title: Engineered Phagemids for Nonlytic, Targeted Antibacterial Therapies.
abstract: The increasing incidence of antibiotic-resistant bacterial infections is creating a global public health threat. Because conventional antibiotic drug discovery has failed to keep pace with the rise of resistance, a growing need exists to develop novel antibacterial methodologies. Replication-competent bacteriophages have been utilized in a limited fashion to treat bacterial infections. However, this approach can result in the release of harmful endotoxins, leading to untoward side effects. Here, we engineer bacterial phagemids to express antimicrobial peptides (AMPs) and protein toxins that disrupt intracellular processes, leading to rapid, nonlytic bacterial death. We show that this approach is highly modular, enabling one to readily alter the number and type of AMPs and toxins encoded by the phagemids. Furthermore, we demonstrate the effectiveness of engineered phagemids in an in vivo murine peritonitis infection model. This work shows that targeted, engineered phagemid therapy can serve as a viable, nonantibiotic means to treat bacterial infections, while avoiding the health issues inherent to lytic and replicative bacteriophage use.
authors.affiliation: †Institute for Medical Engineering and Science, Department of Biological Engineering, and Synthetic Biology Center, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, United States.|‡Harvard-MIT Program in Health Sciences and Technology, Cambridge, Massachusetts 02139, United States.|∥Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, Massachusetts 02115, United States.|⊥Department of Molecular and Translational Medicine, Boston University, Boston, Massachusetts 02215, United States.
authors.forename: Russell J
authors.initials: RJ
authors.lastname: Krom
authors.affiliation: †Institute for Medical Engineering and Science, Department of Biological Engineering, and Synthetic Biology Center, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, United States.|§Broad Institute of MIT and Harvard, Cambridge, Massachusetts 02142, United States.|∥Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, Massachusetts 02115, United States.
authors.forename: Prerna
authors.initials: P
authors.lastname: Bhargava
authors.affiliation: †Institute for Medical Engineering and Science, Department of Biological Engineering, and Synthetic Biology Center, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, United States.|§Broad Institute of MIT and Harvard, Cambridge, Massachusetts 02142, United States.|∥Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, Massachusetts 02115, United States.|#Division of Infectious Diseases, Massachusetts General Hospital, Boston, Massachusetts 02114, United States.
authors.forename: Michael A
authors.initials: MA
authors.lastname: Lobritz
authors.affiliation: †Institute for Medical Engineering and Science, Department of Biological Engineering, and Synthetic Biology Center, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, United States.|‡Harvard-MIT Program in Health Sciences and Technology, Cambridge, Massachusetts 02139, United States.|§Broad Institute of MIT and Harvard, Cambridge, Massachusetts 02142, United States.|∥Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, Massachusetts 02115, United States.
authors.forename: James J
authors.initials: JJ
authors.lastname: Collins
mesh_terms: D000818:Animals; D000900:Anti-Bacterial Agents / Q000378:metabolism*; D023181:Antimicrobial Cationic Peptides / Q000235:genetics*; D001435:Bacteriophages / Q000235:genetics*; D004926:Escherichia coli / Q000502:physiology; D004927:Escherichia coli Infections / Q000150:complications; D005818:Genetic Engineering; D015316:Genetic Therapy; D006801:Humans; D051379:Mice; D008810:Mice, Inbred C57BL; D010538:Peritonitis / Q000235:genetics / Q000382:microbiology / Q000628:therapy*; D010957:Plasmids / Q000235:genetics / Q000627:therapeutic use*; D014118:Toxins, Biological / Q000235:genetics*
chemical_list: D000900:Anti-Bacterial Agents; D023181:Antimicrobial Cationic Peptides; D014118:Toxins, Biological
keywords: Antimicrobial peptides; bacterial toxins; bacteriophage resistance; phagemids; synthetic gene networks; targeted bacterial killing