skip to main content


Title: Fusogenic porous silicon nanoparticles as a broad-spectrum immunotherapy against bacterial infections
Bacterial infections are re-emerging as substantial threats to global health due to the limited selection of antibiotics that are capable of overcoming antibiotic-resistant strains. By deterring such mutations whilst minimizing the need to develop new pathogen-specific antibiotics, immunotherapy offers a broad-spectrum therapeutic solution against bacterial infections. In particular, pathology resulting from excessive immune response ( i.e. fibrosis, necrosis, exudation, breath impediment) contributes significantly to negative disease outcome. Herein, we present a nanoparticle that is targeted to activated macrophages and loaded with siRNA against the Irf5 gene. This formulation is able to induce >80% gene silencing in activated macrophages in vivo , and it inhibits the excessive inflammatory response, generating a significantly improved therapeutic outcome in mouse models of bacterial infection. The versatility of the approach is demonstrated using mice with antibiotic-resistant Gram-positive (methicillin-resistant Staphylococcus aureus ) and Gram-negative ( Pseudomonas aeruginosa ) muscle and lung infections, respectively. Effective depletion of the Irf5 gene in macrophages is found to significantly improve the therapeutic outcome of infected mice, regardless of the bacteria strain and type.  more » « less
Award ID(s):
2011924 1603177
NSF-PAR ID:
10225984
Author(s) / Creator(s):
; ; ; ; ;
Date Published:
Journal Name:
Nanoscale Horizons
Volume:
6
Issue:
4
ISSN:
2055-6756
Page Range / eLocation ID:
330 to 340
Format(s):
Medium: X
Sponsoring Org:
National Science Foundation
More Like this
  1. The threat of antibiotic resistance warrants the discovery of agents with novel antimicrobial mechanisms. Antimicrobial peptides (AMPs) directly disrupting bacterial membranes may overcome resistance to traditional antibiotics. AMP development for clinical use has been mostly limited to topical application to date. We developed a rational framework for systematically addressing this challenge using libraries composed of 86 novel Trp- and Arg-rich engineered peptides tested against clinical strains of the most common multidrug-resistant bacteria known as ESKAPE pathogens. Structure-function correlations revealed minimum lengths (as low as 16 residues) and Trp positioning for maximum antibacterial potency with mean minimum inhibitory concentration (MIC) of 2–4 μM and corresponding negligible toxicity to mammalian cells. Twelve peptides were selected based on broad-spectrum activity against both gram-negative and -positive bacteria and <25% toxicity to mammalian cells at maximum test concentrations. Most of the selected PAX remained active against the colistin-resistant clinical strains. Of the selected peptides, the shortest (the 16-residue E35) was further investigated for antibacterial mechanism and proof-of-concept in vivo efficacy. E35 killed an extensively-resistant isolate of Pseudomonas aeruginosa (PA239 from the CDC, also resistant to colistin) by irreversibly disrupting the cell membranes as shown by propidium iodide incorporation, using flow cytometry and live cell imaging. As proof of concept, in vivo toxicity studies showed that mice tolerated a systemic dose of up to 30 mg/kg peptide and were protected with a single 5 mg/kg intravenous (IV) dose against an otherwise lethal intraperitoneal injection of PA239. Efficacy was also demonstrated in an immune-compromised Klebsiella pneumoniae infection model using a daily dose of 4mg/kg E35 systemically for 2 days. This framework defines the determinants of efficacy of helical AMPs composed of only cationic and hydrophobic amino acids and provides a path for a potential departure from the restriction to topical use of AMPs toward systemic application. 
    more » « less
  2. Background. Rapid blood culture diagnostics are of unclear benefit for patients with gram-negative bacilli (GNB) bloodstream infections (BSIs). We conducted a multicenter, randomized, controlled trial comparing outcomes of patients with GNB BSIs who had blood culture testing with standard-of-care (SOC) culture and antimicrobial susceptibility testing (AST) vs rapid organism identification (ID) and phenotypic AST using the Accelerate Pheno System (RAPID). Methods. Patients with positive blood cultures with Gram stains showing GNB were randomized to SOC testing with antimicrobial stewardship (AS) review or RAPID with AS. The primary outcome was time to first antibiotic modification within 72 hours of randomization. Results. Of 500 randomized patients, 448 were included (226 SOC, 222 RAPID). Mean (standard deviation) time to results was faster for RAPID than SOC for organism ID (2.7 [1.2] vs 11.7 [10.5] hours; P < .001) and AST (13.5 [56] vs 44.9 [12.1] hours; P < .001). Median (interquartile range [IQR]) time to first antibiotic modification was faster in the RAPID arm vs the SOC arm for overall antibiotics (8.6 [2.6–27.6] vs 14.9 [3.3–41.1] hours; P = .02) and gram-negative antibiotics (17.3 [4.9–72] vs 42.1 [10.1–72] hours; P < .001). Median (IQR) time to antibiotic escalation was faster in the RAPID arm vs the SOC arm for antimicrobial-resistant BSIs (18.4 [5.8–72] vs 61.7 [30.4–72] hours; P = .01). There were no differences between the arms in patient outcomes. Conclusions. Rapid organism ID and phenotypic AST led to faster changes in antibiotic therapy for gram-negative BSIs. 
    more » « less
  3. Abstract

    Rapid identification of antibiotic-resistant bacteria will play a key role in solving the global antibiotic crisis by providing a route to targeted antibiotic administration. However, current bacterial infection diagnoses take up to 3 days which can lead to antibiotic treatment that is less effective. Here we report a microfluidic system with a ladder shaped design allowing us to generate a twofold serial dilution of antibiotics comparable to current national and international standards. Our consolidated design, with minimal handling steps cuts down the testing time for antibiotic susceptibility from 16–20 h to 4–5 h. Our feasibility testing results are consistent with the commercial antibiotic susceptibility testing (AST) results, showing a 91.75% rate of agreement for Gram-negative and Gram-positive bacterial isolated from canine urinary tract infections (UTI) and may be used without prior isolation or enrichment. This platform provides an adaptable and efficient diagnostic tool for antibiotic susceptibility testing.

     
    more » « less
  4. Abstract

    Staphylococcus aureusis a Gram‐positive bacterium that colonizes almost every organ in humans and mice and is a leading cause of diseases worldwide.S. aureusinfections can be challenging to treat due to widespread antibiotic resistance and their ability to cause tissue damage. The primary modes of transmission ofS. aureusare via direct contact with a colonized or infected individual or invasive spread from a colonization niche in the same individual.S. aureuscan cause a myriad of diseases, including skin and soft tissue infections (SSTIs), osteomyelitis, pneumonia, endocarditis, and sepsis.S. aureusinfection is characterized by the formation of purulent lesions known as abscesses, which are rich in live and dead neutrophils, macrophages, and surrounded by a capsule containing fibrin and collagen. Different strains ofS. aureusproduce varying amounts of toxins that evade and/or elicit immune responses. Therefore, animal models ofS. aureusinfection provide a unique opportunity to understand the dynamics of organ‐specific immune responses and modifications in the pathogen that could favor the establishment of the pathogen. With advances in in vivo imaging of fluorescent transgenic mice, combined with fluorescent/bioluminescent bacteria, we can use mouse models to better understand the immune response to these types of infections. By understanding the host and bacterial dynamics within various organ systems, we can develop therapeutics to eliminate these pathogens. This module describes in vivo mouse models of both local and systemicS. aureusinfection. © 2021 Wiley Periodicals LLC.

    This article was corrected on 20 July 2022. See the end of the full text for details.

    Basic Protocol 1: Murine model ofStaphylococcus aureussubcutaneous infection

    Alternate Protocol: Murine tape stripping skin infection model

    Basic Protocol 2: Sample collection to determine skin structure, production of inflammatory mediators, and bacterial load

    Basic Protocol 3: Murine model of post‐traumaticStaphylococcus aureusosteomyelitis

    Basic Protocol 4: Intravenous infection of the retro‐orbital sinus

    Support Protocol: Preparation of the bacterial inoculum

     
    more » « less
  5. Abstract

    Each year, thousands of patients die from antimicrobial‐resistant bacterial infections that fail to respond to conventional antibiotic treatment. Antimicrobial polymers are a promising new method of combating antibiotic‐resistant bacterial infections. We have previously reported the synthesis of a series of narrow‐spectrum peptidomimetic antimicrobial polyurethanes that are effective against Gram‐negative bacteria, such asEscherichia coli; however, these polymers are not effective against Gram‐positive bacteria, such asStaphylococcus aureus. With the aim of understanding the correlation between chemical structure and antibacterial activity, we have subsequently developed three structural variants of these antimicrobial polyurethanes using post‐polymerization modification with decanoic acid and oleic acid. Our results show that such modifications converted the narrow‐spectrum antibacterial activity of these polymers into broad‐spectrum activity against Gram‐positive species such asS. aureus, however, also increasing their toxicity to mammalian cells. Mechanistic studies of bacterial membrane disruption illustrate the differences in antibacterial action between the various polymers. The results demonstrate the challenge of balancing antimicrobial activity and mammalian cell compatibility in the design of antimicrobial polymer compositions. © 2019 Society of Chemical Industry

     
    more » « less