Disparate Regions of the Human Chemokine CXCL10 Exhibit Broad-Spectrum Antimicrobial Activity against Biodefense and Antibiotic-Resistant Bacterial Pathogens

Crawford, Matthew A.; Ward, Amanda E.; Gray, Vincent P.; Bailer, P; Fisher, Debra J.; Kubicka, Ewa; Cui, Zixian; Luo, Qinmo; Gray, Mary C.; Criss, Alison K.; Lum, Lawrence G.; Tamm, Lukas K.; Letteri, Rachel A.; Hughes, Molly A.

doi:10.1021/acsinfecdis.2c00456

Cited by 6 publications

(10 citation statements)

References 71 publications

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“…For the AMP component of our conjugates in this study, we selected a previously studied antimicrobial peptide, called P9, which is derived from the C-terminus of the human chemokine CXCL10. , To help retain the helical structure of this chemokine-derived peptide, a hydrocarbon staple and N-terminal acetyl (Ac) cap are installed . The resulting peptide, stapled Ac-P9, is active against a range of antibiotic-resistant bacterial isolates and does not exhibit significant hemolysis at concentrations commensurate with bacterial killing.…”

Section: Resultsmentioning

confidence: 99%

“…The antimicrobial activity of peptides and conjugates was evaluated against a well-characterized multidrug-resistant (MDR) clinical isolate, Klebsiella pneumoniae BL13802, ,, using the metabolic indicator dye alamarBlue to quantify bacterial viability/survival, as previously described. ,− Briefly, logarithmic-phase bacteria (grown in LB medium to an OD 600 of ∼0.6) were diluted in RPMI medium and combined in a 1:1 ratio with medium alone or containing peptides/conjugates in the wells of a 96-well plate (200 μL total volume with ∼2.5 × 10 5 CFU/mL and final peptide equivalent concentrations of 200 or 100 μM). Wells containing only RPMI medium were included in each experiment as a sample blank.…”

Section: Methodsmentioning

confidence: 99%

“…Circular dichroism spectroscopy was run under N 2(g) in a 0.1 cm path length quartz cell at 25 °C using a JASCO (Easton, Maryland) J-1500 CD spectrophotometer with a Peltier thermostated single-position cell holder. Peptide or conjugate samples were prepared at 0.1 mg/mL in 10 mM phosphate buffer (PB, pH = 7) or in 50% v/v TFE or 60 mM SDS to mimic the hydrophobic environment of anionic bacterial cell membranes. − Spectra were obtained in triplicate on the same solution from 180 to 250 nm with a scanning speed of 50 nm/min and an integration time of 1 s. Mean residue ellipticity was calculated as ellipticity (θ, mdeg)/[10 × path length ( l , cm) × peptide concentration ( C , mol/L) × number of amino acid residues] (Figure S9). Conjugate concentrations were calculated based on the average molecular weight determined from NMR spectra, inclusive of TFA counterions on each cationic Lys and Arg residues, as well as protons on each anionic Glu residue.…”

Section: Methodsmentioning

confidence: 99%

“…33,42 To help retain the helical structure of this chemokine-derived peptide, a hydrocarbon staple and Nterminal acetyl (Ac) cap are installed. 33 The resulting peptide, stapled Ac-P9, is active against a range of antibiotic-resistant bacterial isolates and does not exhibit significant hemolysis at concentrations commensurate with bacterial killing. As this helical peptide resembles many traditional membrane-disrupting AMPs in its amphipathic character, 5 we envision that the design rules established here will be useful to conceiving other useful AMP−polymer conjugates.…”

Section: Synthesis Of Stapled Ac-cggp9-peg Conjugatesmentioning

confidence: 99%

“…Normalization of this signal, relative to that produced in untreated bacteria, yields a percentage of surviving bacteria (Figure 4a). 33 Similar to our solution property studies, we held peptide concentrations constant to show how arm number and arm length affect antimicrobial activity. We tested two peptide equivalent concentrations (i.e., 100 and 200 μM) that highlight differences among the conjugates (Figure S15).…”

Section: Bactericidal Activity Against K Pneumoniaementioning

confidence: 99%

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Antimicrobial Peptide–Poly(ethylene glycol) Conjugates: Connecting Molecular Architecture, Solution Properties, and Functional Performance

Cui,

Crawford,

Rumble

et al. 2023

ACS Polym. Au

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Antimicrobial peptides (AMPs) are promising alternatives to conventional antibiotics for treating infections caused by drug-resistant bacteria; yet, many peptides are limited by toxicity to eukaryotic cells and instability in biological environments. Conjugation to linear polymers that reduce cytotoxicity and improve stability, however, often decreases antimicrobial activity. In this work, we combine the biocompatibility advantages of poly(ethylene glycol) (PEG) with the efficacy merits of nonlinear polymer architectures that accommodate multiple AMPs per molecule. By conjugating a chemokine-derived AMP, stapled Ac-P9, to linear and star-shaped PEG with various arm numbers and lengths, we investigated the role of molecular architecture in solution properties (i.e., ζ-potential, size, and morphology) and performance (i.e., antimicrobial activity, hemolysis, and protease resistance). Linear, 4-arm, and 8-arm conjugates with 2−2.5 kDa PEG arms were found to form nanoscale structures in solution with lower ζ-potentials relative to the unconjugated AMP, suggesting that the polymer partially shields the cationic AMP. Reducing the length of the PEG arms of the 8-arm conjugate to 1.25 kDa appeared to better reveal the peptide, seen by the increased ζ-potential, and promote assembly into particles with a larger size and defined spherical morphology. The antimicrobial effects exerted by the short 8-arm conjugate rivaled that of the unconjugated peptide, and the AMP constituents of the short 8-arm conjugate were protected from proteolytic degradation. All other conjugates examined also imparted a degree of protease resistance, but exhibited some reduced level of antimicrobial activity as compared to the AMP alone. None of the conjugates caused significant cytotoxic effects, which bodes well for their future potential to treat infections. While enhancing proteolytic stability often comes with the cost of lower antimicrobial activity, we have found that presenting AMPs at high density on a neutral nonlinear polymer strikes a favorable balance, exhibiting both enhanced stability and high antimicrobial activity.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Synthesis Of Stapled Ac-cggp9-peg Conjugatesmentioning

confidence: 99%

Section: Bactericidal Activity Against K Pneumoniaementioning

confidence: 99%

See 3 more Smart Citations

Antimicrobial Peptide–Poly(ethylene glycol) Conjugates: Connecting Molecular Architecture, Solution Properties, and Functional Performance

Cui,

Crawford,

Rumble

et al. 2023

ACS Polym. Au

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Inflammatory liver diseases and susceptibility to sepsis

2024

Clinical Science

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Patients with inflammatory liver diseases, particularly alcohol-associated liver disease and metabolic dysfunction-associated fatty liver disease (MAFLD), have higher incidence of infections and mortality rate due to sepsis. The current focus in the development of drugs for MAFLD is the resolution of non-alcoholic steatohepatitis and prevention of progression to cirrhosis. In patients with cirrhosis or alcoholic hepatitis, sepsis is a major cause of death. As the metabolic center and a key immune tissue, liver is the guardian, modifier, and target of sepsis. Septic patients with liver dysfunction have the highest mortality rate compared with other organ dysfunctions. In addition to maintaining metabolic homeostasis, the liver produces and secretes hepatokines and acute phase proteins (APPs) essential in tissue protection, immunomodulation, and coagulation. Inflammatory liver diseases cause profound metabolic disorder and impairment of energy metabolism, liver regeneration, and production/secretion of APPs and hepatokines. Herein, the author reviews the roles of (1) disorders in the metabolism of glucose, fatty acids, ketone bodies, and amino acids as well as the clearance of ammonia and lactate in the pathogenesis of inflammatory liver diseases and sepsis; (2) cytokines/chemokines in inflammatory liver diseases and sepsis; (3) APPs and hepatokines in the protection against tissue injury and infections; and (4) major nuclear receptors/signaling pathways underlying the metabolic disorders and tissue injuries as well as the major drug targets for inflammatory liver diseases and sepsis. Approaches that focus on the liver dysfunction and regeneration will not only treat inflammatory liver diseases but also prevent the development of severe infections and sepsis.

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Nasal microbionts differentially colonize and elicit cytokines in human nasal epithelial organoids

Boyd,

Kafer,

Escapa

et al. 2024

Preprint

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Nasal colonization byStaphylococcus aureusorStreptococcus pneumoniaeis associated with an increased risk of infection by these pathobionts, whereas nasal colonization byDolosigranulumspecies is associated with health. Human nasal epithelial organoids (HNOs) physiologically recapitulate human nasal respiratory epithelium with a robust mucociliary blanket. We reproducibly monocolonized HNOs with these three bacteria for up to 48 hours with varying kinetics across species. HNOs tolerated bacterial monocolonization with localization of bacteria to the mucus layer and minimal cytotoxicity compared to uncolonized HNOs. Human nasal epithelium exhibited both species-specific and general cytokine responses, without induction of type I interferons, consistent with colonization rather than infection. Only liveS. aureuscolonization induced IL-1 family cytokines, suggestive of inflammasome signaling.D. pigrumand liveS. aureusdecreased CXCL10, whereasS. pneumoniaeincreased CXCL11, chemokines involved in antimicrobial responses. HNOs are a compelling model system to reveal host-microbe dynamics at the human nasal mucosa.

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Disparate Regions of the Human Chemokine CXCL10 Exhibit Broad-Spectrum Antimicrobial Activity against Biodefense and Antibiotic-Resistant Bacterial Pathogens

Cited by 6 publications

References 71 publications

Antimicrobial Peptide–Poly(ethylene glycol) Conjugates: Connecting Molecular Architecture, Solution Properties, and Functional Performance

Antimicrobial Peptide–Poly(ethylene glycol) Conjugates: Connecting Molecular Architecture, Solution Properties, and Functional Performance

Inflammatory liver diseases and susceptibility to sepsis

Nasal microbionts differentially colonize and elicit cytokines in human nasal epithelial organoids

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