Breaking the Barrier: Discovery of Antibiotic Resistance Factors and Novel Ocean Drugs Peter D. R. Moeller, Ph.D. Toxin/Natural Products Chemistry National Ocean Service/NOAA Hollings Marine Laboratory Charleston, SC

Agelas conifera Diseased coral reef – image taken from A Sponge happily survives where everything else is dying. How? How? This Sponge provided us with a clue from nature on how to deal with pathogenic microbes!

Survival Against Bacterial Infection Dr. Ernesto Weil, University of Puerto Rico- Mayaquez, PR Marine organisms dwell in a microbial-rich environment Protect themselves against biofouling through the production of anti-biofilm natural compounds Utilize antibiotics to deter attack from infectious microbes These mechanisms can be used in a 1-2 punch approach to drug discovery

1 st Punch: Production of Anti-Biofilm Chemicals Bacteria often live as a community encased in a protective extracellular matrix (biofilm) attached to a biotic or abiotic surface. These biofilms protect bacteria effectively against coming into contact with antibiotics Removal of this protective biofilm-encasing may allow conventional antibiotics to work against multi-drug resistant bacteria

Biofilm Relevance To Human Health Bacteria in biofilms are 1,000-fold more resistant to antibiotics 65-80% of all bacterial infections are biofilm-based Major cause of mortality in CF patients Cause infection of indwelling medical devices Nosocomial infections Destruction of citrus and other agricultural products Corrosion of petroleum pipelines

Agelas conifera Diseased coral reef – image taken from Novel chemicals designed for biofilm inhibition/elimination Chemical Defense Factory – Targeted, Selective, Efficient

Cis compound (CAGE)Trans compound (TAGE) Biofilms of Pseudomonas aeruginosa One of the causes of mortality in cystic fibrosis (CF) patients Derivatives of Sponge-Derived Ageliferin Biofilm Disrupting Agents

Anti-biofilm Chemicals Enable Effective Use of Current Generation Antibiotics A. baumannii Control AntibioticAnti- biofilm agent Antibiotic + Antibiofilm agent Acinetobacter baumannii Isolated from military personnel wounded in Afghanistan MRSA Methicillin Resistant Staphylococcus aureus MRSA Control Methicillin Antibiotic Anti- biofilm agent Antibiotic + Antibiofilm agent

Pseudomonas aeruginosa: Opportunistic pathogen infecting wounded soldiers, and indwelling medical devices. Significant source of mortality for cancer patients and CF patients. Acinetobacter baumannii: Opportunistic pathogen infecting wounded soldiers and indwelling medical devices. Responsible for hospital closures and death of wounded war fighters. Multi- drug resistant strains to every known antibiotic have been isolated. Vibrio vulnificus: Opportunistic pathogen that causes septicemia and seafood poisoning. It is related to the bacteria that causes Cholera. Haemophilus influenzae: Opportunistic pathogen responsible for ear infections. Bordetella: Pathogen that causes whooping cough. Responsible for 300,000 deaths/year. Staphylococcus aureus and epidermidis: Bacterial pathogens that are the prevalently isolated from nosocomial infections. Has risen to prominence due to MRSA. Halomonas pacifica: Biofouling of ships. Efficacy Demonstrated Against Significant Infectious Bacteria

2 nd Punch: Antibiotic Discovery A Focus on Pseudopterogorgia americana A Caribbean Coral Media Luna reef, La Parguera, Puerto Rico The Caribbean has lost over 80% of its hard coral cover in the past years. Visually healthy P. americana Visually diseased P. americana (close-up) Image Credit: Dr. Ernesto Weil, University of Puerto Rico- Mayaquez, PR

Chemical Warfare: Antimicrobial Screening Assay Coral Isolate A vs K. rhizophila Coral Isolate B vs P. aeruginosa Coral Pathogens: Vibrio coralliilyticus Aspergillus sydowii Human Pathogens: Pseudomonas aeruginosa Escherichia coli Kocuria rhizophila Bacillus spp. (B. subtilis) Aspergillus spp. (A. sydowii) Marine Invertebrate Pathogen: V. harveyi Inhibition Bacterial isolates tested against a series of pathogenic bacteria Antimicrobial screening often highlights potential antibiotic selectivity Selective antibiotic activity targets potential for “personalized medicine”

Isolating Novel Antibiotics from a Marine Bacterium Pseudovibrio denitrificans Recently discovered Genus specific to the marine environment Demonstrated strong broad spectrum antibiotic activity Produces several ‘suites’ of antibiotics, some with high selectivity. Image Credit: Ben Neely, MUSC 1 Sertan-de Guzman et al., 2007; 2 Toledo et al., 2006; 3 Hosoya and Yokota 2007 “Bring it on” species with attitude! Loaded for Bear

Microbial Ecology: Microbe-microbe interactions Stressors and Response: Symbiont-pathogen interaction Chemical Ecology The study of chemicals involved in the interactions of organisms within an ecosystem, with a focus on production and response to natural products Chemical Defense: Bioactive compound production Marine Natural Products Chemistry Anti-cancer, Anti-fungal, Antibiotics, New Health Related Productions of Interest, New Agricultural Niche Crops screening for viability

Acknowledgements Christian Melander, Ph.D. Assistant Professor Department of Chemistry North Carolina State University John Cavanagh, Ph.D. William Neal Reynolds Distinguished Professor Department of Molecular & Structural Biochemistry North Carolina State University Anti-Biofilm ResearchAntibiotic Discovery Pamela J. Morris, Ph.D. Associate Professor Cell Biology and Anatomy Marine Biomedicine and Environmental Sciences Center Medical University of South Carolina Hollings Marine Laboratory Maria Vizcaino Ph.D. Candidate Marine Biomedicine and Environmental Sciences Center Medical University of South Carolina Hollings Marine Laboratory Funding Sources NSF, NIH, SC SEAGRANT, NOS/NOAA