epidermal-growth-factor and Polycystic-Kidney--Autosomal-Dominant

Members of the epidermal growth factor (EGF) family bind to ErbB (EGFR) family receptors which play an important role in the regulation of various fundamental cell processes including cell proliferation and differentiation. The normal rodent kidney has been shown to express at least three members of the ErbB receptor family and is a major site of EGF ligand synthesis. Polycystic kidney disease (PKD) is a group of diseases caused by mutations in single genes and is characterized by enlarged kidneys due to the formation of multiple cysts in both kidneys. Tubule cells proliferate, causing segmental dilation, in association with the abnormal deposition of several proteins. One of the first abnormalities described in cell biological studies of PKD pathogenesis was the abnormal mislocalization of the EGFR in cyst lining epithelial cells. The kidney collecting duct (CD) is predominantly an absorptive epithelium where electrogenic Na(+) entry is mediated by the epithelial Na(+) channel (ENaC). ENaC-mediated sodium absorption represents an important ion transport pathway in the CD that might be involved in the development of PKD. A role for EGF in the regulation of ENaC-mediated sodium absorption has been proposed. However, several investigations have reported contradictory results indicating opposite effects of EGF and its related factors on ENaC activity and sodium transport. Recent advances in understanding how proteins in the EGF family regulate the proliferation and sodium transport in normal and PKD epithelial cells are discussed here. This article is part of a Special Issue entitled: Polycystic Kidney Disease.

Topics: Animals; Cell Proliferation; Epidermal Growth Factor; Epithelial Cells; Epithelial Sodium Channels; ErbB Receptors; Humans; Ion Transport; Kidney; Models, Biological; Polycystic Kidney Diseases; Polycystic Kidney, Autosomal Dominant; Polycystic Kidney, Autosomal Recessive; Sodium; TRPP Cation Channels

In this study, we report a model of spontaneous cyst formation in vitro and a procedure to obtain large quantities of cysts from polycystic rat kidney cells. Furthermore, we assess the effects of epidermal growth factor, a modulator of morphogenesis, and of taxol, a stabilizer of microtubules, which has recently been proposed as a useful treatment of human polycystic kidney disease (PKD). It is anticipated that data generated from in vitro studies using cysts from PKD-affected rat kidneys may yield further insights to the pathophysiological and cellular basis of fatal renal cyst formation processes, and may lead to specific therapeutic strategies directed at controlling the growth of cysts, thereby reducing the number of animal tests.

Topics: Animals; Antineoplastic Agents, Phytogenic; Cell Aggregation; Cells, Cultured; Epidermal Growth Factor; Kidney; Male; Paclitaxel; Polycystic Kidney, Autosomal Dominant; Rats; Rats, Mutant Strains; Time Factors

Age- and height-adjusted total kidney volume is currently considered the best prognosticator in patients with autosomal dominant polycystic kidney disease. We tested the ratio of urinary epidermal growth factor and monocyte chemotactic peptide 1 for the prediction of the Mayo Clinic Imaging Classes.. Urinary epidermal growth factor and monocyte chemotactic peptide 1 levels were measured in two independent cohorts (discovery, n = 74 and validation set, n = 177) and healthy controls (n = 59) by immunological assay. Magnetic resonance imaging parameters were used for total kidney volume calculation and the Mayo Clinic Imaging Classification defined slow (1A-1B) and fast progressors (1C-1E). Microarray and quantitative gene expression analysis were used to test epidermal growth factor and monocyte chemotactic peptide 1 gene expression.. Baseline ratio of urinary epidermal growth factor and monocyte chemotactic peptide 1 correlated with total kidney volume adjusted for height (r = - 0.6, p < 0.001), estimated glomerular filtration rate (r = 0.69 p < 0.001), discriminated between Mayo Clinic Imaging Classes (p < 0.001), and predicted the variation of estimated glomerular filtration rate at 10 years (r = - 0.51, p < 0.001). Conditional Inference Trees identified cut-off levels of the ratio of urinary epidermal growth factor and monocyte chemotactic peptide 1 for slow and fast progressors at > 132 (100% slow) and < 25.76 (89% and 86% fast, according to age), with 94% sensitivity and 66% specificity (p = 6.51E-16). Further, the ratio of urinary epidermal growth factor and monocyte chemotactic peptide 1 at baseline showed a positive correlation (p = 0.006, r = 0.36) with renal outcome (delta-estimated glomerular filtration rate per year, over a mean follow-up of 4.2 ± 1.2 years). Changes in the urinary epidermal growth factor and monocyte chemotactic peptide 1 were mirrored by gene expression levels in both human kidney cysts (epidermal growth factor: - 5.6-fold, fdr = 0.001; monocyte chemotactic peptide 1: 3.1-fold, fdr = 0.03) and Pkd1 knock-out mouse kidney (Egf: - 14.8-fold, fdr = 2.37E-20, Mcp1: 2.8-fold, fdr = 6.82E-15).. The ratio of urinary epidermal growth factor and monocyte chemotactic peptide 1 is a non-invasive pathophysiological biomarker that can be used for clinical risk stratification in autosomal dominant polycystic kidney disease.

Topics: Animals; Disease Progression; Epidermal Growth Factor; Humans; Kidney; Mice; Monocytes; Polycystic Kidney, Autosomal Dominant

The epidermal growth factor receptor (EGFR) pathway is involved in kidney tissue repair and growth. Preclinical interventional data and scarce human data have suggested a role for this pathway in the pathophysiology of autosomal dominant polycystic kidney disease (ADPKD), while other data have suggested that its activation is causally linked to repair of damaged kidney tissue. We hypothesize that urinary EGFR ligands, as a reflection of EGFR activity, are associated with kidney function decline in ADPKD in the context of tissue repair following injury, and as the disease progresses as a sign of insufficient repair.. In the present study, we measured the EGFR ligands, EGF and heparin binding-EGF (HB-EGF), in 24-h urine samples of 301 ADPKD patients and 72 age- and sex-matched living kidney donors to dissect the role of the EGFR pathway in ADPKD. During a median follow-up of 2.5 years, the association of urinary EGFR ligand excretion with annual change in estimated glomerular filtration rate (eGFR) and height-adjusted total kidney volume in ADPKD patients was analyzed using mixed-models methods, and the expression of three closely related EGFR family receptors in ADPKD kidney tissue was investigated by immunohistochemistry. Additionally, the effect of reducing renal mass (after kidney donation), was assessed to investigate whether urinary EGF matches this reduction and thus reflects the amount of remaining healthy kidney tissue.. At baseline, urinary HB-EGF did not differ between ADPKD patients and healthy controls (P = .6), whereas a lower urinary EGF excretion was observed in ADPKD patients [18.6 (11.8-27.8)] compared with healthy controls [51.0 (34.9-65.4) μg/24 h, P < .001]. Urinary EGF was positively associated with baseline eGFR (R = 0.54, P < .001) and a lower EGF was strongly associated with a more rapid GFR decline, even when adjusted for ADPKD severity markers (β = 1.96, P < .001), whereas HB-EGF was not. Expression of the EGFR, but not other EGFR-related receptors, was observed in renal cysts but was absent in non-ADPKD kidney tissue. Finally, unilateral nephrectomy resulted in a decrease of 46.4 (-63.3 to -17.6) % in urinary EGF excretion, alongside a decrease of 35.2 ± 7.2% in eGFR and 36.8 ± 6.9% in measured GFR (mGFR), whereas maximal mGFR (measured after dopamine induced hyperperfusion) decreased by 46.1 ± 7.8% (all P < .001).. Our data suggest that lower urinary EGF excretion may be a valuable novel predictor for kidney function decline in patients with ADPKD.

Topics: Disease Progression; Epidermal Growth Factor; Glomerular Filtration Rate; Heparin-binding EGF-like Growth Factor; Humans; Kidney; Patient Acuity; Polycystic Kidney, Autosomal Dominant

Autosomal dominant polycystic kidney disease (ADPKD) is the most common hereditary kidney disorder leading to deterioration of kidney function and end stage kidney disease (ESKD). A number of molecular processes are dysregulated in ADPKD but the exact mechanism of disease progression is not fully understood. We measured protein biomarkers being linked to ADPKD-associated molecular processes via ELISA in urine and serum in a cohort of ADPKD patients as well as age, gender and eGFR matched CKD patients and healthy controls. ANOVA and

Topics: Adult; Aged; Aged, 80 and over; Angiotensinogen; Apelin; Arginine Vasopressin; Biomarkers; Epidermal Growth Factor; Female; Humans; Male; Middle Aged; Polycystic Kidney, Autosomal Dominant; Vascular Endothelial Growth Factor A

Topics: Adult; Aged; Arabs; Calcium Channels; Case-Control Studies; Cystic Fibrosis Transmembrane Conductance Regulator; DNA Mutational Analysis; Epidermal Growth Factor; Exome Sequencing; Exons; Female; Humans; Male; Middle Aged; Mutation, Missense; Polycystic Kidney, Autosomal Dominant; Receptors, Cell Surface; Saudi Arabia; Tomography, X-Ray Computed; TRPP Cation Channels; Tuberous Sclerosis Complex 2 Protein

Recent animal experiments suggest that dysregulation of the EGF receptor pathway plays a role in the pathophysiology of autosomal dominant polycystic kidney disease (ADPKD). Research on EGF receptor ligands in humans with ADPKD is lacking. EGF receptor ligands were measured in patients with ADPKD at baseline and after treatment with a vasopressin V2 receptor antagonist (V2RA) because this information might provide a rationale for future V2RA combination therapy.. Blood and urine concentrations of the EGF receptor ligands heparin-binding (HB)-EGF, EGF, and TGF-α were measured by ELISAs in 27 patients with ADPKD who participated in a single-center study investigating a V2RA in 2011-2013 and in 27 controls who were selected from a general population-based observational study. Cyst fluid concentrations were also measured. In patients with ADPKD, ligands were measured at baseline, after 3-week treatment with a V2RA, and 3 weeks after drug withdrawal. The measured GFR (mGFR) was determined by iothalamate infusion, and total kidney volume was measured by magnetic resonance imaging.. Urinary HB-EGF excretion and plasma concentration were higher in patients with ADPKD than in controls (median, 1.4 [interquartile range, 1.2-1.9] versus 0.6 [0.4-0.8] µg/24 hours [P<0.001] and 157.9 [83.1-225.9] versus 77.2 [37.2-174.3] pg/ml [P=0.04]). In contrast, urinary EGF excretion and plasma EGF concentration were lower in patients with ADPKD, whereas TGF-α did not differ between patients and controls. Higher HB-EGF excretion was correlated with more severe disease, assessed as lower mGFR (r=-0.39; P=0.05), higher total kidney volume (r=0.39; P=0.05), and higher urinary excretion of albumin and heart-type fatty acid-binding protein, whereas higher EGF excretion and TGF-α excretion were negatively correlated with disease severity. During V2RA treatment, HB-EGF excretion increased (from 1.4 [1.2-1.9] to 2.4 [2.1-3.1] µg/24 hours; P<0.001).. In patients with ADPKD, higher urinary HB-EGF excretion is correlated with more severe disease. Whether this association is causal needs to be investigated in intervention studies.

Topics: Adult; Aged; Antidiuretic Hormone Receptor Antagonists; Benzazepines; Case-Control Studies; EGF Family of Proteins; Epidermal Growth Factor; Female; Glomerular Filtration Rate; Heparin-binding EGF-like Growth Factor; Humans; Kidney; Magnetic Resonance Imaging; Male; Middle Aged; Organ Size; Polycystic Kidney, Autosomal Dominant; Severity of Illness Index; Tolvaptan; Transforming Growth Factor alpha

In autosomal dominant polycystic kidney disease (ADPKD), aberrant proliferation of the renal epithelial cells is responsible for the formation of numerable fluid-filled cysts, massively enlarged kidneys, and progressive loss of renal function. cAMP agonists, including arginine vasopressin, accelerate cyst epithelial cell proliferation through protein kinase A activation of the B-Raf/MEK/extracellular signal-regulated kinase (ERK) signaling pathway. The mitogenic effect of cAMP is equally potent and additive to growth factor stimulation. Here, we determined whether Sorafenib (BAY 43-9006), a small molecule Raf inhibitor, inhibits proliferation of cells derived from the cysts of human ADPKD kidneys. We found that nanomolar concentrations of Sorafenib reduced the basal activity of ERK, inhibited cAMP-dependent activation of B-Raf and MEK/ERK signaling, and caused a concentration-dependent inhibition of cell proliferation induced by cAMP, epidermal growth factor, or the combination of the two agonists. Sorafenib completely blocked in vitro cyst growth of human ADPKD cystic cells cultured within a three-dimensional collagen gel. These data demonstrate that cAMP-dependent proliferation of human ADPKD cyst epithelial cells is blocked by Sorafenib and suggest that small molecule B-Raf inhibitors may be a therapeutic option to reduce the mitogenic effects of cAMP on cyst expansion.

Topics: Benzenesulfonates; Blotting, Western; Cell Proliferation; Cells, Cultured; Cyclic AMP-Dependent Protein Kinases; Enzyme Activation; Epidermal Growth Factor; Epithelial Cells; Humans; Mitogen-Activated Protein Kinases; Niacinamide; Phenylurea Compounds; Polycystic Kidney, Autosomal Dominant; Protein Kinase Inhibitors; Proto-Oncogene Proteins B-raf; Proto-Oncogene Proteins c-raf; Pyridines; Signal Transduction; Sorafenib

Progressive renal enlargement due to the growth of innumerable fluid-filled cysts is a central pathophysiological feature of autosomal dominant polycystic kidney disease (ADPKD). These epithelial neoplasms enlarge slowly and damage noncystic parenchyma by mechanisms that have not been clearly defined. In a microarray analysis of cultured human ADPKD cyst epithelial cells, periostin mRNA was overexpressed 15-fold compared with normal human kidney (NHK) cells. Periostin, initially identified in osteoblasts, is not expressed in normal adult kidneys but is expressed transiently during renal development. We found periostin in cyst-lining cells in situ in the extracellular matrix adjacent to the cysts and within cyst fluid. ADPKD cells secreted periostin across luminal and basolateral plasma membranes. Periostin increased proliferation of cyst epithelial cells 27.9 +/- 3.1% (P < 0.001) above baseline and augmented in vitro cyst growth but did not affect proliferation of normal renal cells. Expression of alphaV-integrin, a periostin receptor, was ninefold higher in ADPKD cells compared with NHK cells, and antibodies that block alphaV-integrin inhibited periostin-induced cell proliferation. We conclude that periostin is a novel autocrine mitogen secreted by mural epithelial cells with the potential to accelerate cyst growth and promote interstitial remodeling in ADPKD.

Topics: Antibodies; Cell Adhesion Molecules; Cell Cycle; Cell Proliferation; Cells, Cultured; Colforsin; Cyclic AMP; Cyst Fluid; Cysts; Epidermal Growth Factor; Epithelial Cells; Extracellular Matrix; Flow Cytometry; Gene Expression; Humans; Immunohistochemistry; Integrin alphaV; Kidney; Polycystic Kidney, Autosomal Dominant; Recombinant Proteins; Reverse Transcriptase Polymerase Chain Reaction; Transforming Growth Factor beta

Autosomal dominant polycystic kidney disease (ADPKD) is characterized by the formation of multiple fluid-filled cysts that expand over time and destroy the renal architecture. Loss or mutation of polycystin-1 or polycystin-2, the respective proteins encoded by the ADPKD genes PKD1 and PKD2, is associated with most cases of ADPKD. Thus, the polycystin proteins likely play a role in cell proliferation and morphogenesis. Recent studies indicate that polycystin-1 is involved in these processes, but little is known about the role played by polycystin-2. To address this question, we created a number of related cell lines variable in their expression of polycystin-2. We show that the basal and epidermal growth factor-stimulated rate of cell proliferation is higher in cells that do not express polycystin-2 versus those that do, indicating that polycystin-2 acts as a negative regulator of cell growth. In addition, cells not expressing polycystin-2 exhibit significantly more branching morphogenesis and multicellular tubule formation under basal and hepatocyte growth factor-stimulated conditions than their polycystin-2-expressing counterparts, suggesting that polycystin-2 may also play an important role in the regulation of tubulogenesis. Cells expressing a channel mutant of polycystin-2 proliferated faster than those expressing the wild-type protein, but exhibited blunted tubule formation. Thus, the channel activity of polycystin-2 may be an important component of its regulatory machinery. Finally, we show that polycystin-2 regulation of cell proliferation appears to be dependent on its ability to prevent phosphorylated extracellular-related kinase from entering the nucleus. Our results indicate that polycystin-2 is necessary for the proper growth and differentiation of kidney epithelial cells and suggest a possible mechanism for the cyst formation seen in ADPKD2.

Topics: Cell Differentiation; Cell Division; Cell Line; Cysts; Epidermal Growth Factor; Epithelial Cells; Extracellular Signal-Regulated MAP Kinases; Gene Expression; Hepatocyte Growth Factor; Humans; Kidney Tubules; Membrane Proteins; Polycystic Kidney, Autosomal Dominant; TRPP Cation Channels

The proliferation of mural epithelial cells is a major cause of progressive cyst enlargement in autosomal-dominant polycystic kidney disease (ADPKD). Adenosine 3', 5' cyclic monophosphate (cAMP) stimulates the proliferation of cells from ADPKD cysts, but not cells from normal human kidney cortex (HKC), through the activation of protein kinase A (PKA), mitogen-activated protein kinase kinase (MEK), and extracellular signal-regulated kinase (ERK/MAPK). In the current study, we examined the signaling pathway between PKA and MEK in ADPKD and HKC cells.. Primary cultures of human ADPKD and HKC cells were prepared from nephrectomy specimens. We determined the effects of cAMP and epidermal growth factor (EGF) on the activation of ERK, B-Raf and Raf-1 in ADPKD and HKC cells by immune kinase assay and Western blot.. 8-Br-cAMP increased phosphorylated ERK (2.7- +/- 0.6-fold, N = 7), and B-Raf kinase activity (3.6- +/- 1.1-fold, N = 5) in cells from ADPKD kidneys; levels of phosphorylated Raf-1 were not changed. Inhibition of PKA by H89 strikingly decreased cAMP-stimulated phosphorylation of ERK and B-Raf, and MAPK inhibition by PD98059 blocked the effect of the nucleotide to activate ERK. By contrast, in HKC cells 8-Br-cAMP did not activate B-Raf and ERK. EGF stimulated the phosphorylation of ERK and Raf-1 in both ADPKD and HKC cells, but had no effect on B-Raf. 8-Br-cAMP and EGF conjointly increased ERK activation above that of either agonist alone in ADPKD cells, and this combined effect was abolished by PD98059, indicating that ERK was activated by EGF- and cAMP-responsive cascades that converge at MAPK.. cAMP activates ERK and increases proliferation of ADPKD epithelial cells, but not cells from normal human kidney cortex, through the sequential phosphorylation of PKA, B-Raf and MAPK in a pathway separate from, but complementary to, the classical receptor tyrosine kinase cascade. Consequently, cAMP and EGF have great potential to accelerate the progressive enlargement of renal cysts.

Topics: 8-Bromo Cyclic Adenosine Monophosphate; Cells, Cultured; Cyclic AMP; Drug Interactions; Epidermal Growth Factor; Epithelial Cells; Female; Humans; Kidney Cortex; Male; MAP Kinase Signaling System; Middle Aged; Mitogen-Activated Protein Kinases; Polycystic Kidney, Autosomal Dominant; Proto-Oncogene Proteins B-raf; Proto-Oncogene Proteins c-raf

Autosomal-dominant medullary cystic kidney disease type 2 (MCKD2) is a tubulointerstitial nephropathy that causes renal salt wasting, hyperuricemia, gout, and end-stage renal failure in the fifth decade of life. The chromosomal locus for MCKD2 was localized on chromosome 16p12. Within this chromosomal region, Uromodulin (UMOD) was located as a candidate gene. UMOD encodes the Tamm-Horsfall protein. By sequence analysis, one group formerly excluded UMOD as the disease-causing gene. In contrast, recently, another group described mutations in the UMOD gene as responsible for MCKD2 and familial juvenile hyperuricemic nephropathy (FJHN).. Haplotype analysis for linkage to MCKD2 was performed in 25 MCKD families. In the kindreds showing linkage to the MCKD2 locus on chromosome 16p12, mutational analysis of the UMOD gene was performed by exon polymerase chain reaction (PCR) and direct sequencing.. In 19 families, haplotype analysis was compatible with linkage to the MCKD2 locus. All these kindreds were examined for mutations in the UMOD gene. In three different families, three novel heterozygous mutations in the UMOD gene were found and segregated with the phenotype in affected individuals. Mutations were found only in exon 4.. We confirm the UMOD gene as the disease-causing gene for MCKD2. All three novel mutations were found in the fourth exon of UMOD, in which all mutations except one (this is located in the neighboring exon 5) published so far are located. These data point to a specific role of exon 4 encoded sequence of UMOD in the generation of the MCKD2 renal phenotype.

Topics: Adolescent; Amino Acid Sequence; Base Sequence; Child; Child, Preschool; Epidermal Growth Factor; Exons; Female; Genetic Linkage; Haplotypes; Humans; Male; Molecular Sequence Data; Mucoproteins; Multigene Family; Pedigree; Phenotype; Polycystic Kidney, Autosomal Dominant; Protein Structure, Tertiary; Uromodulin

Epithelial cells cultured from the renal cysts of patients with autosomal dominant polycystic kidney disease (ADPKD) secrete fluid via a process stimulated by adenosine 3',5'-cyclic monophosphate (cAMP). We have investigated the hypothesis that fluid secretion by these cells is dependent on cAMP-mediated chloride secretion. Individual cultured ADPKD cells were suspended within a polymerized collagen matrix and stimulated to form cysts. Individual cultured cysts were placed in a chamber on the stage of an inverted microscope equipped with epifluorescent and video analysis attachments. The rate of fluid secretion, cell volume and changes in intracellular Cl- were measured. In the absence of secretagogues, fluid was absorbed from the cyst cavity (-2.36 +/- 0.64 nl/min/cm2 inner surface area). 8-Bromoadenosine 3',5'-cyclic monophosphate (8-Br-cAMP) plus 3-isobutyl-1-methlyxanthine (IBMX) induced a rapid reversal in the net movement of fluid to secretion (6.79 +/- 1.28 nl/min/cm2). Bumetanide reversibly reduced fluid secretion to 0.95 +/- 0.60 nl/min/cm2. Cell volume rapidly decreased by 7.5 +/- 0.9% with the initiation of secretion and bumetanide caused an additional loss (4.2 +/- 1.0%). Furosemide had a similar effect on forskolin-induced fluid secretion. Cellular chloride concentration was monitored with the use of the indicator, 6-methoxy-N-ethylquinolinium chloride (MEQ). Removal of Cl- from the bath reduced intracellular [Cl-] (MEQ fluorescence increased by 11.4 +/- 2.3%). In cysts pretreated with furosemide to prevent Cl- entry, the application of forskolin caused a decrease in Cl- concentration (MEQ fluorescence increased by 9.3 +/- 2.6%). Using monolayers of cultured ADPKD cells, grown on permeant supports, we compared the changes in short circuit current (ISC) induced by forskolin in the presence and absence of external Cl-. Forskolin increased ISC (from 8.9 +/- 2.7 to 10.6 +/- 2.7 microA/cm2) in the presence of Cl-, but did not significantly affect ISC in its absence. These data indicate that cultured ADPKD cells can direct fluid transport in either the absorptive or the secretory direction, and that cAMP stimulates secretion and this secretion is accompanied by a net loss of cell solute. Inhibition of secretion by bumetanide or furosemide caused an additional loss of cell solute, including Cl-. The ionic transepithelial current induced by forskolin is dependent on the presence of Cl-. These data support the thesis that chloride secretion drives flu

Topics: Bumetanide; Cells, Cultured; Chlorides; Colforsin; Cysts; Electric Conductivity; Epidermal Growth Factor; Exudates and Transudates; Furosemide; Humans; Kidney; Polycystic Kidney, Autosomal Dominant

The underlying mechanism of the hyperproliferative response of human autosomal dominant polycystic kidney disease (ADPKD) epithelia was studied. Epidermal growth factor (EGF) protein is highly expressed in ADPKD cyst epithelia in vivo, and primary cultures are hyperesponsive to mitogenic stimulation by EGF in vitro. Doses of > 1 ng/ml EGF were highly mitogenic to ADPKD epithelia. 3H-labeled thymidine proliferation assays showed that cyst fluids and ADPKD epithelial cell-conditioned media also stimulated renal epithelial cell proliferation and contained EGF immunoreactivity (6, 30, and 37 kDa) as detected by Western blots. Radioimmunoassays detected mean levels of 2.87 and 1.4 ng/ml EGF in cyst fluids from early (proliferative) and end-stage ADPKD cysts, respectively. Scatchard analysis of 125I-labeled EGF binding to apical and basolateral membrane showed high-affinity binding to basolateral membranes of normal and ADPKD kidneys but additional unique high-affinity receptor binding to apical membranes of ADPKD but not normal kidneys. Cross-linking analysis and antiphosphotyrosine Western analysis demonstrated functionally active apical EGF receptors at 150-170 kDa. These results suggest mediation of cyst expansion via an autocrine loop involving EGF synthesis and processing by cyst epithelial cells, apical secretion into cyst lumens, and subsequent binding to and phosphorylation of apical membrane EGF receptors. These findings are consistent with a membrane protein polarization defect in ADPKD cyst epithelia.

Topics: Body Fluids; Cell Division; Cell Membrane; Cell Polarity; Cells, Cultured; Culture Media, Conditioned; Epidermal Growth Factor; Epithelium; ErbB Receptors; Humans; Kidney; Polycystic Kidney, Autosomal Dominant; Radioimmunoassay; Tissue Distribution

Topics: Creatinine; Epidermal Growth Factor; Humans; Kidney Failure, Chronic; Polycystic Kidney, Autosomal Dominant

Topics: Animals; Biomarkers; Blotting, Northern; Clusterin; Disease Models, Animal; Electrophoresis, Agar Gel; Epidermal Growth Factor; Female; Glycoproteins; Kidney; Male; Molecular Chaperones; Nucleic Acid Hybridization; Polycystic Kidney, Autosomal Dominant; Rats; Rats, Inbred Strains; RNA; RNA, Messenger

We determined the concentrations of immunoreactive epidermal growth factor (EGF), endothelin (ET), and cyclic adenosine monophosphate (cAMP) in cyst fluid from 10 nondialyzed patients with autosomal dominant polycystic kidney disease (ADPKD). The concentrations of ET and cAMP in low-sodium cyst fluid (cyst sodium concentration < 100 mEq/L) were significantly increased compared with those in high-sodium cyst fluid (cyst sodium concentration > or = 100 mEq/L), but the concentration of EGF did not differ between the two. There was no relationship between cyst fluid volume and EGF, ET, cAMP values in ADPKD cases. Significant variations of cyst fluid concentrations of EGF, ET, and cAMP were observed among the patients. However, the differences of these variables between the right and left kidney of the individual subjects were unremarkable. Serum levels of creatinine and urea nitrogen showed significant negative correlations with EGF, but not with ET and cAMP. The present data demonstrate the differences in cytokine activity among cysts in ADPKD cases, suggesting the heterogeneity of cystogenesis along the tubular segments in this particular disease.

Topics: Adult; Blood Urea Nitrogen; Body Fluids; Creatinine; Cyclic AMP; Endothelins; Epidermal Growth Factor; Female; Humans; Male; Middle Aged; Polycystic Kidney, Autosomal Dominant

Genetic polycystic kidney disease (ADPKD) is an autosomal dominant trait in man, the phenotypic expression of which is characterized by enormous cystic enlargement of renal tubules. Since this is, in part, a result of aberrant epithelial cell proliferation, the nature of this defect has been characterized by comparison of growth factor action on defined epithelial primary monolayer cultures derived from individually microdissected normal human renal proximal and distal tubules and ADPKD cyst-lining epithelia. Mitogenic assays showed an increased sensitivity of ADPKD epithelia to stimulation by the combination of the endocrine factors hydrocortisone (dexamethasone) and insulin, and Northern analysis suggested increased levels of insulin receptor steady state mRNA. The most potent, single mitogen was epidermal growth factor (EGF), and hypersensitivity to EGF in ADPKD epithelia was correlated with increased mRNA levels for EGF receptor in ADPKD kidneys. The localization in vivo of EGF immunoreactivity in ADPKD cyst-lining epithelia and in (apical) cyst fluids and the demonstration of EGF-receptor immunostaining and specific [125I]EGF binding to apical cell surfaces suggested an autocrine mechanism of growth stimulation by EGF in ADPKD epithelia. Transforming growth factor beta was an inhibitor of normal renal tubule proliferation but was unable to completely inhibit EGF stimulation in ADPKD cultures. Platelet-derived growth factor (PDGF) immunoreactivity which was also seen in ADPKD cyst epithelia, was not mitogenic to ADPKD epithelia but did stimulate division in ADPKD fibroblasts in vitro. This suggested a paracrine regulation of the observed fibrosis in ADPKD.

Topics: Blotting, Northern; Cell Division; Cells, Cultured; Epidermal Growth Factor; Epithelial Cells; Fluorescent Antibody Technique; Growth Substances; Hormones; Humans; Iodine Radioisotopes; Mitogens; Peptides; Polycystic Kidney, Autosomal Dominant; Radioligand Assay; RNA, Messenger