Engineered Escherichia coli for the in situ secretion of therapeutic nanobodies in the gut

Jason P Lynch; Coral González-Prieto; Analise Z Reeves; Sena Bae; Urmila Powale; Neha P Godbole; Jacqueline M Tremblay; Florian I Schmidt; Hidde L Ploegh; Vikram Kansra; Jonathan N Glickman; John M Leong; Charles B Shoemaker; Wendy S Garrett; Cammie F Lesser

doi:10.1016/j.chom.2023.03.007

Engineered Escherichia coli for the in situ secretion of therapeutic nanobodies in the gut

Cell Host Microbe. 2023 Apr 12;31(4):634-649.e8. doi: 10.1016/j.chom.2023.03.007. Epub 2023 Mar 31.

Authors

Affiliations

¹ Center for Bacterial Pathogenesis, Division of Infectious Diseases, Department of Medicine, Massachusetts General Hospital, Boston, MA 02115, USA; Department of Microbiology, Blavatnik Institute, Harvard Medical School, Boston, MA 02115, USA.
² Departments of Immunology and Infectious Diseases and Harvard T.H. Chan Center for the Microbiome in Public Health, Harvard T.H. Chan School of Public Health, Boston, MA 02115, USA.
³ Department of Infectious Disease and Global Health, Tufts Cummings School of Veterinary Medicine, North Grafton, MA 01536, USA.
⁴ Institute of Innate Immunity, Medical Faculty, University of Bonn, 53127 Bonn, Germany.
⁵ Boston Children's Hospital and Harvard Medical School, Boston, MA 02115, USA.
⁶ Vicero Inc., Shrewsbury, MA 01545, USA.
⁷ Department of Pathology, Massachusetts General Hospital, Boston, MA 02115, USA.
⁸ Department of Molecular Biology and Microbiology, Tufts University School of Medicine, Boston, MA 02111, USA; Tufts Stuart B Levy Center for Integrated Management of Antimicrobial Resistance, Tufts University, Boston, MA 02111, USA.
⁹ Department of Infectious Disease and Global Health, Tufts Cummings School of Veterinary Medicine, North Grafton, MA 01536, USA; Department of Molecular Biology and Microbiology, Tufts University School of Medicine, Boston, MA 02111, USA.
¹⁰ Departments of Immunology and Infectious Diseases and Harvard T.H. Chan Center for the Microbiome in Public Health, Harvard T.H. Chan School of Public Health, Boston, MA 02115, USA; Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA 02215, USA; Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA.
¹¹ Center for Bacterial Pathogenesis, Division of Infectious Diseases, Department of Medicine, Massachusetts General Hospital, Boston, MA 02115, USA; Department of Microbiology, Blavatnik Institute, Harvard Medical School, Boston, MA 02115, USA; Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA; Ragon Institute of Harvard and MIT, Cambridge, MA 02139, USA. Electronic address: clesser@mgh.harvard.edu.

Abstract

Drug platforms that enable the directed delivery of therapeutics to sites of diseases to maximize efficacy and limit off-target effects are needed. Here, we report the development of PROT₃EcT, a suite of commensal Escherichia coli engineered to secrete proteins directly into their surroundings. These bacteria consist of three modular components: a modified bacterial protein secretion system, the associated regulatable transcriptional activator, and a secreted therapeutic payload. PROT₃EcT secrete functional single-domain antibodies, nanobodies (Nbs), and stably colonize and maintain an active secretion system within the intestines of mice. Furthermore, a single prophylactic dose of a variant of PROT₃EcT that secretes a tumor necrosis factor-alpha (TNF-α)-neutralizing Nb is sufficient to ablate pro-inflammatory TNF levels and prevent the development of injury and inflammation in a chemically induced model of colitis. This work lays the foundation for developing PROT₃EcT as a platform for the treatment of gastrointestinal-based diseases.

Keywords: TNF; colitis; designer probiotic; engineered E. coli; in situ drug delivery; inflammatory bowel disease; live biotherapeutic; nanobody; type III secretion.

MeSH terms

Animals
Colitis* / chemically induced
Colitis* / therapy
Escherichia coli
Mice
Single-Domain Antibodies*
Tumor Necrosis Factor-alpha / metabolism

Substances

Single-Domain Antibodies
Tumor Necrosis Factor-alpha

Abstract

MeSH terms

Substances

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