Ancient Antimicrobial Peptides Kill Antibiotic-Resistant Pathogens: Australian Mammals Provide New Options

Background To overcome the increasing resistance of pathogens to existing antibiotics the 10×'20 Initiative declared the urgent need for a global commitment to develop 10 new antimicrobial drugs by the year 2020. Naturally occurring animal antibiotics are an obvious place to start. The recently sequenced genomes of mammals that are divergent from human and mouse, including the tammar wallaby and the platypus, provide an opportunity to discover novel antimicrobials. Marsupials and monotremes are ideal potential sources of new antimicrobials because they give birth to underdeveloped immunologically naïve young that develop outside the sterile confines of a uterus in harsh pathogen-laden environments. While their adaptive immune system develops innate immune factors produced either by the mother or by the young must play a key role in protecting the immune-compromised young. In this study we focus on the cathelicidins, a key family of antimicrobial peptide genes. Principal Finding We identified 14 cathelicidin genes in the tammar wallaby genome and 8 in the platypus genome. The tammar genes were expressed in the mammary gland during early lactation before the adaptive immune system of the young develops, as well as in the skin of the pouch young. Both platypus and tammar peptides were effective in killing a broad range of bacterial pathogens. One potent peptide, expressed in the early stages of tammar lactation, effectively killed multidrug-resistant clinical isolates of Pseudomonas aeruginosa, Klebsiella pneumoniae and Acinetobacter baumannii. Conclusions and Significance Marsupial and monotreme young are protected by antimicrobial peptides that are potent, broad spectrum and salt resistant. The genomes of our distant relatives may hold the key for the development of novel drugs to combat multidrug-resistant pathogens.

[1]  M. Ågren,et al.  Stability of the cathelicidin peptide LL-37 in a non-healing wound environment. , 2011, Acta dermato-venereologica.

[2]  Yoshiyuki Sakaki,et al.  Genome sequence of an Australian kangaroo, Macropus eugenii, provides insight into the evolution of mammalian reproduction and development , 2011, Genome Biology.

[3]  Jing Wang,et al.  The antibacterial activity of BF-30 in vitro and in infected burned rats is through interference with cytoplasmic membrane integrity , 2011, Peptides.

[4]  P. Moore,et al.  Expression and Activity of a Novel Cathelicidin from Domestic Cats , 2011, PloS one.

[5]  Neang S. Ly,et al.  Resurgence of Colistin: A Review of Resistance, Toxicity, Pharmacodynamics, and Dosing , 2010, Pharmacotherapy.

[6]  Yun Zhang,et al.  Structure–function relationship of king cobra cathelicidin , 2010, Peptides.

[7]  M. V. van Hoek,et al.  Antimicrobial activity of the Naja atra cathelicidin and related small peptides. , 2010, Biochemical and biophysical research communications.

[8]  N. A. Jacques,et al.  An observational study of the microbiome of the maternal pouch and saliva of the tammar wallaby, Macropus eugenii, and of the gastrointestinal tract of the pouch young. , 2010, Microbiology.

[9]  A. Papenfuss,et al.  No evidence of expression of two classes of natural antibiotics (cathelicidins and defensins) in a sample of platypus milk , 2009 .

[10]  R. Gallo,et al.  AMPed up immunity: how antimicrobial peptides have multiple roles in immune defense. , 2009, Trends in immunology.

[11]  M. Baker,et al.  Identification and expression of a novel marsupial cathelicidin from the tammar wallaby (Macropus eugenii). , 2009, Veterinary immunology and immunopathology.

[12]  T. Koehler,et al.  Cathelicidin Administration Protects Mice from Bacillus anthracis Spore Challenge1 , 2008, The Journal of Immunology.

[13]  Yun Zhang,et al.  Identification and characterization of novel reptile cathelicidins from elapid snakes , 2008, Peptides.

[14]  Hailong Yang,et al.  Snake Cathelicidin from Bungarus fasciatus Is a Potent Peptide Antibiotics , 2008, PloS one.

[15]  J. Turnidge,et al.  In vitro pharmacodynamics of colistin against multidrug-resistant Klebsiella pneumoniae. , 2008, The Journal of antimicrobial chemotherapy.

[16]  Kerry A. Daly,et al.  Identification, characterization and expression of cathelicidin in the pouch young of tammar wallaby (Macropus eugenii). , 2008, Comparative biochemistry and physiology. Part B, Biochemistry & molecular biology.

[17]  Zhe‐Xi Luo,et al.  Transformation and diversification in early mammal evolution , 2007, Nature.

[18]  M. Cooper,et al.  Chicken cathelicidin-B1, an antimicrobial guardian at the mucosal M cell gateway , 2007, Proceedings of the National Academy of Sciences.

[19]  M. Nei,et al.  MEGA4: Molecular Evolutionary Genetics Analysis (MEGA) software version 4.0. , 2007, Molecular biology and evolution.

[20]  Ziheng Yang.  PAML 4: phylogenetic analysis by maximum likelihood. , 2007, Molecular biology and evolution.

[21]  Kate E. Jones,et al.  The delayed rise of present-day mammals , 1990, Nature.

[22]  R. Hancock,et al.  Antimicrobial and host-defense peptides as new anti-infective therapeutic strategies , 2006, Nature Biotechnology.

[23]  J. Soulages,et al.  Structure–activity relationships of fowlicidin‐1, a cathelicidin antimicrobial peptide in chicken , 2006, The FEBS journal.

[24]  M. Rocchi,et al.  LL-37 Protects Rats against Lethal Sepsis Caused by Gram-Negative Bacteria , 2006, Antimicrobial Agents and Chemotherapy.

[25]  S. Fernando,et al.  Identification and Functional Characterization of Three Chicken Cathelicidins with Potent Antimicrobial Activity* , 2006, Journal of Biological Chemistry.

[26]  R. Hancock,et al.  Cationic host defense (antimicrobial) peptides. , 2006, Current opinion in immunology.

[27]  Yongan Zhang,et al.  Two Cathelicidin Genes Are Present in both Rainbow Trout (Oncorhynchus mykiss) and Atlantic Salmon (Salmo salar) , 2006, Antimicrobial Agents and Chemotherapy.

[28]  F. Tödtling,et al.  One size fits all?: Towards a differentiated regional innovation policy approach , 2005 .

[29]  Yongan Zhang,et al.  Identification of a Novel Cathelicidin Gene in the Rainbow Trout, Oncorhynchus mykiss , 2005, Infection and Immunity.

[30]  M. Zanetti.  The role of cathelicidins in the innate host defenses of mammals. , 2005, Current issues in molecular biology.

[31]  David Posada,et al.  ProtTest: selection of best-fit models of protein evolution , 2005, Bioinform..

[32]  M. Zanetti,et al.  The cathelicidins--structure, function and evolution. , 2005, Current protein & peptide science.

[33]  N. Grishin,et al.  Reconstruction of ancestral protein sequences and its applications , 2004, BMC Evolutionary Biology.

[34]  Richard J. Edwards,et al.  GASP: Gapped Ancestral Sequence Prediction for proteins , 2004, BMC Bioinformatics.

[35]  M. Zasloff,et al.  Hagfish intestinal antimicrobial peptides are ancient cathelicidins , 2003, Peptides.

[36]  E. Isogai,et al.  Effect of human cationic antimicrobial protein 18 Peptide on endotoxin-induced uveitis in rats. , 2003, Investigative ophthalmology & visual science.

[37]  J. Old,et al.  The detection of mature T‐ and B‐cells during development of the lymphoid tissues of the tammar wallaby (Macropus eugenii) , 2003, Journal of anatomy.

[38]  James M. Wilson,et al.  Cathelicidins - a family of multifunctional antimicrobial peptides , 2003, Cellular and Molecular Life Sciences CMLS.

[39]  J. Old,et al.  Immunohistochemistry of the lymphoid tissues of the tammar wallaby, Macropus eugenii , 2002, Journal of anatomy.

[40]  S. Jackson,et al.  Reproductive behaviour and food consumption associated with the captive breeding of platypus (Ornithorhynchus anatinus) , 2002 .

[41]  J. Turnidge,et al.  In Vitro Pharmacodynamic Properties of Colistin and Colistin Methanesulfonate against Pseudomonas aeruginosaIsolates from Patients with Cystic Fibrosis , 2001, Antimicrobial Agents and Chemotherapy.

[42]  J. Old,et al.  Development of the immune system and immunological protection in marsupial pouch young. , 2000, Developmental and comparative immunology.

[43]  V. Solovyev,et al.  Ab initio gene finding in Drosophila genomic DNA. , 2000, Genome research.

[44]  J. Connolly,et al.  Histological and immunohistological investigation of lymphoid tissue in the platypus (Ornithorhynchus anatinus) , 1999, Journal of anatomy.

[45]  R. Bals,et al.  The peptide antibiotic LL-37/hCAP-18 is expressed in epithelia of the human lung where it has broad antimicrobial activity at the airway surface. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[46]  C. Tyndale-Biscoe,et al.  The Developing Marsupial: Models for Biomedical Research , 1988 .

[47]  G. Anderson,et al.  Methods in Mammalian Reproduction , 1978 .

[48]  G. Sharman.  Reproductive physiology of marsupials. , 1970, Science.

[49]  Sudhir Kumar,et al.  The timetree of life , 2009 .

[50]  Miriam K. Konkel,et al.  Tumour invasion and metastasis initiated by microRNA-10b in breast cancer , 2008, Nature.

[51]  F. Blecha,et al.  Canine cathelicidin (K9CATH): gene cloning, expression, and biochemical activity of a novel pro-myeloid antimicrobial peptide. , 2007, Developmental and comparative immunology.

[52]  T. Ganz,et al.  Inhibition of neutrophil elastase prevents cathelicidin activation and impairs clearance of bacteria from wounds. , 2001, Blood.

[53]  S Rozen,et al.  Primer3 on the WWW for general users and for biologist programmers. , 2000, Methods in molecular biology.

[54]  Sean R. Eddy,et al.  Profile hidden Markov models , 1998, Bioinform..

[55]  R. Lehrer,et al.  Designer assays for antimicrobial peptides. Disputing the "one-size-fits-all" theory. , 1997, Methods in molecular biology.

[56]  R. Lehrer,et al.  Designer Assays for Antimicrobial Peptides , 1997 .

[57]  Gapped BLAST and PSI-BLAST: A new , 1997 .

[58]  T. Grant,et al.  The platypus : a unique mammal , 1995 .

[59]  G. Grigg,et al.  Kangaroos, wallabies and rat-kangaroos , 1989 .

[60]  B. Green,et al.  The Composition of Marsupial Milk , 1988 .

[61]  M. Renfree,et al.  15 – Manipulation of Marsupial Embryos and Pouch Young , 1978 .