Lifting the mask: Identification of new small molecule inhibitors of uropathogenic Escherichia coli group 2 capsule biogenesis

Carlos C. Goller, Mehreen Arshad, James W. Noah, Subramaniam Ananthan, Carrie W. Evans, N. Miranda Nebane, Lynn Rasmussen, Melinda Sosa, Nichole A. Tower, E. Lucile White, Benjamin Neuenswander, Patrick Porubsky, Brooks E. Maki, Steven A. Rogers, Frank Schoenen, Patrick C. Seed

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Uropathogenic Escherichia coli (UPEC) is the leading cause of community-acquired urinary tract infections (UTIs), with over 100 million UTIs occurring annually throughout the world. Increasing antimicrobial resistance among UPEC limits ambulatory care options, delays effective treatment, and may increase overall morbidity and mortality from complications such as urosepsis. The polysaccharide capsules of UPEC are an attractive target a therapeutic, based on their importance in defense against the host immune responses; however, the large number of antigenic types has limited their incorporation into vaccine development. The objective of this study was to identify small-molecule inhibitors of UPEC capsule biogenesis. A large-scale screening effort entailing 338,740 compounds was conducted in a cell-based, phenotypic screen for inhibition of capsule biogenesis in UPEC. The primary and concentration-response assays yielded 29 putative inhibitors of capsule biogenesis, of which 6 were selected for further studies. Secondary confirmatory assays identified two highly active agents, named DU003 and DU011, with 50% inhibitory concentrations of 1.0 mM and 0.69 mM, respectively. Confirmatory assays for capsular antigen and biochemical measurement of capsular sugars verified the inhibitory action of both compounds and demonstrated minimal toxicity and off-target effects. Serum sensitivity assays demonstrated that both compounds produced significant bacterial death upon exposure to active human serum. DU011 administration in mice provided near complete protection against a lethal systemic infection with the prototypic UPEC K1 isolate UTI89. This work has provided a conceptually new class of molecules to combat UPEC infection, and future studies will establish the molecular basis for their action along with efficacy in UTI and other UPEC infections.

Original languageEnglish (US)
Article numbere96054
JournalPLoS One
Volume9
Issue number7
DOIs
StatePublished - Jul 1 2014

Fingerprint

uropathogenic Escherichia coli
Uropathogenic Escherichia coli
Masks
Escherichia coli
Capsules
Molecules
urinary tract diseases
Assays
Urinary Tract Infections
Escherichia coli Infections
assays
infection
Community-Acquired Infections
biogenesis
vaccine development
Ambulatory Care
Serum
Sugars
antibiotic resistance
Inhibitory Concentration 50

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

Goller, Carlos C. ; Arshad, Mehreen ; Noah, James W. ; Ananthan, Subramaniam ; Evans, Carrie W. ; Nebane, N. Miranda ; Rasmussen, Lynn ; Sosa, Melinda ; Tower, Nichole A. ; White, E. Lucile ; Neuenswander, Benjamin ; Porubsky, Patrick ; Maki, Brooks E. ; Rogers, Steven A. ; Schoenen, Frank ; Seed, Patrick C. / Lifting the mask : Identification of new small molecule inhibitors of uropathogenic Escherichia coli group 2 capsule biogenesis. In: PLoS One. 2014 ; Vol. 9, No. 7.
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abstract = "Uropathogenic Escherichia coli (UPEC) is the leading cause of community-acquired urinary tract infections (UTIs), with over 100 million UTIs occurring annually throughout the world. Increasing antimicrobial resistance among UPEC limits ambulatory care options, delays effective treatment, and may increase overall morbidity and mortality from complications such as urosepsis. The polysaccharide capsules of UPEC are an attractive target a therapeutic, based on their importance in defense against the host immune responses; however, the large number of antigenic types has limited their incorporation into vaccine development. The objective of this study was to identify small-molecule inhibitors of UPEC capsule biogenesis. A large-scale screening effort entailing 338,740 compounds was conducted in a cell-based, phenotypic screen for inhibition of capsule biogenesis in UPEC. The primary and concentration-response assays yielded 29 putative inhibitors of capsule biogenesis, of which 6 were selected for further studies. Secondary confirmatory assays identified two highly active agents, named DU003 and DU011, with 50{\%} inhibitory concentrations of 1.0 mM and 0.69 mM, respectively. Confirmatory assays for capsular antigen and biochemical measurement of capsular sugars verified the inhibitory action of both compounds and demonstrated minimal toxicity and off-target effects. Serum sensitivity assays demonstrated that both compounds produced significant bacterial death upon exposure to active human serum. DU011 administration in mice provided near complete protection against a lethal systemic infection with the prototypic UPEC K1 isolate UTI89. This work has provided a conceptually new class of molecules to combat UPEC infection, and future studies will establish the molecular basis for their action along with efficacy in UTI and other UPEC infections.",
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Goller, CC, Arshad, M, Noah, JW, Ananthan, S, Evans, CW, Nebane, NM, Rasmussen, L, Sosa, M, Tower, NA, White, EL, Neuenswander, B, Porubsky, P, Maki, BE, Rogers, SA, Schoenen, F & Seed, PC 2014, 'Lifting the mask: Identification of new small molecule inhibitors of uropathogenic Escherichia coli group 2 capsule biogenesis', PLoS One, vol. 9, no. 7, e96054. https://doi.org/10.1371/journal.pone.0096054

Lifting the mask : Identification of new small molecule inhibitors of uropathogenic Escherichia coli group 2 capsule biogenesis. / Goller, Carlos C.; Arshad, Mehreen; Noah, James W.; Ananthan, Subramaniam; Evans, Carrie W.; Nebane, N. Miranda; Rasmussen, Lynn; Sosa, Melinda; Tower, Nichole A.; White, E. Lucile; Neuenswander, Benjamin; Porubsky, Patrick; Maki, Brooks E.; Rogers, Steven A.; Schoenen, Frank; Seed, Patrick C.

In: PLoS One, Vol. 9, No. 7, e96054, 01.07.2014.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Lifting the mask

T2 - Identification of new small molecule inhibitors of uropathogenic Escherichia coli group 2 capsule biogenesis

AU - Goller, Carlos C.

AU - Arshad, Mehreen

AU - Noah, James W.

AU - Ananthan, Subramaniam

AU - Evans, Carrie W.

AU - Nebane, N. Miranda

AU - Rasmussen, Lynn

AU - Sosa, Melinda

AU - Tower, Nichole A.

AU - White, E. Lucile

AU - Neuenswander, Benjamin

AU - Porubsky, Patrick

AU - Maki, Brooks E.

AU - Rogers, Steven A.

AU - Schoenen, Frank

AU - Seed, Patrick C.

PY - 2014/7/1

Y1 - 2014/7/1

N2 - Uropathogenic Escherichia coli (UPEC) is the leading cause of community-acquired urinary tract infections (UTIs), with over 100 million UTIs occurring annually throughout the world. Increasing antimicrobial resistance among UPEC limits ambulatory care options, delays effective treatment, and may increase overall morbidity and mortality from complications such as urosepsis. The polysaccharide capsules of UPEC are an attractive target a therapeutic, based on their importance in defense against the host immune responses; however, the large number of antigenic types has limited their incorporation into vaccine development. The objective of this study was to identify small-molecule inhibitors of UPEC capsule biogenesis. A large-scale screening effort entailing 338,740 compounds was conducted in a cell-based, phenotypic screen for inhibition of capsule biogenesis in UPEC. The primary and concentration-response assays yielded 29 putative inhibitors of capsule biogenesis, of which 6 were selected for further studies. Secondary confirmatory assays identified two highly active agents, named DU003 and DU011, with 50% inhibitory concentrations of 1.0 mM and 0.69 mM, respectively. Confirmatory assays for capsular antigen and biochemical measurement of capsular sugars verified the inhibitory action of both compounds and demonstrated minimal toxicity and off-target effects. Serum sensitivity assays demonstrated that both compounds produced significant bacterial death upon exposure to active human serum. DU011 administration in mice provided near complete protection against a lethal systemic infection with the prototypic UPEC K1 isolate UTI89. This work has provided a conceptually new class of molecules to combat UPEC infection, and future studies will establish the molecular basis for their action along with efficacy in UTI and other UPEC infections.

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