Enginyeria d'espècies de Mycoplasma per a aplicacions biotecnològiques i biomèdiques

Ariadna Montero Blay

Prof. Luis Serrano Pubul Dr. Carlos Piñero Lambea

Prof. Luis Serrano Pubul i Dr. Carlos Piñero Lambea

Mycoplasmas are a group of bacteria characterized by minimized genomes, limited biosynthetic capacities, and simplified metabolic networks. In this work, we explored how Mycoplasma species can be exploited for biotechnological and therapeutic applications. In chapter two, we engineered M. pneumoniae transposons so that they could efficiently transform different Mycoplasma species. This fact allows the generation of essentiality studies in these species, a critical analysis to identify genes that could be deleted to create attenuated vaccination strains. Chapter three developed a new method that uses high-resolution transposon and proteomics data to infer active metabolic pathways within a cell at a specific moment. This information can be used when engineering attenuated strains. In chapter four, we explored the capacity of M. pneumoniae to express functional human biologics in vitro and in mice lungs. Chapter five used the protein design algorithm FoldX and ModelX to mutate a human interleukin to enhance its properties in terms of affinity to its receptors and increased bacterial expression. Chapter six identified the secretion signals and designed synthetic promoters in the recent-discovered fast-growing Mycoplasma feriruminatoris. Altogether this thesis develops the tools for exploiting Mycoplasmas for biotechnological purposes (M. agalactiae, M. feriruminatoris) and validates M. pneumoniae for human lung therapy immunomodulation.

En resum, aquesta tesi proposa primer eines genètiques per modificar bacteris per generar vacunes per al bestiar. A la segona part d'aquesta tesi, s'explota el bacteri M. pneumoniae per tractar malalties pulmonars. Utilitzant aquest vehicle bacterià i modificant una interleucina antiinflamatòria per optimitzar les seves propietats, ha estat possible eliminar completament la inflamació pulmonar causada per la infecció amb un altre bacteri (treball sota patent i en revisió a PNAS). El treball d'aquesta tesi, que tracta de l'explotació de M. pneumoniae com a sistema d'expressió biològica in vivo, podria ser llicenciat a l'empresa spin-off Pulmobiotics o altres en el camp de les teràpies mediades per virus o bacteris. A més, el treball d'enginyeria de molècules per se (modificació de citocines per optimitzar les seves propietats amb finalitats terapèutiques) es pot aplicar a altres citocines amb similituds estructurals. Es pretén que aquest treball s'exploti a través d'una nova spin-off, de la qual Ariadna Montero serà una de les cofundadores, sent també coinventora en les diferents patents en curs o previstes.

Mycoplasmas; Minimized Genomes; Biosynthetic Capacities; Simplified Metabolic Networks; Biotechnological Applications; Therapeutic Applications; M. pneumoniae Transposons; Efficient Transformation; Mycoplasma Species; Essentiality Studies; Attenuated Vaccination Strains; High-Resolution Transposon Data; Proteomics Data; Active Metabolic Pathways; Attenuated Strains; Functional Human Biologics; In Vitro Expression; Mice Lungs; Protein Design Algorithm; FoldX; ModelX; Human Interleukin; Affinity Enhancement; Receptor Affinity; Increased Bacterial Expression; Secretion Signals; Synthetic Promoters; Fast-Growing Mycoplasma feriruminatoris; M. agalactiae; Human Lung Therapy Immunomodulation.