Nathalie Weibel, Martina Curcio, Atilla Schreiber, Gabriel Arriaga, Marine Mausy, Jana Mehdy, Lea Brüllmann, Andreas Meyer, Len Roth, Tamara Flury, Valerie Pecina, Kim Starlinger, Jan Dernič, Kenny Jungfer, Fabian Ackle, Jennifer Earp, Martin Hausmann, Martin Jinek, Gerhard Rogler, Cauã Antunes Westmann
{"title":"在大肠杆菌 Nissle 1917 中设计新型益生菌工具包,用于感知和缓解肠道炎症性疾病。","authors":"Nathalie Weibel, Martina Curcio, Atilla Schreiber, Gabriel Arriaga, Marine Mausy, Jana Mehdy, Lea Brüllmann, Andreas Meyer, Len Roth, Tamara Flury, Valerie Pecina, Kim Starlinger, Jan Dernič, Kenny Jungfer, Fabian Ackle, Jennifer Earp, Martin Hausmann, Martin Jinek, Gerhard Rogler, Cauã Antunes Westmann","doi":"10.1021/acssynbio.4c00036","DOIUrl":null,"url":null,"abstract":"<p><p>Inflammatory bowel disease (IBD) is characterized by chronic intestinal inflammation with no cure and limited treatment options that often have systemic side effects. In this study, we developed a target-specific system to potentially treat IBD by engineering the probiotic bacterium <i>Escherichia coli</i> <i>Nissle 1917</i> (EcN). Our modular system comprises three components: a transcription factor-based sensor (NorR) capable of detecting the inflammation biomarker nitric oxide (NO), a type 1 hemolysin secretion system, and a therapeutic cargo consisting of a library of humanized anti-TNFα nanobodies. Despite a reduction in sensitivity, our system demonstrated a concentration-dependent response to NO, successfully secreting functional nanobodies with binding affinities comparable to the commonly used drug Adalimumab, as confirmed by enzyme-linked immunosorbent assay and in vitro assays. This newly validated nanobody library expands EcN therapeutic capabilities. The adopted secretion system, also characterized for the first time in EcN, can be further adapted as a platform for screening and purifying proteins of interest. Additionally, we provided a mathematical framework to assess critical parameters in engineering probiotic systems, including the production and diffusion of relevant molecules, bacterial colonization rates, and particle interactions. This integrated approach expands the synthetic biology toolbox for EcN-based therapies, providing novel parts, circuits, and a model for tunable responses at inflammatory hotspots.</p>","PeriodicalId":26,"journal":{"name":"ACS Synthetic Biology","volume":null,"pages":null},"PeriodicalIF":3.7000,"publicationDate":"2024-08-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11334186/pdf/","citationCount":"0","resultStr":"{\"title\":\"Engineering a Novel Probiotic Toolkit in <i>Escherichia coli</i> <i>Nissle 1917</i> for Sensing and Mitigating Gut Inflammatory Diseases.\",\"authors\":\"Nathalie Weibel, Martina Curcio, Atilla Schreiber, Gabriel Arriaga, Marine Mausy, Jana Mehdy, Lea Brüllmann, Andreas Meyer, Len Roth, Tamara Flury, Valerie Pecina, Kim Starlinger, Jan Dernič, Kenny Jungfer, Fabian Ackle, Jennifer Earp, Martin Hausmann, Martin Jinek, Gerhard Rogler, Cauã Antunes Westmann\",\"doi\":\"10.1021/acssynbio.4c00036\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>Inflammatory bowel disease (IBD) is characterized by chronic intestinal inflammation with no cure and limited treatment options that often have systemic side effects. 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Engineering a Novel Probiotic Toolkit in Escherichia coliNissle 1917 for Sensing and Mitigating Gut Inflammatory Diseases.
Inflammatory bowel disease (IBD) is characterized by chronic intestinal inflammation with no cure and limited treatment options that often have systemic side effects. In this study, we developed a target-specific system to potentially treat IBD by engineering the probiotic bacterium Escherichia coliNissle 1917 (EcN). Our modular system comprises three components: a transcription factor-based sensor (NorR) capable of detecting the inflammation biomarker nitric oxide (NO), a type 1 hemolysin secretion system, and a therapeutic cargo consisting of a library of humanized anti-TNFα nanobodies. Despite a reduction in sensitivity, our system demonstrated a concentration-dependent response to NO, successfully secreting functional nanobodies with binding affinities comparable to the commonly used drug Adalimumab, as confirmed by enzyme-linked immunosorbent assay and in vitro assays. This newly validated nanobody library expands EcN therapeutic capabilities. The adopted secretion system, also characterized for the first time in EcN, can be further adapted as a platform for screening and purifying proteins of interest. Additionally, we provided a mathematical framework to assess critical parameters in engineering probiotic systems, including the production and diffusion of relevant molecules, bacterial colonization rates, and particle interactions. This integrated approach expands the synthetic biology toolbox for EcN-based therapies, providing novel parts, circuits, and a model for tunable responses at inflammatory hotspots.
期刊介绍:
The journal is particularly interested in studies on the design and synthesis of new genetic circuits and gene products; computational methods in the design of systems; and integrative applied approaches to understanding disease and metabolism.
Topics may include, but are not limited to:
Design and optimization of genetic systems
Genetic circuit design and their principles for their organization into programs
Computational methods to aid the design of genetic systems
Experimental methods to quantify genetic parts, circuits, and metabolic fluxes
Genetic parts libraries: their creation, analysis, and ontological representation
Protein engineering including computational design
Metabolic engineering and cellular manufacturing, including biomass conversion
Natural product access, engineering, and production
Creative and innovative applications of cellular programming
Medical applications, tissue engineering, and the programming of therapeutic cells
Minimal cell design and construction
Genomics and genome replacement strategies
Viral engineering
Automated and robotic assembly platforms for synthetic biology
DNA synthesis methodologies
Metagenomics and synthetic metagenomic analysis
Bioinformatics applied to gene discovery, chemoinformatics, and pathway construction
Gene optimization
Methods for genome-scale measurements of transcription and metabolomics
Systems biology and methods to integrate multiple data sources
in vitro and cell-free synthetic biology and molecular programming
Nucleic acid engineering.