ORIGINAL RESEARCH article
Front. Microbiol.
Sec. Antimicrobials, Resistance and Chemotherapy
Volume 16 - 2025 | doi: 10.3389/fmicb.2025.1660799
This article is part of the Research TopicAdvancing Antimicrobial Strategies: Nucleic Acid and Peptide-Based ApproachesView all 9 articles
Antisense Phosphorodiamidate Morpholino Oligomers Retain Activity in Burkholderia cepacia Complex Biofilm
Provisionally accepted
- 1University of Texas Southwestern Medical Center, Dallas, United States
- 2The University of Texas at Dallas, Richardson, United States
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Background. Members of the Burkholderia cepacia complex (Bcc) establish severe pulmonary infections in immunocompromised hosts, namely individuals with cystic fibrosis (CF) and chronic granulomatous disease (CGD). Due to innate antibiotic-resistant phenotypes and formation of protective biofilms, Bcc bacteria are difficult to eradicate from colonized lungs with traditional antibiotics. An alternative therapeutic approach involves the use of antisense molecules, peptideconjugated phosphorodiamidate morpholino oligomers (PPMOs). Previously, we found PPMOs targeting an acyl carrier protein (AcpP) reduce the burden of planktonic Bcc.Methods. Antimicrobial activities of AcpP PPMOs were assessed against established biofilms produced by Bcc clinical isolates, in which viable cells, biomass, and metabolic activity were enumerated. Bactericidal effects were further evaluated by microscopy. Cytotoxicity of these molecules was tested in human pulmonary cell lines.Results. AcpP PPMO treatment resulted in >3-log reductions (p <0.0001) in biofilm burden across five clinical isolates of Bcc tested and were dose-dependent (5-40 μΜ). This effect was visualized with confocal and scanning electron microscopy. We further demonstrated that PPMOs associate with bacterial cells in a time-dependent fashion using a fluorescently labeled AcpP PPMO with B. cenocepacia K56-2 DsRed. Finally, alveolar cells retained viability with AcpP PPMOs at bactericidal dosages.The Bcc biofilm setting is not a deterrent against PPMO delivery or antimicrobial activity. This is supported by the colocalization of AcpP PPMOs and cells, membrane destruction, loss of cell viability, and biomass reduction. Together, these data provide evidence that AcpP PPMOs are a promising therapeutic strategy in the treatment of Bcc infections.
Keywords: Antisense - oligonucleotide-drug conjugates, PPMOs, Burkholderia cepacia complex (Bcc), Biofilm, Antibiotic development
Received: 06 Jul 2025; Accepted: 29 Jul 2025.
Copyright: © 2025 Mendez, Pybus and Greenberg. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence: David Greenberg, University of Texas Southwestern Medical Center, Dallas, United States
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