Tissue nano-transfection of antimicrobial genes drives bacterial biofilm killing in wounds and is potentially mediated by extracellular vesicles

Tatiana Z Cuellar-Gaviria; Maria Angelica Rincon-Benavides; Hatice Nur Halipci Topsakal; Ana Isabel Salazar-Puerta; Shara Jaramillo-Garrido; Mia Kordowski; Carlos A Vasquez-Martinez; Kim Truc Nguyen; Xilal Y Rima; Pranav S J B Rana; Orlando Combita-Heredia; Binbin Deng; Kavya Dathathreya; David W McComb; Eduardo Reategui; Daniel Wozniak; Natalia Higuita-Castro; Daniel Gallego-Perez

doi:10.1016/j.jconrel.2024.10.071

Tissue nano-transfection of antimicrobial genes drives bacterial biofilm killing in wounds and is potentially mediated by extracellular vesicles

J Control Release. 2024 Nov 13:376:1300-1315. doi: 10.1016/j.jconrel.2024.10.071. Online ahead of print.

Authors

Tatiana Z Cuellar-Gaviria¹, Maria Angelica Rincon-Benavides², Hatice Nur Halipci Topsakal³, Ana Isabel Salazar-Puerta⁴, Shara Jaramillo-Garrido⁴, Mia Kordowski², Carlos A Vasquez-Martinez⁵, Kim Truc Nguyen⁶, Xilal Y Rima⁶, Pranav S J B Rana⁷, Orlando Combita-Heredia⁴, Binbin Deng⁸, Kavya Dathathreya⁴, David W McComb⁹, Eduardo Reategui¹⁰, Daniel Wozniak¹¹, Natalia Higuita-Castro¹², Daniel Gallego-Perez¹³

Affiliations

¹ Department of Biomedical Engineering, The Ohio State University, Columbus, OH 43210, USA; Dorothy M. Davis Heart and Lung Research Institute, The Ohio State University, Columbus, OH 43210, USA; Gene Therapy Institute, The Ohio State University, Columbus, OH 43210, USA; Infectious Disease Institute, The Ohio State University, Columbus, OH 43210, USA.
² Department of Biomedical Engineering, The Ohio State University, Columbus, OH 43210, USA; Biophysics Program, The Ohio State University, Columbus, OH 43210, USA.
³ Department of Biomedical Engineering, The Ohio State University, Columbus, OH 43210, USA; Istanbul Atlas University, Istanbul 34408, Turkiye.
⁴ Department of Biomedical Engineering, The Ohio State University, Columbus, OH 43210, USA.
⁵ Department of Biomedical Engineering, The Ohio State University, Columbus, OH 43210, USA; CONACYT - Faculty of Medicine, Benito Juárez Autonomous University of Oaxaca, Oaxaca 68020, Mexico.
⁶ William G. Lowrie Department of Chemical and Biomolecular Engineering, The Ohio State University, Columbus, OH 43210, USA.
⁷ Department of Microbiology, The Ohio State University, Columbus, OH 43210, USA; Department of Microbial Infection and Immunity, The Ohio State University, Columbus, OH 43210, USA.
⁸ Center for Electron Microscopy and Analysis (CEMAS), The Ohio State University, Columbus, OH 43210, USA.
⁹ Center for Electron Microscopy and Analysis (CEMAS), The Ohio State University, Columbus, OH 43210, USA; Department of Materials Science and Engineering, The Ohio State University, Columbus, OH 43210, USA.
¹⁰ William G. Lowrie Department of Chemical and Biomolecular Engineering, The Ohio State University, Columbus, OH 43210, USA; Comprehensive Cancer Center, The Ohio State University, Columbus, OH 43210, USA.
¹¹ Infectious Disease Institute, The Ohio State University, Columbus, OH 43210, USA; Department of Microbiology, The Ohio State University, Columbus, OH 43210, USA; Department of Microbial Infection and Immunity, The Ohio State University, Columbus, OH 43210, USA.
¹² Department of Biomedical Engineering, The Ohio State University, Columbus, OH 43210, USA; Dorothy M. Davis Heart and Lung Research Institute, The Ohio State University, Columbus, OH 43210, USA; Gene Therapy Institute, The Ohio State University, Columbus, OH 43210, USA; Infectious Disease Institute, The Ohio State University, Columbus, OH 43210, USA; Biophysics Program, The Ohio State University, Columbus, OH 43210, USA; Department of Neurological Surgery, The Ohio State University, Columbus, OH 43210, USA.
¹³ Department of Biomedical Engineering, The Ohio State University, Columbus, OH 43210, USA; Dorothy M. Davis Heart and Lung Research Institute, The Ohio State University, Columbus, OH 43210, USA; Gene Therapy Institute, The Ohio State University, Columbus, OH 43210, USA; Infectious Disease Institute, The Ohio State University, Columbus, OH 43210, USA; Biophysics Program, The Ohio State University, Columbus, OH 43210, USA; Department of Surgery, The Ohio State University, Columbus, OH 43210, USA. Electronic address: [email protected].

PMID: 39491627
DOI: 10.1016/j.jconrel.2024.10.071

Abstract

The emergence of bacteria that are resistant to antibiotics is on track to become a major global health crisis. Therefore, there is an urgent need for new treatment options. Here, we studied the implementation of tissue-nanotransfection (TNT) to treat Staphylococcus aureus-infected wounds by delivering gene cargos that boost the levels of naturally produced antimicrobial peptides. The Cathelicidin Antimicrobial Peptide gene (CAMP), which produces the antimicrobial peptide LL-37, was used as model gene cargo. In vitro evaluation showed successful transfection and an increase in the transcription and translation of CAMP-coding plasmid in mouse primary epithelial cells. Moreover, we found that the extracellular vesicles (EVs) derived from the transfected cells (in vitro and in vivo) carried significantly higher concentrations of CAMP transcripts and LL-37 peptide compared to control EVs, possibly mediating the trafficking of the antimicrobial contents to other neighboring cells. The TNT platform was then used in vivo on an excisional wound model in mice to nanotransfect the CAMP-coding plasmid on the edge of infected wounds. After 4 days of daily treatment, we observed a significant decrease in the bacterial load in the CAMP-treated group compared to the sham group. Moreover, histological analysis and bacterial load quantification also revealed that TNT of CAMP on S. aureus-infected wounds was effective in treating biofilm progression by reducing the bacterial load. Lastly, we observed a significant increase in macrophage recruitment to the infected tissue, a robust increase in vascularization, as well as and an increased expression of IL10 and Fli1. Our results demonstrate that TNT-based delivery of gene cargos coding for antimicrobial compounds to the wound is a promising approach for combating biofilm infections in wounds.

Keywords: Bacterial infections; Biofilms; CAMP/CRAMP; Extracellular vesicles (EVs); LL-37/CRAMP; Staphylococcus aureus; Tissue nano-transfection (TNT).