title: Esculentin-1a(1-21)NH2: a frog skin-derived peptide for microbial keratitis.
abstract: Pseudomonas aeruginosa is the primary bacterial pathogen causing contact lens related keratitis. Available ophthalmic agents have reduced efficacy and antimicrobial peptides (AMPs) hold promise as future antibiotics. Here we investigated the in vitro and in vivo anti-Pseudomonal activity of esculentin-1a(1-21)NH2, derived from a frog skin AMP. The data revealed a minimum inhibitory concentration between 2 and 16 μM against reference strains or drug-resistant clinical isolates of P. aeruginosa without showing toxicity to human corneal epithelial cells up to 50 μM. At 1 μM the peptide rapidly killed bacterial cells and this activity was fully retained in 150 mM sodium chloride and 70 % (v/v) human basal tears, particularly against the virulent ATCC 19660 strain. Furthermore, its dropwise administration at 40 μM to the ocular surface in a murine model of P. aeruginosa keratitis (three times daily, for 5 days post-infection) resulted in a significant reduction of infection. The mean clinical score was 2.89 ± 0.26 compared to 3.92 ± 0.08 for the vehicle control. In addition, the corneal level of viable bacteria in the peptide treated animals was significantly lower with a difference of 4 log10 colony counts, compared to 7.7 log10 cells recovered in the control. In parallel, recruitment of inflammatory cells was reduced by half compared to that found in the untreated eyes. Similar results were obtained when esculentin-1a(1-21)NH2 was applied prior to induction of keratitis. Overall, our findings highlight esculentin-1a(1-21)NH2 as an attractive candidate for the development of novel topical pharmaceuticals against Pseudomonas keratitis.
authors.affiliation: College of Optometry, The Ocular Surface Institute, University of Houston, Houston, TX, USA.
authors.forename: Satya Sree N
authors.initials: SSN
authors.lastname: Kolar
authors.affiliation: Department of Biochemical Sciences "A. Rossi Fanelli", Istituto Pasteur-Fondazione Cenci Bolognetti, Sapienza University of Rome, Via degli Apuli 9, 00185, Rome, Italy.
authors.forename: Vincenzo
authors.initials: V
authors.lastname: Luca
authors.affiliation: College of Optometry, The Ocular Surface Institute, University of Houston, Houston, TX, USA.
authors.forename: Hasna
authors.initials: H
authors.lastname: Baidouri
authors.affiliation: Ophthalmology Unit, NESMOS Department, S. Andrea Hospital, Faculty of Medicine and Psychology, Sapienza University of Rome, Rome, Italy.
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Mannino
authors.affiliation: College of Optometry, The Ocular Surface Institute, University of Houston, Houston, TX, USA.
authors.forename: Alison M
authors.initials: AM
authors.lastname: McDermott
authors.affiliation: Department of Biochemical Sciences "A. Rossi Fanelli", Istituto Pasteur-Fondazione Cenci Bolognetti, Sapienza University of Rome, Via degli Apuli 9, 00185, Rome, Italy. marialuisa.mangoni@uniroma1.it.
authors.forename: Maria Luisa
authors.initials: ML
authors.lastname: Mangoni
mesh_terms: D029845:Amphibian Proteins / Q000302:isolation & purification / Q000494:pharmacology*; D000704:Analysis of Variance; D000818:Animals; D023181:Antimicrobial Cationic Peptides / Q000302:isolation & purification / Q000494:pharmacology*; D001001:Anura / Q000378:metabolism*; D003261:Contact Lenses / Q000009:adverse effects / Q000382:microbiology*; D003315:Cornea / Q000166:cytology; D004847:Epithelial Cells / Q000187:drug effects; D006801:Humans; D007634:Keratitis / Q000188:drug therapy* / Q000209:etiology / Q000382:microbiology; D011550:Pseudomonas aeruginosa / Q000187:drug effects*; D012867:Skin / Q000737:chemistry*; D013778:Tetrazolium Salts; D013844:Thiazoles
chemical_list: D029845:Amphibian Proteins; D023181:Antimicrobial Cationic Peptides; D013778:Tetrazolium Salts; D013844:Thiazoles; C092874:esculentin protein, Rana esculenta; C022616:thiazolyl blue
references.citation: Curr Eye Res. 2007 Jul-Aug;32(7-8):595-609
references.pmid: 17852183
references.citation: Cornea. 1996 Jan;15(1):66-71
references.pmid: 8907383
references.citation: Invest Ophthalmol Vis Sci. 1995 May;36(6):1107-14
references.pmid: 7730020
references.citation: Cornea. 2008 Jan;27(1):22-7
references.pmid: 18245962
references.citation: Ophthalmology. 1999 Jul;106(7):1319-23
references.pmid: 10406614
references.citation: Invest Ophthalmol Vis Sci. 1993 Aug;34(9):2699-712
references.pmid: 8344792
references.citation: Trends Immunol. 2009 Mar;30(3):131-41
references.pmid: 19217824
references.citation: Nature. 2003 Nov 20;426(6964):306-10
references.pmid: 14628055
references.citation: Ann N Y Acad Sci. 2009 Apr;1163:75-82
references.pmid: 19456329
references.citation: ACS Appl Mater Interfaces. 2013 Dec 26;5(24):12789-93
references.pmid: 24266746
references.citation: Cornea. 2009 Sep;28(8):918-26
references.pmid: 19654521
references.citation: Arch Ophthalmol. 1986 Jan;104(1):79-83
references.pmid: 3942549
references.citation: Eye Contact Lens. 2003 Oct;29(4):207-9
references.pmid: 14555893
references.citation: Antimicrob Agents Chemother. 2007 Nov;51(11):3853-60
references.pmid: 17724155
references.citation: J Neurosci. 2006 Mar 1;26(9):2369-79
references.pmid: 16510714
references.citation: Cornea. 2012 Jul;31(7):777-9
references.pmid: 22378117
references.citation: Protein Pept Lett. 2010 Nov;17(11):1334-44
references.pmid: 20673228
references.citation: Sultan Qaboos Univ Med J. 2009 Aug;9(2):184-95
references.pmid: 21509299
references.citation: FEMS Microbiol Lett. 2002 Jan 10;206(2):143-9
references.pmid: 11814654
references.citation: Cell Mol Life Sci. 2013 Aug;70(15):2773-86
references.pmid: 23503622
references.citation: Semin Immunopathol. 2007 Apr;29(1):27-43
references.pmid: 17621952
references.citation: Cell Mol Life Sci. 2006 May;63(9):1060-9
references.pmid: 16572270
references.citation: Clin Exp Optom. 2011 Mar;94(2):161-8
references.pmid: 21083760
references.citation: Regul Pept. 2004 May 15;118(3):135-41
references.pmid: 15003829
references.citation: J Antimicrob Chemother. 2002 Jan;49(1):25-30
references.pmid: 11751763
references.citation: J Appl Microbiol. 2008 Dec;105(6):1817-25
references.pmid: 19016975
references.citation: J Clin Invest. 1998 Sep 1;102(5):874-80
references.pmid: 9727055
references.citation: Microbiol Mol Biol Rev. 2010 Dec;74(4):621-41
references.pmid: 21119020
references.citation: Ophthalmology. 2005 Dec;112(12):2172-9
references.pmid: 16325711
references.citation: Curr Eye Res. 2000 Mar;20(3):157-65
references.pmid: 10694889
references.citation: Br J Ophthalmol. 2006 May;90(5):609-11
references.pmid: 16622092
references.citation: Invest Ophthalmol Vis Sci. 2010 Jan;51(1):390-5
references.pmid: 19710414
references.citation: Invest Ophthalmol Vis Sci. 1996 Mar;37(4):534-43
references.pmid: 8595953
references.citation: Cell Mol Life Sci. 2011 Jul;68(13):2267-80
references.pmid: 21573781
references.citation: Biopolymers. 2002;66(4):236-48
references.pmid: 12491537
references.citation: Ophthalmology. 2006 Jan;113(1):109-16
references.pmid: 16360210
references.citation: Arch Ophthalmol. 1990 Jun;108(6):861-4
references.pmid: 2112378
references.citation: Eye Contact Lens. 2013 Jan;39(1):73-8
references.pmid: 23266587
references.citation: J Biol Chem. 1994 Apr 22;269(16):11956-61
references.pmid: 8163497
references.citation: Cornea. 1997 Jan;16(1):101-6
references.pmid: 8985641
references.citation: Exp Eye Res. 2013 Dec;117:53-61
references.pmid: 23880529
references.citation: J Pept Sci. 2009 Sep;15(9):607-14
references.pmid: 19507197
references.citation: Cornea. 1992 Sep;11(5):370-5
references.pmid: 1424662
references.citation: Int J Oral Sci. 2011 Apr;3(2):55-65
references.pmid: 21485309
references.citation: Cornea. 2008 Apr;27(3):320-6
references.pmid: 18362661
references.citation: Mol Biosyst. 2011 Mar;7(3):711-3
references.pmid: 21229178
references.citation: Invest Ophthalmol Vis Sci. 1987 Dec;28(12):1978-85
references.pmid: 3119512
references.citation: Nat Rev Drug Discov. 2011 Dec 16;11(1):37-51
references.pmid: 22173434
references.citation: Am J Ophthalmol. 2013 Jun;155(6):961-970.e2
references.pmid: 23601656
references.citation: Curr Opin Infect Dis. 2007 Apr;20(2):129-41
references.pmid: 17496570
references.citation: Invest Ophthalmol Vis Sci. 2003 May;44(5):1859-65
references.pmid: 12714616
references.citation: Curr Opin Immunol. 2006 Feb;18(1):24-30
references.pmid: 16337365
references.citation: Curr Drug Targets. 2009 Sep;10(9):895-905
references.pmid: 19799544
references.citation: Eye Contact Lens. 2013 Jan;39(1):67-72
references.pmid: 23266590
references.citation: Trends Microbiol. 2005 Jan;13(1):34-40
references.pmid: 15639630
references.citation: CLAO J. 1996 Apr;22(2):114-7
references.pmid: 8728617
references.citation: Nat Biotechnol. 2006 Dec;24(12):1551-7
references.pmid: 17160061
references.citation: FASEB J. 2001 Jun;15(8):1431-2
references.pmid: 11387247
references.citation: Clin Immunol. 2010 Apr;135(1):1-11
references.pmid: 20116332
references.citation: Invest Ophthalmol Vis Sci. 2013 Jan 07;54(1):175-82
references.pmid: 23211820
references.citation: Cell. 1997 Feb 21;88(4):553-60
references.pmid: 9038346
references.citation: Curr Eye Res. 2001 Jan;22(1):64-7
references.pmid: 11402380
references.citation: Curr Eye Res. 2005 Jul;30(7):535-42
references.pmid: 16020287
references.citation: Invest Ophthalmol Vis Sci. 2005 Feb;46(2):470-8
references.pmid: 15671271
references.citation: Science. 1999 May 21;284(5418):1318-22
references.pmid: 10334980
references.citation: Biopolymers. 2013 Nov;100(6):572-83
references.pmid: 23553602
references.citation: Nat Rev Immunol. 2003 Sep;3(9):710-20
references.pmid: 12949495
references.citation: Science. 1994 Apr 15;264(5157):359
references.pmid: 17836898
references.citation: Trans Am Ophthalmol Soc. 2002;100:243-71
references.pmid: 12545697