Engineering a genome‐reduced bacterium to eliminate Staphylococcus aureus biofilms in vivo

Abstract Bacteria present a promising delivery system for treating human diseases. Here, we engineered the genome‐reduced human lung pathogen Mycoplasma pneumoniae as a live biotherapeutic to treat biofilm‐associated bacterial infections. This strain has a unique genetic code, which hinders gene tra...

Full description

Saved in:
Bibliographic Details
Main Authors: Victoria Garrido, Carlos Piñero‐Lambea, Irene Rodriguez‐Arce, Bernhard Paetzold, Tony Ferrar, Marc Weber, Eva Garcia‐Ramallo, Carolina Gallo, María Collantes, Iván Peñuelas, Luis Serrano, María‐Jesús Grilló, María Lluch‐Senar
Format: Article
Language:English
Published: Springer Nature 2021-10-01
Series:Molecular Systems Biology
Subjects:
Online Access:https://doi.org/10.15252/msb.202010145
Tags: Add Tag
No Tags, Be the first to tag this record!
_version_ 1849389089132904448
author Victoria Garrido
Carlos Piñero‐Lambea
Irene Rodriguez‐Arce
Bernhard Paetzold
Tony Ferrar
Marc Weber
Eva Garcia‐Ramallo
Carolina Gallo
María Collantes
Iván Peñuelas
Luis Serrano
María‐Jesús Grilló
María Lluch‐Senar
author_facet Victoria Garrido
Carlos Piñero‐Lambea
Irene Rodriguez‐Arce
Bernhard Paetzold
Tony Ferrar
Marc Weber
Eva Garcia‐Ramallo
Carolina Gallo
María Collantes
Iván Peñuelas
Luis Serrano
María‐Jesús Grilló
María Lluch‐Senar
author_sort Victoria Garrido
collection DOAJ
description Abstract Bacteria present a promising delivery system for treating human diseases. Here, we engineered the genome‐reduced human lung pathogen Mycoplasma pneumoniae as a live biotherapeutic to treat biofilm‐associated bacterial infections. This strain has a unique genetic code, which hinders gene transfer to most other bacterial genera, and it lacks a cell wall, which allows it to express proteins that target peptidoglycans of pathogenic bacteria. We first determined that removal of the pathogenic factors fully attenuated the chassis strain in vivo. We then designed synthetic promoters and identified an endogenous peptide signal sequence that, when fused to heterologous proteins, promotes efficient secretion. Based on this, we equipped the chassis strain with a genetic platform designed to secrete antibiofilm and bactericidal enzymes, resulting in a strain capable of dissolving Staphylococcus aureus biofilms preformed on catheters in vitro, ex vivo, and in vivo. To our knowledge, this is the first engineered genome‐reduced bacterium that can fight against clinically relevant biofilm‐associated bacterial infections.
format Article
id doaj-art-e81cf3839c824b17817201e7486d0d94
institution Kabale University
issn 1744-4292
language English
publishDate 2021-10-01
publisher Springer Nature
record_format Article
series Molecular Systems Biology
spelling doaj-art-e81cf3839c824b17817201e7486d0d942025-08-20T03:42:04ZengSpringer NatureMolecular Systems Biology1744-42922021-10-01171012010.15252/msb.202010145Engineering a genome‐reduced bacterium to eliminate Staphylococcus aureus biofilms in vivoVictoria Garrido0Carlos Piñero‐Lambea1Irene Rodriguez‐Arce2Bernhard Paetzold3Tony Ferrar4Marc Weber5Eva Garcia‐Ramallo6Carolina Gallo7María Collantes8Iván Peñuelas9Luis Serrano10María‐Jesús Grilló11María Lluch‐Senar12Centre for Genomic Regulation (CRG), The Barcelona Institute of Science and TechnologyCentre for Genomic Regulation (CRG), The Barcelona Institute of Science and TechnologyCentre for Genomic Regulation (CRG), The Barcelona Institute of Science and TechnologyCentre for Genomic Regulation (CRG), The Barcelona Institute of Science and TechnologyCentre for Genomic Regulation (CRG), The Barcelona Institute of Science and TechnologyCentre for Genomic Regulation (CRG), The Barcelona Institute of Science and TechnologyCentre for Genomic Regulation (CRG), The Barcelona Institute of Science and TechnologyCentre for Genomic Regulation (CRG), The Barcelona Institute of Science and TechnologyRADIOMIN Research Group, Clínica Universidad de NavarraRADIOMIN Research Group, Clínica Universidad de NavarraCentre for Genomic Regulation (CRG), The Barcelona Institute of Science and TechnologyInstitute of Agrobiotechnology (IdAB; CSIC‐Navarra Government)Centre for Genomic Regulation (CRG), The Barcelona Institute of Science and TechnologyAbstract Bacteria present a promising delivery system for treating human diseases. Here, we engineered the genome‐reduced human lung pathogen Mycoplasma pneumoniae as a live biotherapeutic to treat biofilm‐associated bacterial infections. This strain has a unique genetic code, which hinders gene transfer to most other bacterial genera, and it lacks a cell wall, which allows it to express proteins that target peptidoglycans of pathogenic bacteria. We first determined that removal of the pathogenic factors fully attenuated the chassis strain in vivo. We then designed synthetic promoters and identified an endogenous peptide signal sequence that, when fused to heterologous proteins, promotes efficient secretion. Based on this, we equipped the chassis strain with a genetic platform designed to secrete antibiofilm and bactericidal enzymes, resulting in a strain capable of dissolving Staphylococcus aureus biofilms preformed on catheters in vitro, ex vivo, and in vivo. To our knowledge, this is the first engineered genome‐reduced bacterium that can fight against clinically relevant biofilm‐associated bacterial infections.https://doi.org/10.15252/msb.202010145bacterial therapybiofilmin vivoMycoplasmasynthetic biology
spellingShingle Victoria Garrido
Carlos Piñero‐Lambea
Irene Rodriguez‐Arce
Bernhard Paetzold
Tony Ferrar
Marc Weber
Eva Garcia‐Ramallo
Carolina Gallo
María Collantes
Iván Peñuelas
Luis Serrano
María‐Jesús Grilló
María Lluch‐Senar
Engineering a genome‐reduced bacterium to eliminate Staphylococcus aureus biofilms in vivo
Molecular Systems Biology
bacterial therapy
biofilm
in vivo
Mycoplasma
synthetic biology
title Engineering a genome‐reduced bacterium to eliminate Staphylococcus aureus biofilms in vivo
title_full Engineering a genome‐reduced bacterium to eliminate Staphylococcus aureus biofilms in vivo
title_fullStr Engineering a genome‐reduced bacterium to eliminate Staphylococcus aureus biofilms in vivo
title_full_unstemmed Engineering a genome‐reduced bacterium to eliminate Staphylococcus aureus biofilms in vivo
title_short Engineering a genome‐reduced bacterium to eliminate Staphylococcus aureus biofilms in vivo
title_sort engineering a genome reduced bacterium to eliminate staphylococcus aureus biofilms in vivo
topic bacterial therapy
biofilm
in vivo
Mycoplasma
synthetic biology
url https://doi.org/10.15252/msb.202010145
work_keys_str_mv AT victoriagarrido engineeringagenomereducedbacteriumtoeliminatestaphylococcusaureusbiofilmsinvivo
AT carlospinerolambea engineeringagenomereducedbacteriumtoeliminatestaphylococcusaureusbiofilmsinvivo
AT irenerodriguezarce engineeringagenomereducedbacteriumtoeliminatestaphylococcusaureusbiofilmsinvivo
AT bernhardpaetzold engineeringagenomereducedbacteriumtoeliminatestaphylococcusaureusbiofilmsinvivo
AT tonyferrar engineeringagenomereducedbacteriumtoeliminatestaphylococcusaureusbiofilmsinvivo
AT marcweber engineeringagenomereducedbacteriumtoeliminatestaphylococcusaureusbiofilmsinvivo
AT evagarciaramallo engineeringagenomereducedbacteriumtoeliminatestaphylococcusaureusbiofilmsinvivo
AT carolinagallo engineeringagenomereducedbacteriumtoeliminatestaphylococcusaureusbiofilmsinvivo
AT mariacollantes engineeringagenomereducedbacteriumtoeliminatestaphylococcusaureusbiofilmsinvivo
AT ivanpenuelas engineeringagenomereducedbacteriumtoeliminatestaphylococcusaureusbiofilmsinvivo
AT luisserrano engineeringagenomereducedbacteriumtoeliminatestaphylococcusaureusbiofilmsinvivo
AT mariajesusgrillo engineeringagenomereducedbacteriumtoeliminatestaphylococcusaureusbiofilmsinvivo
AT marialluchsenar engineeringagenomereducedbacteriumtoeliminatestaphylococcusaureusbiofilmsinvivo