Engineering a Novel Probiotic Toolkit in Escherichia coli Nissle 1917 for Sensing and Mitigating Gut Inflammatory Diseases

被引：2

作者：

Weibel, Nathalie ^{[1
]}

Curcio, Martina ^{[1
]}

Schreiber, Atilla ^{[1
]}

Arriaga, Gabriel ^{[1
]}

Mausy, Marine ^{[1
]}

Mehdy, Jana ^{[1
]}

Brullmann, Lea ^{[1
]}

Meyer, Andreas ^{[1
]}

Roth, Len ^{[1
]}

Flury, Tamara ^{[1
]}

Pecina, Valerie ^{[1
]}

Starlinger, Kim ^{[1
]}

Dernic, Jan ^{[2
]}

Jungfer, Kenny ^{[3
]}

Ackle, Fabian ^{[4
]}

Earp, Jennifer ^{[4
]}

Hausmann, Martin ^{[5
,6
]}

Jinek, Martin ^{[3
]}

Rogler, Gerhard ^{[5
,6
]}

Westmann, Caua Antunes ^{[7
,8
]}

机构：

[1] Univ Zurich, CH-8057 Zurich, Switzerland

[2] Univ Zurich, Inst Pharmacol & Toxicol, CH-8057 Zurich, Switzerland

[3] Univ Zurich, Dept Biochem, CH-8057 Zurich, Switzerland

[4] Univ Zurich, Inst Med Microbiol, CH-8006 Zurich, Switzerland

[5] Univ Hosp Zurich, Dept Gastroenterol & Hepatol, CH-8091 Zurich, Switzerland

[6] Zurich Univ, CH-8091 Zurich, Switzerland

[7] Univ Zurich, Dept Evolutionary Biol & Environm Studies, CH-8057 Zurich, Switzerland

[8] Swiss Inst Bioinformat, CH-1015 Lausanne, Switzerland

来源：

ACS SYNTHETIC BIOLOGY | 2024年 / 13卷 / 08期

关键词：

engineered probiotic; IBD; inflammation; E. coli Nissle 1917 (EcN); nitric oxide; TNF alpha; nanobodies; NITRIC-OXIDE; SYNTHETIC BIOLOGY; ULCERATIVE-COLITIS; BOWEL-DISEASE; IN-SITU; PROTEIN; SECRETION; NORR; TNF; SIMULATION;

D O I：

10.1021/acssynbio.4c00036

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

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 coli Nissle 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-TNF alpha 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.

引用

页码：2376 / 2390

页数：15

共 105 条

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