2026-04-18T05:19:54Zhttps://recercat.cat/oai/request

oai:recercat.cat:10230/523172026-01-22T01:39:31Zcom_2072_6col_2072_452952

00925njm 22002777a 4500 dc Garrido, Victoria author Piñero-Lambea, Carlos author Rodriguez-Arce, Irene author Paetzold, Bernhard, 1981- author Ferrar, Tony author Weber, Marc author Garcia-Ramallo, Eva author Gallo López, Carolina, 1984- author Collantes, María author Peñuelas, Iván author Serrano Pubull, Luis, 1982- author Grilló, María-Jesús author Lluch-Senar, Maria 1982- author 2022-01-25T11:46:46Z 2022-01-25T11:46:46Z 2021 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. This work has been supported by the European Research Council (ERC) under the European Union’s Horizon 2020 research and innovation program, under grant agreement 670216 (MYCOCHASSIS). We also acknowledge the support of the Spanish Ministry of Economy, Industry and Competitiveness (MEIC) to the EMBL partnership, the Centro de Excelencia Severo Ochoa, the CERCA Program from the Generalitat de Catalunya, the European Union’s Horizon 2020 research and innovation program under grant agreement 634942 (MycoSynVac), and the La Caixa Fundation grant (Livetherapeutics HR18-00058). M.L.-S. acknowledges the support from FEDER project from Instituto Carlos III (ISCIII, Acción Estratégica en Salud 2016) (reference CP16/00094). We also acknowledge the staff of CRG/UPF Proteomics Unit, which is part of the Spanish Infrastructure for Omics Technologies (ICTS OmicsTech) unit is a member of the ProteoRed PRB3 consortium, which is supported by grant PT17/0019 of the PE I + D+i 2013-2016 from the Instituto de Salud Carlos III (ISCIII) and ERDF Bacteris -- Tractament Home -- Malalties Micoplasmosis Genòmica Engineering a genome-reduced bacterium to eliminate Staphylococcus aureus biofilms in vivo