An Engineered Synthetic Biologic Protects Against Clostridium difficile Infection
Roxas, Jennifer Lising
Monasky, Ross Calvin
Viswanathan, V. K.
AffiliationUniv Arizona, Sch Anim & Comparat Biomed Sci
Univ Arizona, Dept Immunobiol
Univ Arizona, Inst Collaborat Res Bio5
surface layer protein
MetadataShow full item record
PublisherFRONTIERS MEDIA SA
CitationVedantam G, Kochanowsky J, Lindsey J, Mallozzi M, Roxas JL, Adamson C, Anwar F, Clark A, Claus-Walker R, Mansoor A, McQuade R, Monasky R, Ramamurthy S, Roxas B and Viswanathan VK (2018) An Engineered Synthetic Biologic Protects Against Clostridium difficile Infection. Front. Microbiol. 9:2080. doi: 10.3389/fmicb.2018.02080
JournalFRONTIERS IN MICROBIOLOGY
Rights© 2018 Vedantam, Kochanowsky, Lindsey, Mallozzi, Roxas, Adamson, Anwar, Clark, Claus-Walker, Mansoor, McQuade, Monasky, Ramamurthy, Roxas and Viswanathan. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY).
Collection InformationThis item from the UA Faculty Publications collection is made available by the University of Arizona with support from the University of Arizona Libraries. If you have questions, please contact us at firstname.lastname@example.org.
AbstractMorbidity and mortality attributed to Clostridium difficile infection (CDI) have increased over the past 20 years. Currently, antibiotics are the only US FDA-approved treatment for primary C. difficile infection, and these are, ironically, associated with disease relapse and the threat of burgeoning drug resistance. We previously showed that non-toxin virulence factors play key roles in CDI, and that colonization factors are critical for disease. Specifically, a C. difficile adhesin, Surface Layer Protein A (SlpA) is a major contributor to host cell attachment. In this work, we engineered Syn-LAB 2.0 and Syn-LAB 2.1, two synthetic biologic agents derived from lactic acid bacteria, to stably and constitutively express a host-cell binding fragment of the C. difficile adhesin SlpA on their cell-surface. Both agents harbor conditional suicide plasmids expressing a codonoptimized chimera of the lactic acid bacterium's cell-wall anchoring surface-protein domain, fused to the conserved, highly adherent, host-cell-binding domain of C. difficile SlpA. Both agents also incorporate engineered biocontrol, obviating the need for any antibiotic selection. Syn-LAB 2.0 and Syn-LAB 2.1 possess positive biophysical and in vivo properties compared with their parental antecedents in that they robustly and constitutively display the SlpA chimera on their cell surface, potentiate human intestinal epithelial barrier function in vitro, are safe, tolerable and palatable to Golden Syrian hamsters and neonatal piglets at high daily doses, and are detectable in animal feces within 24 h of dosing, confirming robust colonization. In combination, the engineered strains also delay (in fixed doses) or prevent (when continuously administered) death of infected hamsters upon challenge with high doses of virulent C. difficile. Finally, fixeddose Syn-LAB ameliorates diarrhea in a non-lethal model of neonatal piglet enteritis. Taken together, our findings suggest that the two synthetic biologics may be effectively employed as non-antibiotic interventions for CDI.
NoteOpen access journal.
VersionFinal published version
SponsorsNational Institutes of Health [AI121590]; US Department of Veterans Affairs [1I01BX001183-01]; USDA CSREES Hatch Program [ARZT-570410-A-02-139]; Asset Development Award from Tech Launch Arizona