Efficient treatment of a preclinical inflammatory bowel disease model with engineered bacteria

Abstract

We developed an orally administered, engineered, bacteriumbased, RNA interference-mediated therapeutic method to significantly reduce the symptoms in the most frequently used animal model of inflammatory bowel disease. This bacterium-mediated RNA interference strategy was based on the genomically stable, non-pathogenic E. coli MDS42 strain, which was engineered to constitutively produce invasin and the listeriolysin O cytolysin. These proteins enabled the bacteria first to invade the colon epithelium and then degrade in the phagosome. This allowed the delivery of a plasmid encoding small hairpin RNA (shRNA) targeting tumor necrosis factor (TNF) into the cytoplasm of the target cells. The expression levels of TNF and other cytokines significantly decreased upon this treatment in dextran sulfate sodium (DSS)-induced colitis, and the degree of inflammation was significantly reduced. With further safety modifications this method could serve as a safe and side effect-free alternative to biologicals targeting TNF or other inflammatory mediators.

Evidence of a Myenteric Plexus Barrier and Its Macrophage-Dependent Degradation During Murine Colitis: Implications in Enteric Neuroinflammation

“…Double immunofluorescence was performed to reveal the cell populations that express TNF, a master regulator of the inflammatory response. After induction of DSS colitis, TNF protein is expressed by the mucosal epithelial lining 37 ( Figure 10 K , dashed line), serosal F4/80+ cells ( Figure 10 K , arrowheads; and Figure 10 L , asterisks), clusters of mucosal macrophages ( Figure 10 M ), and enteric ganglia ( Figure 10 K and N , dashed line). Interestingly, F4/80+ MyMs ( Figure 10 K and N , arrows) and IGMs ( Figure 10 N , asterisk) show no co-localization with TNF expression.…”

Bioinspired and biomimetic strategies for inflammatory bowel disease therapy

“…183 Ferenczi et al developed an oral, engineered, bacterial-based, RNA interference-mediated therapeutic approach that demonstrated effective treatment for preclinical inflammatory bowel disease models with engineered bacteria. 184 Genetically engineered bacteria endowed with the function of creating a fibrous matrix capable of promoting intestinal epithelial integrity in situ by EcN were created by Praveschotinunt et al to facilitate the in situ production of therapeutic protein matrices utilizing beneficial bacteria. 185…”

Engineered Bacteria-Based Living Materials for Biotherapeutic Applications

“…While the inflammation was also decreased in DSS-induced colitis. Reproduced with permission from Ferenczi et al (2021) , Copyright 2021, Elsevier. (C) (i) The curli fiber variants produced from engineered E. coli Nissle at 37°C in LB media containing arabinose (Ara) measured by Congo Red Binding Assay.…”

“…Such steps cause a significant reduction of TNF and other cytokines and meanwhile also reduce the inflammation in DSS-induced colitis. This strategy is safe with no side effects so it could be used as a modern approach to target TNF and other inflammatory mediators ( Ferenczi et al, 2021 ). Recently, An et al (2020) designed a living glue system by inspiring the quality of self-regeneration of organisms.…”