TY - JOUR
T1 - An engineered synthetic biologic protects against Clostridium difficile infection
AU - Vedantam, Gayatri
AU - Kochanowsky, Joshua
AU - Lindsey, Jason
AU - Mallozzi, Michael
AU - Roxas, Jennifer Lising
AU - Adamson, Chelsea
AU - Anwar, Farhan
AU - Clark, Andrew
AU - Claus-Walker, Rachel
AU - Mansoor, Asad
AU - McQuade, Rebecca
AU - Monasky, Ross Calvin
AU - Ramamurthy, Shylaja
AU - Roxas, Bryan
AU - Viswanathan, V. K.
N1 - Funding Information:
Special thanks to Dr. Todd Klaenhammer for the kind gift of the pTRK848 plasmid. Many thanks to all the current and former members of the Vedantam and Viswanathan laboratories, specifically Michele Chu and Andrea Treptow. The help and support of the University of Arizona Cancer Center ARL Cytometry Core Facility (Grant-CA 023074) is gratefully acknowledged. We also thank Dr. Tod McCauley for helpful discussions. This research project was supported by the National Institutes of Health (GV; AI121590). Work in the Vedantam laboratory is also supported by the US Department of Veterans Affairs (1I01BX001183-01) and the USDA CSREES Hatch Program (ARZT-570410-A-02-139; GV). The pilot piglet studies reported herein were supported by an Asset Development Award from Tech Launch Arizona.
Publisher Copyright:
© 2018 Vedantam, Kochanowsky, Lindsey, Mallozzi, Roxas, Adamson, Anwar, Clark, Claus-Walker, Mansoor, McQuade, Monasky, Ramamurthy, Roxas and Viswanathan.
PY - 2018/9/5
Y1 - 2018/9/5
N2 - Morbidity 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 codon-optimized 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, fixed-dose 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.
AB - Morbidity 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 codon-optimized 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, fixed-dose 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.
KW - Clostridium difficile
KW - Infectious diarrhea
KW - Lactobacillus
KW - Surface layer protein
KW - Synthetic biology
UR - http://www.scopus.com/inward/record.url?scp=85053054338&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85053054338&partnerID=8YFLogxK
U2 - 10.3389/fmicb.2018.02080
DO - 10.3389/fmicb.2018.02080
M3 - Article
C2 - 30233548
AN - SCOPUS:85053054338
SN - 1664-302X
VL - 9
JO - Frontiers in Microbiology
JF - Frontiers in Microbiology
IS - SEP
M1 - 2080
ER -