Inhibition of ENaC activity by novel peptide trypsin-like inhibitors derived from amphibian skin secretions

E.L. Carroll, L.E.J. Douglas, J.A. Reihill, M. Zhou, T. Chen, L.P. McGarvey, F.T. Lundy, M.A. Hollywood, A. Crilly, J.C. Lockhart, S.L. Martin

Research output: Contribution to journalMeeting Abstract

Abstract

ENaC is activated by trypsin-like (TL) channel activating proteases
(CAPs), inhibition of which in cystic fibrosis has been shown to increase airways surface liquid and normalise mucociliary clearance (MCC)1. These proteases have potential to be therapeutic targets in other chronic airways diseases. This study investigated the ability of natural peptide TL inhibitors, derived from amphibian skin secretions, to inactivate ENaC via CAP inhibition.

Initial screening of the frog peptides was conducted using FRT cells stably transfected with ENaC and changes in fluorescence intensity (FLIPR) utilised to measure ENaC activity. Second round testing was performed using primary differentiated human airway epithelial cells (hAECs) by measurement of IEQ (equivalent short-circuit current) (TECC-24; EP Devices).

FLIPR revealed that all of the frog peptides significantly inhibited
ENaC, particularly QUB-1916 which elicited almost immediate,
complete channel inhibition. Different trends were observed when
the peptides were tested on hAECs using short-circuit current,
with other peptides demonstrating superior ENaC inhibition compared to QUB-1916. Initial profiling indicates a differential protease expression pattern between cell types which may underlie this discrepancy.

These findings highlight the importance of direct testing of channel activity using hAECs. Potential lead compounds are currently being evaluated for efficacy in airway hydration and MCC
studies.
Original languageEnglish
Pages (from-to)S250-S251
Number of pages2
JournalIrish Journal of Medical Science
Volume187
Issue numberSupplement 8
DOIs
Publication statusPublished - Aug 2018

Fingerprint

Trypsin Inhibitors
Amphibians
Peptide Hydrolases
Skin
Peptides
Epithelial Cells
Anura
Mucociliary Clearance
Human Activities
Cystic Fibrosis
Trypsin
Chronic Disease
Fluorescence
Equipment and Supplies

Keywords

  • COPD
  • BREATH
  • ENaC
  • interreg va
  • SEUPB
  • trypsin

Cite this

Carroll, E. L., Douglas, L. E. J., Reihill, J. A., Zhou, M., Chen, T., McGarvey, L. P., ... Martin, S. L. (2018). Inhibition of ENaC activity by novel peptide trypsin-like inhibitors derived from amphibian skin secretions. Irish Journal of Medical Science, 187(Supplement 8), S250-S251. https://doi.org/10.1007/s11845-018-1898-7
Carroll, E.L. ; Douglas, L.E.J. ; Reihill, J.A. ; Zhou, M. ; Chen, T. ; McGarvey, L.P. ; Lundy, F.T. ; Hollywood, M.A. ; Crilly, A. ; Lockhart, J.C. ; Martin, S.L. / Inhibition of ENaC activity by novel peptide trypsin-like inhibitors derived from amphibian skin secretions. In: Irish Journal of Medical Science. 2018 ; Vol. 187, No. Supplement 8. pp. S250-S251.
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title = "Inhibition of ENaC activity by novel peptide trypsin-like inhibitors derived from amphibian skin secretions",
abstract = "ENaC is activated by trypsin-like (TL) channel activating proteases(CAPs), inhibition of which in cystic fibrosis has been shown to increase airways surface liquid and normalise mucociliary clearance (MCC)1. These proteases have potential to be therapeutic targets in other chronic airways diseases. This study investigated the ability of natural peptide TL inhibitors, derived from amphibian skin secretions, to inactivate ENaC via CAP inhibition.Initial screening of the frog peptides was conducted using FRT cells stably transfected with ENaC and changes in fluorescence intensity (FLIPR) utilised to measure ENaC activity. Second round testing was performed using primary differentiated human airway epithelial cells (hAECs) by measurement of IEQ (equivalent short-circuit current) (TECC-24; EP Devices).FLIPR revealed that all of the frog peptides significantly inhibitedENaC, particularly QUB-1916 which elicited almost immediate,complete channel inhibition. Different trends were observed whenthe peptides were tested on hAECs using short-circuit current,with other peptides demonstrating superior ENaC inhibition compared to QUB-1916. Initial profiling indicates a differential protease expression pattern between cell types which may underlie this discrepancy.These findings highlight the importance of direct testing of channel activity using hAECs. Potential lead compounds are currently being evaluated for efficacy in airway hydration and MCCstudies.",
keywords = "COPD, BREATH, ENaC, interreg va, SEUPB, trypsin",
author = "E.L. Carroll and L.E.J. Douglas and J.A. Reihill and M. Zhou and T. Chen and L.P. McGarvey and F.T. Lundy and M.A. Hollywood and A. Crilly and J.C. Lockhart and S.L. Martin",
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Carroll, EL, Douglas, LEJ, Reihill, JA, Zhou, M, Chen, T, McGarvey, LP, Lundy, FT, Hollywood, MA, Crilly, A, Lockhart, JC & Martin, SL 2018, 'Inhibition of ENaC activity by novel peptide trypsin-like inhibitors derived from amphibian skin secretions', Irish Journal of Medical Science, vol. 187, no. Supplement 8, pp. S250-S251. https://doi.org/10.1007/s11845-018-1898-7

Inhibition of ENaC activity by novel peptide trypsin-like inhibitors derived from amphibian skin secretions. / Carroll, E.L.; Douglas, L.E.J.; Reihill, J.A.; Zhou, M.; Chen, T.; McGarvey, L.P.; Lundy, F.T.; Hollywood, M.A.; Crilly, A.; Lockhart, J.C.; Martin, S.L.

In: Irish Journal of Medical Science, Vol. 187, No. Supplement 8, 08.2018, p. S250-S251.

Research output: Contribution to journalMeeting Abstract

TY - JOUR

T1 - Inhibition of ENaC activity by novel peptide trypsin-like inhibitors derived from amphibian skin secretions

AU - Carroll, E.L.

AU - Douglas, L.E.J.

AU - Reihill, J.A.

AU - Zhou, M.

AU - Chen, T.

AU - McGarvey, L.P.

AU - Lundy, F.T.

AU - Hollywood, M.A.

AU - Crilly, A.

AU - Lockhart, J.C.

AU - Martin, S.L.

PY - 2018/8

Y1 - 2018/8

N2 - ENaC is activated by trypsin-like (TL) channel activating proteases(CAPs), inhibition of which in cystic fibrosis has been shown to increase airways surface liquid and normalise mucociliary clearance (MCC)1. These proteases have potential to be therapeutic targets in other chronic airways diseases. This study investigated the ability of natural peptide TL inhibitors, derived from amphibian skin secretions, to inactivate ENaC via CAP inhibition.Initial screening of the frog peptides was conducted using FRT cells stably transfected with ENaC and changes in fluorescence intensity (FLIPR) utilised to measure ENaC activity. Second round testing was performed using primary differentiated human airway epithelial cells (hAECs) by measurement of IEQ (equivalent short-circuit current) (TECC-24; EP Devices).FLIPR revealed that all of the frog peptides significantly inhibitedENaC, particularly QUB-1916 which elicited almost immediate,complete channel inhibition. Different trends were observed whenthe peptides were tested on hAECs using short-circuit current,with other peptides demonstrating superior ENaC inhibition compared to QUB-1916. Initial profiling indicates a differential protease expression pattern between cell types which may underlie this discrepancy.These findings highlight the importance of direct testing of channel activity using hAECs. Potential lead compounds are currently being evaluated for efficacy in airway hydration and MCCstudies.

AB - ENaC is activated by trypsin-like (TL) channel activating proteases(CAPs), inhibition of which in cystic fibrosis has been shown to increase airways surface liquid and normalise mucociliary clearance (MCC)1. These proteases have potential to be therapeutic targets in other chronic airways diseases. This study investigated the ability of natural peptide TL inhibitors, derived from amphibian skin secretions, to inactivate ENaC via CAP inhibition.Initial screening of the frog peptides was conducted using FRT cells stably transfected with ENaC and changes in fluorescence intensity (FLIPR) utilised to measure ENaC activity. Second round testing was performed using primary differentiated human airway epithelial cells (hAECs) by measurement of IEQ (equivalent short-circuit current) (TECC-24; EP Devices).FLIPR revealed that all of the frog peptides significantly inhibitedENaC, particularly QUB-1916 which elicited almost immediate,complete channel inhibition. Different trends were observed whenthe peptides were tested on hAECs using short-circuit current,with other peptides demonstrating superior ENaC inhibition compared to QUB-1916. Initial profiling indicates a differential protease expression pattern between cell types which may underlie this discrepancy.These findings highlight the importance of direct testing of channel activity using hAECs. Potential lead compounds are currently being evaluated for efficacy in airway hydration and MCCstudies.

KW - COPD

KW - BREATH

KW - ENaC

KW - interreg va

KW - SEUPB

KW - trypsin

U2 - 10.1007/s11845-018-1898-7

DO - 10.1007/s11845-018-1898-7

M3 - Meeting Abstract

VL - 187

SP - S250-S251

JO - Irish Journal of Medical Science

JF - Irish Journal of Medical Science

SN - 0021-1265

IS - Supplement 8

ER -