutic inhibitor concentrations of >3 microM in urine and kidney tissue. Treatment of mice for up to 7 d remarkably slowed kidney enlargement and cyst expansion and preserved renal function. These results implicate CFTR in renal cyst growth and suggest that CFTR inhibitors may hold therapeutic potential to reduce cyst growth in PKD.

[1]  J. Grantham,et al.  The secretion of fluid by renal cysts from patients with autosomal dominant polycystic kidney disease. , 1993, The New England journal of medicine.

[2]  G. Germino,et al.  The Molecular Basis of Focal Cyst Formation in Human Autosomal Dominant Polycystic Kidney Disease Type I , 1996, Cell.

[3]  D. Wallace,et al.  Epithelial transport in polycystic kidney disease. , 1998, Physiological reviews.

[4]  A. Verkman,et al.  In vivo pharmacology and antidiarrheal efficacy of a thiazolidinone CFTR inhibitor in rodents. , 2005, Journal of pharmaceutical sciences.

[5]  E. Avner,et al.  Molecular and cellular pathophysiology of autosomal recessive polycystic kidney disease (ARPKD) , 2006, Cell and Tissue Research.

[6]  R. Kucherlapati,et al.  Somatic Inactivation of Pkd2 Results in Polycystic Kidney Disease , 1998, Cell.

[7]  P. Wilson,et al.  Polycystic kidney disease. , 2004, The New England journal of medicine.

[8]  M. Arnaout.  Molecular genetics and pathogenesis of autosomal dominant polycystic kidney disease. , 2001, Annual review of medicine.

[9]  H. Kanetake,et al.  Human simple renal cyst fluid contains a cyst formation‐promoting activity for Madin–Darby canine kidney cells cultured in collagen gel , 1996, European journal of clinical investigation.

[10]  J. Mcateer,et al.  An in vitro test of the cell stretch-proliferation hypothesis of renal cyst enlargement. , 1995, Journal of the American Society of Nephrology : JASN.

[11]  Moshe Talpaz,et al.  Dasatinib (BMS-354825) Tyrosine Kinase Inhibitor Suppresses Invasion and Induces Cell Cycle Arrest and Apoptosis of Head and Neck Squamous Cell Carcinoma and Non–Small Cell Lung Cancer Cells , 2005, Clinical Cancer Research.

[12]  L. Galietta,et al.  Lectin conjugates as potent, nonabsorbable CFTR inhibitors for reducing intestinal fluid secretion in cholera. , 2007, Gastroenterology.

[13]  P. Harris,et al.  Molecular genetics of autosomal recessive polycystic kidney disease. , 2004, Molecular genetics and metabolism.

[14]  P. Gabow.  Autosomal dominant polycystic kidney disease. , 2010, The New England journal of medicine.

[15]  A. Chapman.  Autosomal dominant polycystic kidney disease: time for a change? , 2007, Journal of the American Society of Nephrology : JASN.

[16]  J. Grantham,et al.  Renal epithelial cyst formation and enlargement in vitro: dependence on cAMP. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[17]  J. Glockner,et al.  Autosomal dominant polycystic kidney disease coexisting with cystic fibrosis. , 2006, Journal of nephrology.

[18]  P. George-Hyslop,et al.  Mutations of PKD1 in ADPKD2 cysts suggest a pathogenic effect of trans-heterozygous mutations , 2000, Nature Genetics.

[19]  P. Igarashi,et al.  Genetics and pathogenesis of polycystic kidney disease. , 2002, Journal of the American Society of Nephrology : JASN.

[20]  E. Avner,et al.  New insights into the molecular pathophysiology of polycystic kidney disease. , 1999, Kidney international.

[21]  I. Gupta,et al.  Morphogenesis during mouse embryonic kidney explant culture. , 2003, Kidney international.

[22]  P. Igarashi,et al.  Epithelial-specific Cre/lox recombination in the developing kidney and genitourinary tract. , 2002, Journal of the American Society of Nephrology : JASN.

[23]  A. Verkman,et al.  Luminally active, nonabsorbable CFTR inhibitors as potential therapy to reduce intestinal fluid loss in cholera , 2006, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[24]  David N Sheppard,et al.  The relationship between cell proliferation, Cl- secretion, and renal cyst growth: a study using CFTR inhibitors. , 2004, Kidney international.

[25]  S. Somlo,et al.  Effective treatment of an orthologous model of autosomal dominant polycystic kidney disease , 2004, Nature Medicine.

[26]  S. Thibodeau,et al.  Cystic fibrosis and the phenotypic expression of autosomal dominant polycystic kidney disease. , 1998, American journal of kidney diseases : the official journal of the National Kidney Foundation.

[27]  A. Evan,et al.  Polarized epithelial cysts in vitro: a review of cell and explant culture systems that exhibit epithelial cyst formation. , 1988, Scanning microscopy.

[28]  B. Magenheimer,et al.  Early embryonic renal tubules of wild-type and polycystic kidney disease kidneys respond to cAMP stimulation with cystic fibrosis transmembrane conductance regulator/Na(+),K(+),2Cl(-) Co-transporter-dependent cystic dilation. , 2006, Journal of the American Society of Nephrology : JASN.

[29]  L. Cantley,et al.  Cyst formation and activation of the extracellular regulated kinase pathway after kidney specific inactivation of Pkd1. , 2008, Human molecular genetics.

[30]  A. Taddei,et al.  Discovery of Glycine Hydrazide Pore-occluding CFTR Inhibitors , 2004, The Journal of general physiology.

[31]  J. Riordan,et al.  Functional expression and apical localization of the cystic fibrosis transmembrane conductance regulator in MDCK I cells. , 1997, The Biochemical journal.

[32]  A. Taddei,et al.  Altered channel gating mechanism for CFTR inhibition by a high‐affinity thiazolidinone blocker , 2004, FEBS letters.

[33]  L. Rome,et al.  The cystic fibrosis transmembrane conductance regulator mediates transepithelial fluid secretion by human autosomal dominant polycystic kidney disease epithelium in vitro. , 1996, Kidney international.

[34]  J. Calvet,et al.  Renal epithelial fluid secretion and cyst growth: the role of cyclic AMP , 1989, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[35]  J. Grantham,et al.  Immunolocalization of ion transport proteins in human autosomal dominant polycystic kidney epithelial cells. , 1996, Proceedings of the National Academy of Sciences of the United States of America.