CYCLIC POLYPEPTIDE FOR THE TREATMENT OF PHA TYPE 1B

Abstract

A cyclic polypeptide comprising at least six contiguous amino acids from the amino acid sequence SEQ ID NO:1 Gly-Gln-Arg-Glu-Thr-Pro-Glu-Gly-Ala-Glu-Ala-Lys-Pro-Trp-Tyr for the treatment of autosomal recessive pseudohypoaldosteronism type 1 (PHA type1B) or for the restoration of the Na transport capacity of mutated loss-of-function ENaC.

Claims

1. A method of treating a disease, comprising: administering to a subject in need of treatment an effective amount of a cyclic polypeptide comprising at least six contiguous amino acids from the amino acid sequence SEQ ID NO:1
Gly-Gln-Arg-Glu-Thr-Pro-Glu-Gly-Ala-Glu-Ala-Lys-Pro-Trp-Tyr, the cyclic polypeptide treating autosomal recessive pseudohypoaldosteronism type 1 (PHA type1B) in the subject, or the cyclic polypeptide restoring the Na.sup.+ ion transport capacity of mutated loss-of-function ENaC in the subject.

2. The method according to claim 1, wherein the cyclic polypeptides comprises the amino acid sequence SEQ ID NO:5
Thr-Pro-Glu-Gly-Ala-Glu of SEQ ID NO:1.

3. The method according to claim 1, wherein the cyclic polypeptide comprises at least seven contiguous amino acids from the amino acid sequence SEQ ID NO: 1.

4. The method according to claim 1, wherein the cyclic polypeptide comprises an entirety of the amino acid sequence SEQ ID NO:1
Gly-Gln-Arg-Glu-Thr-Pro-Glu-Gly-Ala-Glu-Ala-Lys-Pro-Trp-Tyr.

5. The method according to claim 1, wherein the cyclic polypeptide includes a disulfide bond between two cysteine amino acids.

6. The method according to claim 1, wherein the cyclic polypeptide comprises SEQ ID NO:2
Cys-Gly-Gln-Arg-Glu-Thr-Pro-Glu-Gly-Ala-Glu-Ala-Lys-Pro-Trp-Tyr-Cys.

7. The method according to claim 1, wherein the cyclic polypeptide includes a non-natural amino acid.

8. The method according to claim 7, wherein the non-natural amino acid is -aminobutyric acid (GABA).

9. The method according to claim 1, wherein the cyclic polypeptide comprises at least 10 amino acids of SEQ ID NO:1.

10. The method according to claim 1, characterized in that it wherein the cyclic polypeptide is SEQ ID NO:3 ##STR00004## or SEQ ID NO:4 ##STR00005##

11. A pharmaceutical composition for treating autosomal recessive pseudohypoaldosteronism type 1 (PHA type1B) or for restoring the Na.sup.+ ion transport capacity of mutated loss-of-function ENaC, the pharmaceutical composition comprising an effective amount of at least one cyclic polypeptide comprising at least six contiguous amino acids from the amino acid sequence SEQ ID NO:1
Gly-Gln-Arg-Glu-Thr-Pro-Glu-Gly-Ala-Glu-Ala-Lys-Pro-Trp-Tyr.

12. A pharmaceutical composition according to claim 11, further comprising at least one pharmaceutical carrier molecule.

13. A method of treating a patient suffering from autosomal recessive pseudohypoaldosteronism type 1 (PHA type1B) comprising: administering to a patient in need thereof an effective amount of a cyclic polypeptide comprising at least six contiguous amino acids from the amino acid sequence SEQ ID NO:1
Gly-Gln-Arg-Glu-Thr-Pro-Glu-Gly-Ala-Glu-Ala-Lys-Pro-Trp-Tyr.

14. A method for the restoration of the Na.sup.+ transport capacity of a patient with mutant loss-of-function ENaC to physiological level comprising: administering to a patient in need thereof an effective amount of a cyclic polypeptide comprising at least six contiguous amino acids from the amino acid sequence SEQ ID NO:1 Gly-Gln-Arg-Glu-Thr-Pro-Glu-Gly-Ala-Glu-Ala-Lys-Pro-Trp-Tyr.

15. The method according to claim 9, wherein the cyclic polypeptide comprises at least 14 amino acids of SEQ ID NO:1.

Description

DETAILED DESCRIPTION

[0036] Further details of the invention are depicted in the figures and their description.

[0037] FIG. 1 shows a comparison of peptides AP301 and AP318.

[0038] FIG. 2 shows the effect of AP301 on G70S-hENaC. Whole cell I/V relationship of HEK-293 cells transiently expressing G70S-hENaC plotted for control, in presence of 240 nM AP301 and after addition of 10 M amiloride.

[0039] FIG. 3 shows the effect of AP301 on G40S-hENaC. Whole cell I/V relationship of HEK-293 cells transiently expressing G40S-hENaC plotted for control, in presence of 240 nM AP301 and after addition of 10 M amiloride.

[0040] FIG. 4 shows the effect of AP301 on G71S-hENaC. Whole cell I/V relationship of HEK-293 cells transiently expressing G71S-hENaC plotted for control, in presence of 240 nM AP301 and after addition of 10 M amiloride.

[0041] FIG. 5 shows the effect of AP301 on PHA type Ib mutant G37S-hENaC. Whole cell I/V relationship of HEK-293 cells transiently expressing G37S-hENaC plotted for control, in presence of 240 nM AP301 and after addition of 10 M amiloride.

[0042] FIG. 6 shows bar graphs of amiloride-sensitive inward sodium currents. HEK-293 cells transiently transfected with indicated mutant subunits were patched in the whole cell mode; inward current was elicited at 100 mV. 200 nM of AP318 or AP301 were applied.

Synthesis of Cyclic Peptides

[0043] All peptides are synthesised by solid-phase methods; they have been designed to retain the native conformation of the lectin-like domain as much as possible whilst at the same time exploring alternative linking solutions to bring about cyclisation of the linear sequence.

[0044] Peptides are synthesised by solid-phase peptide synthesis according to the fluorenylmethyloxycarbonyl/t-butyl protection strategy on 2-chlorotritylchloride resin. Diisopropyl carbodiimide and N-hydroxybenzotriazole are used as coupling reagents. All coupling steps are carried out in NN-dimethyl formamide. Protected amino acids are coupled in succession to the peptide chain, starting with the C-terminal amino acid. Deprotection of fluorenylmethoxycarbonyl is carried out in 20% piperidine in NN-dimethyl formamide. Cleavage of the completed, partially-protected peptide from the resin is carried out in a 1:1 mixture of acetic acid and dichloromethane. In the case of solnatide and mutant TIP peptide, after cleavage from the resin, side-chain deprotection in 95% trifluoroacetic acid, 5% water, is carried out followed by cyclisation by oxidation of terminal cysteine residues, achieved by aeration of the crude linear peptide at pH 8.5 for 90 hours. Crude peptide product is purified by reverse phase medium pressure liquid chromatography (RP-MPLC) on an RP-C18-silica gel column with a gradient of 5%-40% acetonitrile. Finally, the trifluoracetate counter-ion is replaced by acetate on a Lewatit MP64 column (acetate form). Following a final wash in water, the purified peptide as acetate salt is lyophilised and obtained as a white to off-white powder. In the case of cysteine-free peptides, the cyclisation step is carried out on the partially-protected linear peptide following cleavage from the 2-chlorotritylchloride resin. After selective cyclisation of the cysteine-free peptides, side-chain deprotection in trifluoroacetic acid followed by preparative RP-MPLC, replacement of the trifluoroacetate ion by acetate and lyophilisation of the acetate form of the peptide was carried out as for cysteine-containing peptides. In the case of AP318, in which cyclisation involves amide bond formation through the side chain carboxyl group of aspartic acid, selective cyclisation is achieved by starting the synthesis using the C-terminal aspartic acid N-protected with the fluorenylmethyloxycarbonyl group and with the C-alpha carboxyl protected with a tertiary butyl (OtBu) group. Synthesis proceeds by linkage of the C-terminal aspartic acid residue to the trityl resin through the side chain carboxyl group, followed by stepwise addition of the protected amino acid residues to the peptide chain. After deprotection of the amino group of N-terminal 4-aminobutanoic acid and cleavage of the side-chain protected peptide from the resin, cyclisation is carried out through the free side-chain carboxyl group and the amino group of N-terminal 4-aminobutanoic acid. Finally, side chain protecting groups are removed with trifluoracetic acid and the peptide purified by RP-MPLC as for the other peptides.

[0045] Molecular masses of the peptides are confirmed by electrospray ionisation mass spectrometry or MALDI-TOF-MS and their purity determined by analytical high performance liquid chromatography.

Electrophysiological Assay of AP301 and AP318 Activation of Endogenously and Heterologously-Expressed ENaC

[0046] The ENaC-activating properties of cyclic peptides are tested electrophysiologically in vitro using whole cell and single cell patch clamp techniques Hazemi et al, 2010; Tzotzos et al, 2013; Shabbir et al, 2013). A whole call patch clamp assay is used to measure the induced amiloride-sensitive Na.sup.+ current, calculate concentration-response curves and thus estimate the potency measured as effective concentration at half maximum response (EC50), for cyclic peptides.

[0047] Patch clamp experiments are performed to estimate the potency of AP301/AP318 on transiently expressed -hENaC in HEK-293, CHO cells and A549 cells

Summary of the Rationale for Investigating the Potential Application of AP301 and AP318 for Treatment of Pulmonary Symptoms of Patients Suffering from PHA Type 1b

[0048] Pseudohypoaldosteronism type 1B (PHA type 1B) is caused by loss-of-function mutations in the genes encoding the amiloride-sensitive epithelial sodium channel (ENaC). The condition presents in newborns as life-threatening severe dehydration, hyponatremia and hyperkalaemia due to sodium loss involving kidneys, colon, lungs and sweat and salivary glands; children suffer from pulmonary ailments because reduced sodium-dependent liquid absorption results in elevated lung liquid levels. The disease shows no improvement with age and patients require life-long salt supplements and dietary manipulation to reduce potassium levels.

[0049] AP301 and AP318 can be applied in a patch clamp assay to test the response of cells heterologously-expressing human ENaC subunits into which loss-of-function mutations known to cause PHA type 1B have been introduced by site-directed mutagenesis. In this way the ability of AP301 and AP318 to restore the amiloride-sensitive sodium current in cells expressing these mutated loss-of-function ENaC subunits, can be measured. Increase in the sodium current in the presence of cyclic peptides indicates their ability to restore Na.sup.+ ion movement by loss-of-function ENaC carrying PHA type 1B mutations, to restore ENaC function therefore and their potential as therapies for PHA type 1B patients.

[0050] Surprisingly it has been detected that cyclic peptides such as AP301 and AP318 can restore Na.sup.+ ion transport activity to loss-of-function mutant ENaC. Thus AP301 and AP318 are potential therapies for pulmonary symptoms of PHA type IB.

Experimental Protocol

In Vitro Study of Effect of AP301 and AP318 on Heterologously Expressed ENaC Carrying PHA Type IB Mutations

[0051] The effect of cyclic peptides on the amiloride-sensitive Na.sup.+ current was observed in HEK cells heterologously expressing human ENaC subunits (HEK cells show no endogenous expression of ENaC [Ruffieux-Daidie et al, 2008]) into which single point mutations of alpha, beta and gamma ENaC, the same as those found responsible for the pathological phenotype of patients suffering from PHA type IB, had been introduced by site-directed mutagenesis. In addition, mutant delta ENaC subunits were also constructed by site-directed mutagenesis, containing homologous mutations to those observed in conserved positions in the other three ENaC subunits.

[0052] Construction of ENaC PHA Type I Mutants and Expression in HEK 293

[0053] Various types of mutations of ENaC can be reproduced by site directed mutagenesis of wild type ENaC subunit DNA cloned into plasmid vectors.

[0054] Site-Directed Mutagenesis

[0055] Point mutations were introduced into cDNA encoding alpha, beta, gamma and delta ENaC using a commercially available site-directed mutagenesis kit (QuikChange Lightning Site-Directed Mutagenesis Kit; Agilent Technologies). The cDNAs encoding alpha, beta, and gamma-hENaC had been donated by Dr. Peter Snyder (University of Iowa, Carver College of Medicine, Iowa City, Iowa); cDNA encoding delta-hENaC had been donated by Dr. Mike Althaus (Justus-Liebig University, Giessen, Germany).

[0056] Mutagenic primers were designed individually based on descriptions of the individual mutations in the original scientific reports. The primer design program provided on the manufacturer's website was used as a guide and primers themselves were ordered from Sigma-Aldrich. Mutant strands were synthesised by PCR with a Pfu-based DNA polymerase using 100 ng wild-type (WT) cDNA encoding alpha, beta, gamma or delta-hENaC. Parental (WT) strands were removed and the resulting plasmid DNA containing mutated ENaC was transformed into E. coli competent cells. Following growth in culture, plasmid DNA was extracted from the E. coli cells using a commercially available plasmid isolation kit (GeneJET Plasmid Miniprep Kit; Thermoscientific) and isolated by column chromatography. All the mutant constructs were checked by restriction site mapping and sequencing.

[0057] Transfection of HEK-293 Cells for Heterologous Expression of hENaC

[0058] HEK-293 cells were transfected with the mutant alpha-, beta-, gamma- and delta-hENaC and WT alpha-, beta-, gamma- and delta-hENaC plasmid DNA using a commercially available kit (X-treme Gene HP transfection reagent (Roche Diagnostics, Mannheim, Germany) following the protocol recommended by the manufacturer. One mutant subunit together with the remaining two WT subunits were transfected simultaneously to give expression of trimeric mutant ENaC. Expression of WT ENaC was achieved by simultaneous transfection with WT alpha-, beta- and gamma-hENaC plasmid DNA or with WT delta-, beta- and gamma-hENaC plasmid DNA.

[0059] Patch Clamp Testing of ENaC-Activating Ability of AP301 and AP318 in HEK Cells Transiently Expressing Mutant ENaC

[0060] Each HEK-293 cell line transiently expressing WT -hENaC, WT -hENaC or a mutant hENaC subunit co-expressed with WT subunits, was tested in a whole cell patch clamp assay with AP301 and selected mutants at conserved positions with AP318. Whole cell currents were recorded as previously described (Shabbir et al, 2013).

[0061] Concentration Response Measurements

[0062] Concentration-response curves were plotted, and EC50 values and Hill coefficients were determined using Microcal Origin 7.0. The whole-cell sodium current of HEK-293 cells transiently transfected with WT -hENaC, WT -hENaC or a mutant hENaC, was recorded at a holding potential (Eh) of 80 mV following cumulative addition of AP301 stock solution to the bath solution, resulting in final concentrations ranging from 3.5 to 240 nM solnatide. Finally, amiloride was added to enable estimation of the peptide-induced increase in amiloride-sensitive Na+ current. The activity of AP301 was expressed as a percentage of the paired amiloride response, owing to variability in hENaC expression between different batches of cultured cells. Amiloride was used at 10 M for WT -hENaC, WT -hENaC or mutant hENaC; these concentrations yielded greater than 95% hENaC inhibition. Only cells with clear amiloride response were included in data analysis.

[0063] Current-Voltage Relationships

[0064] Whole-cell current-voltage (I/V) relationships of HEK-293 cells transiently infected with -hENaC, WT -hENaC or a mutant hENaC were determined for control (before addition of AP301), treatment with 240 nM AP301 and following addition of 10 M amiloride, respectively. After GOhm-seal (G-seal) formation, and an equilibration period of 5 min, sodium current was recorded at Eh from 80 to +80 mV in 20 mV increments held for 1 min at each Eh.

[0065] Statistical Analysis

[0066] Data represent the meanS.E. unless otherwise stated; experiments were performed on three to seven batches of independently transfected cells in the HEK-293 heterologous expression system. Statistical significance between different groups was determined using an unpaired, two-tailed Student's t test using GraphPad Prism version 3.02 (GraphPad Software, San Diego).

Results

[0067]

TABLE-US-00001 TABLE 1 Results of AP301 in a whole cell patch clamp assay with HEK- 293 cells expressing PHA type IB mutant hENaC and homologues Amiloride-sensitive AP301 restored current Na+ current EC.sub.50 hENaC (pA) (pA) (nM) WT 81.2 5.5 54.7 2.2.sup.c WT 93.5 9.5 46.2 1.5 G70S loss-of-function 392.1 14.2 61.9 2.1 G37S.sup.a loss-of-function 192.8 12.3 65.8 3.2 G40S loss-of-function 90.8 8.9 69.1 2.7 G71S loss-of-function 185.1 21.2 42.9 0.5 Q101K.sup.a loss-of-function 79.1 11.2 Q66K loss-of-function 448.5 57.5 56.9 16.7 Q70K loss-of-function 305.2 10.1 61.5 4.0 Q102K loss-of-function 121.9 14.6 C133Y.sup.a loss-of-function 111.8 10.6 C98Y loss-of-function 296.3 4.2 79.4 1.7 C100Y loss-of-function 307.9 15.7 88.2 11.9 C134Y loss-of-function 219.1 19.8 G327C.sup.a loss-of-function 196.8 16.7 G294C loss-of-function 76.9 13.5 G305C loss-of-function 125.5 15.7 G303C loss-of-function 48.5 8.2 (.sup.amutants observed in patients)

[0068] Effect of AP301 on PHA Type IB G37S-hENaC and Homologues

[0069] The effect of AP301 on HEK-293 cells expressing the PHA type 1b mutant G37S-hENaC or one of its lab-constructed homologues is shown in Figures below. Whole-cell current-voltage (I/V) relationships are shown for each mutant loss-of-function hENaC transiently expressed in HEK-293 cells as well as absolute mean values of inward current density at a holding potential of 80 mV during control phase, following addition of 240 nM AP301 and after final addition of amiloride (10 M) to the bath solution (Figures below).

[0070] Effect of AP318 on PHA Type IB Mutant ENaC

[0071] In order to test whether the activity restoring effect on PHA type 1B mutant ENaC is exclusive to AP301 or whether it is a property of cyclic peptides in general, three PHA type IB mutant ENaCs containing mutations in the -, - and -hENaC subunits, respectively, were tested in a whole cell patch clamp assay in the presence of AP318 as well AP301. All three mutants have been observed in patients suffering from PHA type 1B and occur at conserved positions in the ENaC subunits; two of these mutants, Q101K-hENaC and G37S-hENaC had been previously tested with AP301. The third mutant occurs in the subunit: V543fs-hENaC and in contrast to all mutants so far tested is a frameshift mutant resulting in a truncated subunit.

[0072] The results of testing HEK-293 cells expressing these mutant ENaCs in a whole cell patch clamp assay in the presence of AP318 and AP301 are shown in Table 2 and FIG. 6.

TABLE-US-00002 TABLE 2 Amiloride-sensitive current in a whole cell patch clamp assay of HEK-293 cells transiently expressing WT ENaC and ENaC containing PHA type 1b mutations in the absence (Control) and presence of AP301 and AP318 (mean SE values for inward current measured in pA, n = 5; peptide concentration 220 nM). No peptide AP301 AP318 Inward current Inward current measured (pA) ENaC (pA) Peptide concentration 220 nM WT -hENaC 67.2 5.2 Q101K-hENaC loss-of-function 174.7 8.7 176.7 6.9 G37S-hENaC loss-of-function 189.8 7.1 191.8 5.8 V543fs-hENaC loss-of-function 143.3 8.2 177.7 6.5

[0073] The Following Conclusions can be Drawn from the Results Obtained so Far:

[0074] 1) PHA type IB mutations resulted in a loss-of-function in the amiloride-sensitive sodium current through ENaC compared to WT hENaC.

[0075] 2) AP301 restored Na.sup.+ ion transport capacity and compensated for amino acid mutations of all the mutant loss-of-function hENaCs observed in PHA type IB patients: G37S-, Q101K- and G327C-hENaC.

[0076] 3) Compared to the physiological level of amiloride-sensitive sodium ion current observed with WT - and -hENaC, AP301 restored amiloride-sensitive sodium ion current of mutant loss-of-function ENaC to levels comparable to non-mutant active ENaC.

[0077] 4) Concentration-response curves and EC50 values for PHA type IB mutant G37S-hENaC and corresponding homologues indicate that AP301 has the potential to restore activity and to compensate amino acid mutations in all of these mutant ENaC channels, which have loss-of-function compared to WT - and -hENaC

[0078] 5) Current-voltage (IN) relationships for PHA type IB mutant G37S-hENaC and corresponding homologues characterise the restoring effect of AP301 on these mutant channels in greater detail. The same mutation occurring at a conserved position in the different subunits of hENaC, results in an sodium ion channel with different functional properties and phenotypic effect, as clearly reflected by the restored activity in the presence of AP301 (Table 3).

[0079] 6) Results of the whole cell electrophysiological assay in the absence of AP301 and AP318 show that, compared to wild type, PHA type 1B mutations in all -hENaC subunits result in loss-of-function. In the presence of AP301 and AP318, significantly increased amiloride-sensitive sodium ion current through PHA-1B mutants demonstrating restoration of normal sodium ion channel function was observed.

OVERALL CONCLUSION

[0080] It can be concluded from these results that AP301 and AP318 can restore Na.sup.+ ion transport and compensate for amino acid mutations in loss-of-function PHA type IB mutant hENaCs, indicating the potential of cyclic peptides to restore impaired ENaC function and to compensate for amino acid mutations in these mutants and thereby act as a therapy to treat patients suffering from systemic PHA type I.

REFERENCES

[0081] 1. Aberer E, Gebhart W, Mainitz M, Pollak A, Reichel G, Scheibenreiter S. [Sweat glands in pseudohypoaldosteronism]. Hautarzt. 1987 August; 38(8):484-7. German. PubMed PMID: 3654220. [0082] 2. Adachi M, Tachibana K, Asakura Y, Abe S, Nakae J, Tajima T, Fujieda K. Compound heterozygous mutations in the gamma subunit gene of ENaC (1627delG and 1570-1G-->A) in one sporadic Japanese patient with a systemic form of pseudohypoaldosteronism type 1. J Clin Endocrinol Metab. 2001 January; 86(1):9-12. PubMed PMID: 11231969. [0083] 3. Akkurt I, Kuhnle U, Ringenberg C. Pseudohypo-aldosteronism and cholelithiasis: coincidence or pathogenetic correlation? Eur J Pediatr. 1997 May; 156(5):363-6. PubMed PMID: 9177977. [0084] 4. Althaus M, Clauss W G, Fronius M. Amiloride-sensitive sodium channels and pulmonary edema. Pulm Med. 2011; 2011:830320. Epub 2010 Dec. 29. PubMed PMID: 21637371. [0085] 5. Amin N, Alvi N S, Barth J H, Field H P, Finlay E, Tyerman K, Frazer S, Savill G, Wright N P, Makaya T, Mushtaq T. Pseudohypoaldosteronism type 1: clinical features and management in infancy. Endocrinol Diabetes Metab Case Rep. 2013; 2013:130010. doi: 10.1530/EDM-13-0010. Epub 2013 Aug. 30. PubMed PMID: 24616761; PubMed Central PMCID: PMC3922296. [0086] 6. Baconguis I, Bohlen C J, Goehring A, Julius D, Gouaux E. X-ray structure of acid-sensing ion channel 1 snake toxin complex reveals open state of a Na(+)-selective channel. Cell. 2014 Feb. 13; 156(4):717-29. doi: 10.1016/j.cell.2014.01.011. Epub 2014 Feb. 6. PubMed PMID: 24507937; PubMed Central PMCID: PMC4190031. [0087] 7. Belot A, Ranchin B, Fichtner C, Pujo L, Rossier B C, Liutkus A, Morlat C, Nicolino M, Zennaro M C, Cochat P. Pseudohypoaldosteronisms, report on a 10-patient series. Nephrol Dial Transplant. 2008 May; 23(5):1636-41. doi: 10.1093/ndt/gfm862. PubMed PMID: 18424465. [0088] 8. Bonny O, Chraibi A, Loffing J, Jaeger N F, Grunder S, Horisberger J D, Rossier B C. Functional expression of a pseudohypoaldosteronism type I mutated epithelial Na.sup.+ channel lacking the pore-forming region of its alpha subunit. J Clin Invest. 1999 October; 104(7):967-74. PubMed PMID: 10510337; PubMed Central PMCID: PMC408554. [0089] 9. Bonny O, Knoers N, Monnens L, Rossier B C. A novel mutation of the epithelial Na.sup.+ channel causes type 1 pseudohypoaldosteronism. Pediatr Nephrol. 2002 October; 17(10):804-8. Epub 2002 Aug. 21. PubMed PMID: 12376807. [0090] 10. Bonny O, Rossier B C. Disturbances of Na/K balance: pseudohypoaldosteronism revisited. J Am Soc Nephrol. 2002 September; 13(9):2399-414. Review. PubMed PMID: 12191985. [0091] 11. Braun C, Hamacher J, Morel D R, Wendel A, Lucas R. Dichotomal role of TNF in experimental pulmonary edema reabsorption. J Immunol. 2005 Sep. 1; 175(5):3402-8. PubMed PMID: 16116234. [0092] 12. Canessa C M, Schild L, Buell G, Thorens B, Gautschi I, Horisberger J D, Rossier B C. (B) Amiloride-sensitive epithelial Na.sup.+ channel is made of three homologous subunits. Nature. 1994 Feb. 3; 367(6462):463-7. PubMed PMID: 8107805. [0093] 13. Chang S S, Grunder S, Hanukoglu A, Rosier A, Mathew P M, Hanukoglu I, Schild L, Lu Y, Shimkets R A, Nelson-Williams C, Rossier B C, Lifton R P. Mutations in subunits of the epithelial sodium channel cause salt wasting with hyperkalaemic acidosis, pseudohypoaldosteronism type 1. Nat Genet. 1996 March; 12(3):248-53. PubMed PMID: 8589714. [0094] 14. Cheek D B, Perry J W. A salt wasting syndrome in infancy. Arch Dis Child. 1958 June; 33(169):252-6. PubMed PMID: 13545877; PubMed Central PMCID: PMC2012226. [0095] 15. Czikora I, Alli A, Bao H F, Kaftan D, Sridhar S, Apell H J, Gorshkov B, White R, A, Wendel A, Pauly-Evers M, Hamacher J, Garcia-Gabay I, Fischer B, Verin A, Bagi Z, Pittet J F, Shabbir W, Lemmens-Gruber R, Chakraborty T, Lazrak A, Matthay M A, Eaton D C, Lucas R. A Novel TNF-mediated Mechanism of Direct Epithelial Sodium Channel Activation. Am J Respir Crit Care Med. 2014 Jul. 16. PubMed PMID: 25029038. [0096] 16. Dirlewanger M, Huser D, Zennaro M C, Girardin E, Schild L, Schwitzgebel V M. A homozygous missense mutation in SCNN1A is responsible for a transient neonatal form of pseudohypoaldosteronism type 1. Am J Physiol Endocrinol Metab. 2011 September; 301(3):E467-73. doi: 10.1152/ajpendo.00066.2011. Epub 2011 Jun. 7. PubMed PMID: 21653223. [0097] 17. Dogan C S, Erdem D, Mesut P, Merve A, Sema A, Iffet B, Afig B. A novel splice site mutation of the beta subunit gene of epithelial sodium channel (ENaC) in one Turkish patient with a systemic form of pseudohypoaldosteronism Type 1. J Pediatr Endocrinol Metab. 2012; 25(9-10):1035-9. doi: 10.1515/jpem-2012-0083. PubMed PMID: 23426840. [0098] 18. Dulebo A, Ettrich R, Lucas R, Kaftan D. A computational study of the oligosaccharide binding sites in the lectin-like domain of Tumor Necrosis Factor and the TNF-derived TIP peptide. Curr Pharm Des. 2012; 18(27):4236-43. PubMed PMID: 22697478; PubMed Central PMCID: PMC3495565. [0099] 19. Eaton D C, Helms M N, Koval M, Bao H F, Jain L. The contribution of epithelial sodium channels to alveolar function in health and disease. Annu Rev Physiol. 2009; 71:403-23. doi: 10.1146/annurev.physiol.010908.163250. Review. PubMed PMID: 18831683. [0100] 20. Edelheit O, Hanukoglu I, Gizewska M, Kandemir N, Tenenbaum-Rakover Y, Yurdakok M, Zajaczek S, Hanukoglu A. Novel mutations in epithelial sodium channel (ENaC) subunit genes and phenotypic expression of multisystem pseudohypoaldosteronism. Clin Endocrinol (Oxf). 2005 May; 62(5):547-53. Review. PubMed PMID: 15853823. [0101] 21. Edelheit O, Hanukoglu I, Shriki Y, Tfilin M, Dascal N, Gillis D, Hanukoglu A. Truncated beta epithelial sodium channel (ENaC) subunits responsible for multi-system pseudohypoaldosteronism support partial activity of ENaC. J Steroid Biochem Mol Biol. 2010 March; 119(1-2):84-8. doi: 10.1016/j.jsbmb.2010.01.002. Epub 2010 Jan. 12. PubMed PMID: 20064610. [0102] 22. Ekinci Z, Aytac M B, Cheong H I. A case of SCNN1A splicing mutation presenting as mild systemic pseudohypoaldosteronism type 1. J Pediatr Endocrinol Metab. 2013; 26(11-12):1197-200. doi: 10.1515/jpem-2013-0053. PubMed PMID: 23813355. [0103] 23. Elia N, Tapponnier M, Matthay M A, Hamacher J, Pache J C, Brundler M A, Totsch M, De Baetselier P, Fransen L, Fukuda N, Morel D R, Lucas R. Functional identification of the alveolar edema reabsorption activity of murine tumor necrosis factor-alpha. Am J Respir Crit Care Med. 2003 Nov. 1; 168(9):1043-50. Epub 2003 Jul. 3. PubMed PMID: 12842853. [0104] 24. Fukuda N, Jayr C, Lazrak A, Wang Y, Lucas R, Matalon S, Matthay M A. Mechanisms of TNF-alpha stimulation of amiloride-sensitive sodium transport across alveolar epithelium. Am J Physiol Lung Cell Mol Physiol. 2001 June; 280(6):L1258-65. PubMed PMID: 11350806. [0105] 25. Garry H, Palmer L G. Epithelial sodium channels: function, structure, and regulation. Physiol Rev. 1997 April; 77(2):359-96. Review. PubMed PMID: 9114818. [0106] 26. Geller D S, Rodriguez-Soriano J, Vallo Boado A, Schifter S, Bayer M, Chang S S, Lifton R P. Mutations in the mineralocorticoid receptor gene cause autosomal dominant pseudohypoaldosteronism type I. Nat Genet. 1998 July; 19(3):279-81. PubMed PMID: 9662404. [0107] 27. Gonzales E B, Kawate T, Gouaux E. Pore architecture and ion sites in acid-sensing ion channels and P2X receptors. Nature. 2009 Jul. 30; 460(7255):599-604. doi: 10.1038/nature08218. PubMed PMID: 19641589; PubMed Central PMCID: PMC2845979. [0108] 28. Grnder S, Firsov D, Chang S S, Jaeger N F, Gautschi I, Schild L, Lifton R P, Rossier B C. A mutation causing pseudohypoaldosteronism type 1 identifies a conserved glycine that is involved in the gating of the epithelial sodium channel. EMBO J. 1997 Mar. 3; 16(5):899-907. PubMed PMID: 9118951; PubMed Central PMCID: PMC1169690. [0109] 29. Gran T, Degirmenci S, Bulut K, Say A, Riepe F G, Gran . Critical points in the management of pseudohypoaldosteronism type 1. J Clin Res Pediatr Endocrinol. 2011; 3(2):98-100. doi: 10.4274/jcrpe.v3i2.20. Epub 2011 Jun. 8. PubMed PMID: 21750640; PubMed Central PMCID: PMC3119449. [0110] 30. Hamacher J, Stammberger U, Roux J, Kumar S, Yang G, Xiong C, Schmid R A, Fakin R M, Chakraborty T, Hossain H M, Pittet J F, Wendel A, Black S M, Lucas R. The lectin-like domain of tumor necrosis factor improves lung function after rat lung transplantationpotential role for a reduction in reactive oxygen species generation. Crit Care Med. 2010 March; 38(3):871-8. PubMed PMID: 20081530. [0111] 31. Hanukoglu A, Bistritzer T, Rakover Y, Mandelberg A. Pseudohypoaldosteronism with increased sweat and saliva electrolyte values and frequent lower respiratory tract infections mimicking cystic fibrosis. J Pediatr. 1994 November; 125(5 Pt 1):752-5. PubMed PMID: 7965429. [0112] 32. Hanukoglu A, Edelheit O, Shriki Y, Gizewska M, Dascal N, Hanukoglu I. Renin-aldosterone response, urinary Na/K ratio and growth in pseudohypoaldosteronism patients with mutations in epithelial sodium channel (ENaC) subunit genes. J Steroid Biochem Mol Biol. 2008 September; 111(3-5):268-74. doi: 10.1016/j.jsbmb.2008.06.013. Epub 2008 Jun. 26. PubMed PMID: 18634878. [0113] 33. Hartmann E K, Boehme S, Duenges B, Bentley A, Klein K U, Kwiecien R, Shi C, Szczyrba M, David M, Markstaller K. An inhaled tumor necrosis factor-alpha-derived TIP peptide improves the pulmonary function in experimental lung injury. Acta Anaesthesiol Scand. 2013 March; 57(3):334-41. doi: 10.1111/aas.12034. Epub 2012 Dec. 6. PubMed PMID: 23216436. [0114] 34. Hartmann E K, Thomas R, Liu T, Stefaniak J, Ziebart A, Duenges B, Eckle D, Markstaller K, David M. TIP peptide inhalation in experimental acute lung injury: effect of repetitive dosage and different synthetic variants. BMC Anesthesiol. 2014 May 26; 14:42. doi: 10.1186/1471-2253-14-42. eCollection 2014. PubMed PMID: 24904234; PubMed Central PMCID: PMC4046002. [0115] 35. Hazemi P, Tzotzos S J, Fischer B, Andavan G S, Fischer H, Pietschmann H, Lucas R, Lemmens-Gruber R. Essential structural features of TNF- lectin-like domain derived peptides for activation of amiloride-sensitive sodium current in A549 cells. J Med Chem. 2010 Nov. 25; 53(22):8021-9. doi: 10.1021/jm100767p. Epub 2010 Oct. 27. PubMed PMID: 20979368; PubMed Central PMCID: PMC2996585. [0116] 36. Hribar M, Bloc A, van der Goot F G, Fransen L, De Baetselier P, Grau G E, Bluethmann H, Matthay M A, Dunant Y, Pugin J, Lucas R. The lectin-like domain of tumor necrosis factor-alpha increases membrane conductance in microvascular endothelial cells and peritoneal macrophages. Eur J Immunol. 1999 October; 29(10):3105-11. PubMed PMID: 10540321. [0117] 37. Huber R, Krueger B, Diakov A, Korbmacher J, Haerteis S, Einsiedel J, Gmeiner P, Azad A K, Cuppens H, Cassiman J J, Korbmacher C, Rauh R. Functional characterization of a partial loss of-function mutation of the epithelial sodium channel (ENaC) associated with atypical cystic fibrosis. Cell Physiol Biochem. 2010; 25(1):145-58. doi: 10.1159/000272059. Epub 2009 Dec. 22. PubMed PMID: 20054153. [0118] 38. Hummler E, Barker P, Gatzy J, Beermann F, Verdumo C, Schmidt A, Boucher R, Rossier B C. Early death due to defective neonatal lung liquid clearance in alpha-ENaC-deficient mice. Nat Genet. 1996 March; 12(3):325-8. PubMed PMID: 8589728. [0119] 39. Huppmann S, Lankes E, Schnabel D, Bhrer C. Unimpaired postnatal respiratory adaptation in a preterm human infant with a homozygous ENaC- unit loss-of-function mutation. J Perinatol. 2011 December; 31(12):802-3. doi: 10.1038/jp.2011.46. PubMed PMID: 22124517. [0120] 40. Jain L, Chen X J, Ramosevac S, Brown L A, Eaton D C. (2001) Expression of highly selective sodium channels in alveolar type II cells is determined by culture conditions. Am J Physiol Lung Cell Mol Physiol. 280:L646-58. [0121] 41. Jasti J, Furukawa H, Gonzales E B, Gouaux E. Structure of acid-sensing ion channel 1 at 1.9 A resolution and low pH. Nature. 2007 Sep. 20; 449(7160):316-23. PubMed PMID: 17882215. [0122] 42. Kala Ahluwalia G, Dasouki M, Lennon A. Phenotypic variation of autosomal recessive pseudohypoaldosteronism type I: a case in point. Clin Case Rep. 2014 December; 2(6):326-30. doi: 10.1002/ccr3.129. Epub 2014 Sep. 15. PubMed PMID: 25548639; PubMed Central PMCID: PMC4270719. [0123] 43. Kashlan O B, Adelman J L, Okumura S, Blobner B M, Zuzek Z, Hughey R P, Kleyman T R, Grabe M. Constraint-based, homology model of the extracellular domain of the epithelial Na+ channel subunit reveals a mechanism of channel activation by proteases. J Biol Chem. 2011 Jan. 7; 286(1):649-60. doi: 10.1074/jbc.M110.167098. Epub 2010 Oct. 25. PubMed PMID: 20974852; PubMed Central PMCID: PMC3013024. [0124] 44. Kellenberger S, Schild L. Epithelial sodium channel/degenerin family of ion channels: a variety of functions for a shared structure. Physiol Rev. 2002 July; 82(3):735-67. Review. PubMed PMID: 12087134. [0125] 45. Kerem E, Bistritzer T, Hanukoglu A, Hofmann T, Zhou Z, Bennett W, MacLaughlin E, Barker P, Nash M, Quittell L, Boucher R, Knowles M R. Pulmonary epithelial sodium-channel dysfunction and excess airway liquid in pseudohypoaldosteronism. N Engl J Med. 1999 Jul. 15; 341(3):156-62. PubMed PMID: 10403853. [0126] 46. Kosari F, Sheng S, Li J, Mak D O, Foskett J K, Kleyman T R. Subunit stoichiometry of the epithelial sodium channel. J Biol Chem. 1998 May 29; 273(22):13469-74. PubMed PMID: 9593680. [0127] 47. Krueger B, Schlotzer-Schrehardt U, Haerteis S, Zenkel M, Chankiewitz V E, Amann K U, Kruse F E, Korbmacher C. Four subunits () of the epithelial sodium channel (ENaC) are expressed in the human eye in various locations. Invest Ophthalmol Vis Sci. 2012 Feb. 2; 53(2):596-604. doi: 10.1167/iovs.11-8581. PubMed PMID: 22167092. [0128] 48. Lazrak A, Samanta A, Venetsanou K, Barbry P, Matalon S. Modification of biophysical properties of lung epithelial Na(+) channels by dexamethasone. Am J Physiol Cell Physiol. 2000 September; 279(3):C762-70. PubMed PMID: 10942727. [0129] 49. Lifton R P, Gharavi A G, Geller D S. Molecular mechanisms of human hypertension. Cell. 2001 Feb. 23; 104(4):545-56. Review. PubMed PMID: 11239411. [0130] 50. Lingueglia E, Voilley N, Waldmann R, Lazdunski M, Barbry P. Expression cloning of an epithelial amiloride-sensitive Na+ channel. A new channel type with homologies to Caenorhabditis elegans degenerins. FEBS Lett. 1993 Feb. 22; 318(1):95-9. PubMed PMID: 8382172. [0131] 51. Lucas R, Magez S, De Leys R, Fransen L, Scheerlinck J P, Rampelberg M, Sablon E, De Baetselier P. Mapping the lectin-like activity of tumor necrosis factor. Science. 1994 Feb. 11; 263(5148):814-7. PubMed PMID: 8303299. [0132] 52. Lucas R, Yang G, Gorshkov B A, Zemskov E A, Sridhar S, Umapathy N S, Jezierska-Drutel A, Alieva I B, Leustik M, Hossain H, Fischer B, Catravas J D, Verin A D, Pittet J F, Caldwell R B, Mitchell T J, Cederbaum S D, Fulton D J, Matthay M A, Caldwell R W, Romero M J, Chakraborty T. Protein kinase C- and arginase I mediate pneumolysin-induced pulmonary endothelial hyperpermeability. Am J Respir Cell Mol Biol. 2012 October; 47(4):445-53. doi: 10.1165/rcmb.2011-03320C. Epub 2012 May 10. PubMed PMID: 22582175; PubMed Central PMCID: PMC3488628. [0133] 53. Marquardt A, Bernevic B, Przybylski M. Identification, affinity characterisation and biological interactions of lectin-like peptide-carbohydrate complexes derived from human TNF-alpha using high-resolution mass spectrometry. J Pept Sci. 2007 December; 13(12):803-10. PubMed PMID: 17918767. [0134] 54. Marthinsen L, Komflt R, Aili M, Andersson D, Westgren U, Schaedel C. Recurrent Pseudomonas bronchopneumonia and other symptoms as in cystic fibrosis in a child with type I pseudohypoaldosteronism. Acta Paediatr. 1998 April; 87(4):472-4. PubMed PMID: 9628311. [0135] 55. McDonald F J, Yang B, Hrstka R F, Drummond H A, Tarr D E, McCray P B Jr, Stokes J B, Welsh M J, Williamson R A. Disruption of the beta subunit of the epithelial Na+ channel in mice: hyperkalaemia and neonatal death associated with a pseudohypoaldosteronism phenotype. Proc Natl Acad Sci USA. 1999 Feb. 16; 96(4):1727-31. PubMed PMID: 9990092; PubMed Central PMCID: PMC15575. [0136] 56. Mora-Lopez F, Bernal-Quiros M, Lechuga-Sancho A M, Lechuga-Campoy J L, Hernandez-Trujillo N, Nieto A. Novel mutation in the epithelial sodium channel causing type I pseudohypoaldosteronism in a patient misdiagnosed with cystic fibrosis. Eur J Pediatr. 2012 June; 171(6):997-1000. doi: 10.1007/s00431-012-1697-5. Epub 2012 Feb. 28. PubMed PMID: 22371258. [0137] 57. Riepe F G, van Bemmelen M X, Cachat F, Plendl H, Gautschi I, Krone N, Holterhus P M, Theintz G, Schild L. Revealing a subclinical salt-losing phenotype in carriers of the novel S562P mutation in the alpha subunit of the epithelial sodium channel. Clin Endocrinol (Oxf). 2009 February; 70(2):252-8. doi: 10.1111/j.1365-2265.2008.03314.x. PubMed PMID: 18547339. [0138] 58. Riepe F G. Clinical and molecular features of type 1 pseudohypoaldosteronism. Horm Res. 2009; 72(1):1-9. doi: 10.1159/000224334. Epub 2009 Jun. 30. Review. PubMed PMID: 19571553. [0139] 59. Rossier B C, Canessa C M, Schild L, Horisberger J D. Epithelial sodium channels. Curr Opin Nephrol Hypertens. 1994 September; 3(5):487-96. Review. PubMed PMID: 7804746. [0140] 60. Ruffieux-Daidi D, Poirot O, Boulkroun S, Verrey F, Kellenberger S, Staub O. Deubiquitylation regulates activation and proteolytic cleavage of ENaC. J Am Soc Nephrol. 2008 November; 19(11):2170-80. doi: 10.1681/ASN.2007101130. Epub 2008 Aug. 13. PubMed PMID: 18701608; PubMed Central PMCID: PMC2573013. [0141] 61. Saravanapandian N, Paul S, Matthai J. Pseudohypoaldosteronism type 1: a rare cause of severe dyselectrolytemia and cardiovascular collapse in neonates. J Clin Neonatol. 2012 October; 1(4):224-6. doi: 10.4103/2249-4847.106007. PubMed PMID: 24027733; PubMed Central PMCID: PMC3762049. [0142] 62. Saxena A, Hanukoglu I, Saxena D, Thompson R J, Gardiner R M, Hanukoglu A. Novel mutations responsible for autosomal recessive multisystem pseudohypoaldosteronism and sequence variants in epithelial sodium channel alpha-, beta-, and gamma-subunit genes. J Clin Endocrinol Metab. 2002 July; 87(7):3344-50. PubMed PMID: 12107247. [0143] 63. Schaedel C, Marthinsen L, Kristoffersson A C, Komfalt R, Nilsson K O, Orlenius B, Holmberg L. Lung symptoms in pseudohypoaldosteronism type 1 are associated with deficiency of the alpha-subunit of the epithelial sodium channel. J Pediatr. 1999 December; 135(6):739-45. PubMed PMID: 10586178. [0144] 64. Shabbir W, Scherbaum-Hazemi P, Tzotzos S, Fischer B, Fischer H, Pietschmann H, Lucas R, Lemmens Gruber R. Mechanism of action of novel lung edema therapeutic AP301 by activation of the epithelial sodium channel. Mol Pharmacol. 2013 December; 84(6):899-910. doi: 10.1124/mol.113.089409. Epub 2013 Sep. 27. PubMed PMID: 24077967; PubMed Central PMCID: PMC3834145. [0145] 65. Sharma R, Pandey M, Kanwal S K, Zennaro M C. Pseudohypoaldosteronism type 1: management issues. Indian Pediatr. 2013 March; 50(3):331-3. PubMed PMID: 23680607. [0146] 66. Silva N, Costa M, Silva A, S. C, Martins S, Antunes A, Marques O, Castedo S, Pereira A. A case of systemic pseudohypoaldosteronism with a novel mutation in the SCNN1A gene. Endocrinol Nutr. 2013 January; 60(1):33-6. doi: 10.1016/j.endonu.2012.07.002. Epub 2012 Sep. 30. PubMed PMID: 23031435. [0147] 67. Staruschenko A. Regulation of transport in the connecting tubule and cortical collecting duct. Compr Physiol. 2012 April; 2(2):1541-84. Review. PubMed PMID: 23227301; PubMed Central PMCID: PMC3516049. [0148] 68. Stockand J D, Staruschenko A, Pochynyuk O, Booth R E, Silverthorn D U. Insight toward epithelial Na+ channel mechanism revealed by the acid-sensing ion channel 1 structure. IUBMB Life. 2008 September; 60(9):620-8. doi: 10.1002/iub.89. Review. PMID: 18459164. [0149] 69. Strautnieks S S, Thompson R J, Gardiner R M, Chung E. A novel splice-site mutation in the gamma subunit of the epithelial sodium channel gene in three pseudohypoaldosteronism type 1 families. Nat Genet. 1996 June; 13(2):248-50. PubMed PMID: 8640238. [0150] 70. Strautnieks S S, Thompson R J, Hanukoglu A, Dillon M J, Hanukoglu I, Kuhnle U, Seckl J, Gardiner R M, Chung E. Localisation of pseudohypoaldosteronism genes to chromosome 16p12.2-13.11 and 12p13.1-pter by homozygosity mapping. Hum Mol Genet. 1996 February; 5(2):293-9. PubMed PMID: 8824886. [0151] 71. Thomas C P, Zhou J, Liu K Z, Mick V E, MacLaughlin E, Knowles M. Systemic pseudohypoaldosteronism from deletion of the promoter region of the human Beta epithelial na(+) channel subunit. Am J Respir Cell Mol Biol. 2002 September; 27(3):314-9. PubMed PMID: 12204893. [0152] 72. Tzotzos S, Fischer B, Fischer H, Pietschmann H, Lucas R, Dupr G, Lemmens-Gruber R, Hazemi P, Prymaka V, Shabbir W. AP301, a synthetic peptide mimicking the lectin-like domain of TNF, enhances amiloride-sensitive Na(+) current in primary dog, pig and rat alveolar type II cells. Pulm Pharmacol Ther. 2013 June; 26(3):356-63. doi:10.1016/j.pupt.2012.12.011. Epub 2013 Jan. 9. PubMed PMID: 23313096; PubMed Central PMCID: PMC3646188. [0153] 73. Vadsz I, Schermuly R T, Ghofrani H A, Rummel S, Wehner S, Mhldorfer I, Schafer K P, Seeger W, Morty R E, Grimminger F, Weissmann N. The lectin-like domain of tumor necrosis factor alpha improves alveolar fluid balance in injured isolated rabbit lungs. Crit Care Med. 2008 May; 36(5):1543 50. PubMed PMID: 18434905. [0154] 74. Waldmann R, Champigny G, Bassilana F, Voilley N, Lazdunski M. Molecular cloning and functional expression of a novel amiloride-sensitive Na+ channel. J Biol Chem. 1995 Nov. 17; 270(46):27411-4. PubMed PMID: 7499195. [0155] 75. Wang J, Yu T, Yin L, Li J, Yu L, Shen Y, Yu Y, Shen Y, Fu Q. Novel mutations in the SCNN1A gene causing Pseudohypoaldosteronism type 1. PLoS One. 2013 Jun. 6; 8(6):e65676. doi: 10.1371/journal.pone.0065676. Print 2013. PubMed PMID: 23762408; PubMed Central PMCID: PMC3675083. [0156] 76. Welzel M, Akin L, Bscher A, Gran T, Hauffa B P, Hgler W, Leonards J, Karges B, Kentrup H, Kirel B, Senses E E, Tekin N, Holterhus P M, Riepe F G. Five novel mutations in the SCNN1A gene causing autosomal recessive pseudohypoaldosteronism type 1. Eur J Endocrinol. 2013 Apr. 15; 168(5):707-15. doi: 10.1530/EJE-12-1000. Print 2013 May. PubMed PMID: 23416952. [0157] 77. Wong G P, Levine D. Congenital pseudohypoaldosteronism presenting in utero with acute polyhydramnios. J Matern Fetal Med. 1998 March-April; 7(2):76-8. PubMed PMID: 9584819. [0158] 78. Zennaro M C, Lombs M. Mineralocorticoid resistance. Trends Endocrinol Metab. 2004 August; 15(6):264-70. Review. PubMed PMID: 15358279.