title: Antimicrobial Nanoplexes meet Model Bacterial Membranes: the key role of Cardiolipin.
abstract: Antimicrobial resistance to traditional antibiotics is a crucial challenge of medical research. Oligonucleotide therapeutics, such as antisense or Transcription Factor Decoys (TFDs), have the potential to circumvent current resistance mechanisms by acting on novel targets. However, their full translation into clinical application requires efficient delivery strategies and fundamental comprehension of their interaction with target bacterial cells. To address these points, we employed a novel cationic bolaamphiphile that binds TFDs with high affinity to form self-assembled complexes (nanoplexes). Confocal microscopy revealed that nanoplexes efficiently transfect bacterial cells, consistently with biological efficacy on animal models. To understand the factors affecting the delivery process, liposomes with varying compositions, taken as model synthetic bilayers, were challenged with nanoplexes and investigated with Scattering and Fluorescence techniques. Thanks to the combination of results on bacteria and synthetic membrane models we demonstrate for the first time that the prokaryotic-enriched anionic lipid Cardiolipin (CL) plays a key-role in the TFDs delivery to bacteria. Moreover, we can hypothesize an overall TFD delivery mechanism, where bacterial membrane reorganization with permeability increase and release of the TFD from the nanoplexes are the main factors. These results will be of great benefit to boost the development of oligonucleotides-based antimicrobials of superior efficacy.
authors.affiliation: Procarta Biosystems Ltd, Norwich Innovation Centre, Norwich, UK.
authors.forename: Alejandro
authors.initials: A
authors.lastname: Marín-Menéndez
authors.affiliation: Department of Chemistry "Ugo Schiff" and CSGI, University of Florence, Firenze, Italy.
authors.forename: Costanza
authors.initials: C
authors.lastname: Montis
authors.affiliation: Procarta Biosystems Ltd, Norwich Innovation Centre, Norwich, UK.|Norwich Medical School, University of East Anglia, Norwich, UK.
authors.forename: Teresa
authors.initials: T
authors.lastname: Díaz-Calvo
authors.affiliation: Procarta Biosystems Ltd, Norwich Innovation Centre, Norwich, UK.
authors.forename: Davide
authors.initials: D
authors.lastname: Carta
authors.affiliation: Procarta Biosystems Ltd, Norwich Innovation Centre, Norwich, UK.|School of Pharmacy, University of East Anglia, Norwich, UK.
authors.forename: Kostas
authors.initials: K
authors.lastname: Hatzixanthis
authors.affiliation: School of Pharmacy, University of East Anglia, Norwich, UK.
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Morris
authors.affiliation: Procarta Biosystems Ltd, Norwich Innovation Centre, Norwich, UK.|Norwich Medical School, University of East Anglia, Norwich, UK.
authors.forename: Michael
authors.initials: M
authors.lastname: McArthur
authors.affiliation: Department of Chemistry "Ugo Schiff" and CSGI, University of Florence, Firenze, Italy.
authors.forename: Debora
authors.initials: D
authors.lastname: Berti
mesh_terms: D000818:Animals; D000890:Anti-Infective Agents / Q000494:pharmacology*; D018938:Caco-2 Cells; D002308:Cardiolipins / Q000378:metabolism*; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism*; D000067493:Dynamic Light Scattering; D004926:Escherichia coli / Q000187:drug effects / Q000378:metabolism*; D005452:Fluoresceins / Q000378:metabolism; D005663:Furans / Q000737:chemistry; D006801:Humans; D008081:Liposomes; D008647:Mesocricetus; D008954:Models, Biological; D053758:Nanoparticles / Q000737:chemistry*; D011728:Pyridones / Q000737:chemistry; D017207:Rats, Sprague-Dawley
chemical_list: D000890:Anti-Infective Agents; D002308:Cardiolipins; D005452:Fluoresceins; D005663:Furans; D008081:Liposomes; D011728:Pyridones; C543575:bolaamphiphile; C024098:6-carboxyfluorescein