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Principal Investigator: DANIEL J GIBSON
Organization: INTEGUMED, LLC
Fiscal Year: 2021
Award: $299,749
Funding agency: National Institute of Diabetes and Digestive and Kidney Diseases
ABSTRACT
Chronic wounds affect over 2.5 million patients in the US alone, lasting on average 12-13 months and recurring
in 60-70% of patients. These occur primarily in lower extremities and vascular disease, diabetes, nephropathy,
and metabolic dysfunction are prevalent co-morbidities. These wounds frequently harbor bacterial infections as
mixed Gram-negative (e.g. P. aeruginosa, PA) and Gram-positive biofilms (e.g. S. aureus, SA), conferring
resistance to antibiotic therapy and inhibiting resolution. The heterogenous biofilm barrier, composed of lipids,
proteins, and polysaccharides, presents a robust physiochemical barrier and substrate for persistent infection.
Transition of planktonic bacteria to the distinct biofilm state requires a suitable substrate for specific adhesin
receptors, such as those found in host membranes and extracellular matrix (ECM), or nonspecific
physiochemical binding, enabling aggregation and biofilm ECM organization. This includes changes in
microbial lipid composition, secretion of lipid quorum signals and lipase virulence factors, and shift to oxidative
lipid metabolism to enable persistent colonization and high-density proliferation. Recent advances in ultra-high-
performance liquid chromatography, high resolution mass spectrometry, and bioinformatics technologies have
enabled detailed and accurate definition of the bacterial lipidome, aiding in identification of lipid pathways and
targets for antimicrobial activity using a systems biology approach.
Wound healing is further complicated by pro-inflammatory endogenous phospholipase activities. We propose
that lipid ether amines (LEA) represent a platform for novel topical wound treatments with phospholipid anti-
metabolite activity, counteracting both host and pathogen pathogenic mechanisms. As preliminary data, we
demonstrate >3-log (>1000x) colony forming unit reductions in multiple in vitro cultured, antibiotic resistant
biofilms and optimization of anti-metabolite activity. Phase I will synthesize additional candidates, screen for
biofilm reduction and activity in host cells, selecting a platform lead to determine in vivo wound healing effects
using biofilm infected db/db diabetic mouse model. LC-MS/MS analysis will be used to characterize bacterial or
host lipidomes and treatment effect in each Aim. In Aim I, LEA candidate compounds will be screened for
antimicrobial effect using clinically relevant, drug resistant ESKAPE (E. faecium, S. aureus, K. pneumoniae, A.
baumannii, P. aeruginosa, Enterobacter spp.) pathogens in cultured biofilms, including a translatable wounded
ex vivo pig skin explant model. In Aim II, LEA effects on migration and proliferation will be assessed using in
vitro full thickness skin construct. In Aim III, the lead compound will be applied in a diabetic mouse wound
healing model infected with PA or SA biofilms, to assess in vivo microbicidal and wound closure effects.
In Phase II, we will escalate in vivo model complexity by inoculating with mixed bacterial biofilms, refine
mechanism using in vitro binding and knock-down/out, evaluate potential adaptive resistance mechanisms,
conduct comprehensive ADME-T study, and begin in vivo large mammal Proof of Concept for IND submission.
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pathogens><Endotoxins><Enterobacter><Enterococcus faecium><Enzyme-Linked Immunosorbent Assay><Ethers><Exotoxins><Extracellular Matrix><Family suidae><Fluorescence><Formulation><Functional disorder><Generalized Growth><Genetic Alteration><Genetic Change><Genetic defect><Glycans><Goals><Growth><HPLC><Health><Health Care Systems><Healthcare><Healthcare Systems><High Performance Liquid Chromatography><High Pressure Liquid Chromatography><High Speed Liquid Chromatography><Histology><Human><Humulin R><Image><Impaired tissue repair><Impaired wound healing><In Situ><In Vitro><Inflammation><Inflammatory><Insulin><Intracellular Communication and Signaling><K pneumoniae><K. pneumoniae><Keratin><Kidney Diseases><Klebsiella pneumoniae><Lead><Lecithinases><Ligands><Lipase><Lipids><Lower Extremity><Lower Limb><Machine Learning><Mammalia><Mammals><Mass Photometry/Spectrum Analysis><Mass Spectrometry><Mass Spectroscopy><Mass Spectrum><Mass Spectrum Analyses><Mass Spectrum Analysis><Membrane><Membrum inferius><Metabolic dysfunction><Mice><Mice Mammals><Microbial Biofilms><Modeling><Modern Man><Modification><Molecular Interaction><Murine><Mus><Mutation><Nephropathy><Novolin R><Obesity><Outcome Measure><P aeruginosa><P. aeruginosa><P. aeruginosa infection><Pathogenicity><Pathogenicity Factors><Pathway interactions><Patients><Pb element><Phase><Phosphatides><Phospholipase><Phospholipids><Physiopathology><Pigs><Plasma><Plasma Membrane><Plasma Serum><Plasmids><Polysaccharides><Property><Proteins><Pseudomonas aeruginosa><Pseudomonas aeruginosa infection><Pseudomonas pyocyanea><Punch Biopsy><Receptor Protein><Regular Insulin><Renal Disease><Resistance><Resistance to antibiotics><Resistant to antibiotics><Resolution><Reticuloendothelial System, Serum, Plasma><S aureus><S faceium><S faecium><S. aureus><S. faceium><S. faecium><Safety><Sampling><Signal Transduction><Signal Transduction Systems><Signaling><Skin><Skin wound healing><Staining method><Stains><Staph aureus><Staphylococcus aureus><Sterile coverings><Streptococcus faceium><Streptococcus faecium><Suidae><Supervision><Swine><Systems Biology><Technology><Testing><Therapeutic><Therapeutic Effect><Thick><Thickness><Tissue Growth><Topical Drug Administration><Topical application><Toxic effect><Toxicities><Triacylglycerol Hydrolase><Triacylglycerol Lipase><Triacylglycerol acylhydrolase><Tributyrinase><Triglyceridase><Triglyceride Lipase><Triolean Hydrolase><Universities><Vascular Diseases><Vascular Disorder><Virulence><Virulence Factors><Wound Repair><Wound models><abnormal tissue repair><adhesin><adiposity><administer topically><amine><amphiphilicity><analog><anti-microbial><anti-microbial effect><antibiotic drug resistance><antibiotic resistant><antibiotic resistant bacteria><antimicrobial><apply topically><bacteria infection><bacteria metabolism><bacterial antibiotic resistant><bacterial disease><bacterial disease treatment><bacterial infectious disease treatment><bacterial metabolism><bacterial resistance to 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