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

Szilamer Ferenczi; Norbert Solymosi; István Horváth; Natália Szeőcs; Zsuzsanna Grózer; Dániel Kuti; Balázs Juhász; Zsuzsanna Winkler; Tibor Pankotai; Farkas Sükösd; Anikó Stágel; Melinda Paholcsek; Dávid Dóra; Nándor Nagy; Krisztina J Kovács; Ivan Zanoni; Zoltan Szallasi

doi:10.1016/j.omtm.2020.11.010

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

Mol Ther Methods Clin Dev. 2020 Nov 20:20:218-226. doi: 10.1016/j.omtm.2020.11.010. eCollection 2021 Mar 12.

Authors

Szilamer Ferenczi^{1

2}, Norbert Solymosi³, István Horváth², Natália Szeőcs², Zsuzsanna Grózer², Dániel Kuti¹, Balázs Juhász¹, Zsuzsanna Winkler¹, Tibor Pankotai⁴, Farkas Sükösd⁵, Anikó Stágel⁶, Melinda Paholcsek⁶, Dávid Dóra⁷, Nándor Nagy⁷, Krisztina J Kovács¹, Ivan Zanoni⁸, Zoltan Szallasi^{9

10

11}

Affiliations

¹ Institute of Experimental Medicine, Laboratory of Molecular Neuroendocrinology, Budapest, Hungary.
² Central European Biosystems, Gödöllő, Hungary.
³ Centre for Bioinformatics, University of Veterinary Medicine Budapest, Budapest, Hungary.
⁴ Department of Biochemistry and Molecular Biology, Faculty of Science and Informatics, University of Szeged, Szeged, Hungary.
⁵ Department of Pathology, Laboratory of Molecular Pathology, Faculty of Medicine, University of Szeged, Szeged, Hungary.
⁶ Department of Human Genetics, Faculty of Medicine, University of Debrecen, Debrecen, Hungary.
⁷ Department of Anatomy, Faculty of Medicine, Histology and Embryology, Semmelweis University, Budapest, Hungary.
⁸ Divisions of Immunology and Gastroenterology, Harvard Medical School, Boston Children's Hospital, Boston, MA, USA.
⁹ Danish Cancer Society Research Center, Copenhagen, Denmark.
¹⁰ Computational Health Informatics Program, Boston Children's Hospital, Boston, MA, USA.
¹¹ 2nd Department of Pathology, MTA-SE NAP, Brain Metastasis Research Group, Hungarian Academy of Sciences, Semmelweis University, Budapest, Hungary.

Abstract

We developed an orally administered, engineered, bacterium-based, 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.

Keywords: RNA interference; bacterial therapeutic delivery; engineered E. coli, treatment of ulcerative colitis.

Grants and funding

P30 DK063608/DK/NIDDK NIH HHS/United States