Tags

Type your tag names separated by a space and hit enter

CpxA Phosphatase Inhibitor Activates CpxRA and Is a Potential Treatment for Uropathogenic Escherichia coli in a Murine Model of Infection.
Microbiol Spectr. 2022 04 27; 10(2):e0243021.MS

Abstract

CpxRA is an envelope stress response system that is highly conserved in the Enterobacteriaceae. CpxA has kinase activity for CpxR and phosphatase activity for phospho-CpxR (CpxR-P), a transcription factor. In response to membrane stress, CpxR-P is produced and upregulates genes involved in membrane repair and downregulates genes that encode virulence factors that are trafficked across the cell membrane. Mutants that constitutively activate CpxRA in Salmonella enterica serovar Typhimurium and in uropathogenic Escherichia coli (UPEC) are attenuated in murine models. We hypothesized that pharmacologic activation of CpxR could serve as an antimicrobial/antivirulence strategy and recently showed that 2,3,4,9-tetrahydro-1H-carbazol-1-amines activate the CpxRA system by inhibiting CpxA phosphatase activity. Here, we tested the ability of a series of three CpxRA-activating compounds with increasing potency to clear UPEC stain CFT073 in a murine urinary tract infection model. We show that these compounds are well tolerated and achieve sufficient levels to activate CpxR in the kidneys, bladder, and urine. Although the first two compounds were ineffective in promoting clearance of CFT073 in the murine model, the most potent derivative, compound 26, significantly reduced bacterial recovery in the urine and trended toward reducing bacterial recovery in the bladder and kidneys, with efficacy similar to ciprofloxacin. Treatment of CFT073 cultured in human urine with compound 26 fostered accumulation of CpxR-P and decreased the expression of proteins involved in siderophore biosynthesis and binding, heme degradation, and flagellar movement. These studies suggest that chemical activation of CpxRA may present a viable strategy for treating infections due to UPEC. IMPORTANCE The increasing prevalence of urinary tract infections (UTIs) due to antibiotic-resistant uropathogenic Escherichia coli (UPEC) is a major public health concern. Bacteria contain proteins that sense their environment and have no human homologs and, thus, are attractive drug targets. CpxRA is a conserved sensing system whose function is to reduce stress in the bacterial cell membrane; activation of CpxRA reduces the expression of virulence determinants, which must cross the cell membrane to reach the bacterial surface. We previously identified a class of compounds that activate CpxRA. We show in a mouse UTI model that our most potent compound significantly reduced recovery of UPEC in the urine, trended toward reducing bacterial recovery in the bladder and kidneys, did not kill UPEC, and downregulated multiple proteins involved in UPEC virulence. Since these compounds do not act by a killing mechanism, they have potential to treat UTIs caused by antibiotic-resistant bacteria.

Authors+Show Affiliations

Department of Microbiology and Immunology, Indiana University School of Medicine, Indianapolis, Indiana, USA.Department of Microbiology and Immunology, University of Michigan Medical Schoolgrid.471406.0, Ann Arbor, Michigan, USA.Department of Microbiology and Immunology, Indiana University School of Medicine, Indianapolis, Indiana, USA.Department of Microbiology and Immunology, Indiana University School of Medicine, Indianapolis, Indiana, USA.Department of Chemistry and Biochemistry, University of Oklahoma, Norman, Oklahoma, USA.Department of Biostatistics, Indiana University School of Medicine, Indianapolis, Indiana, USA. Department of Biostatistics, Richard M. Fairbanks School of Public Health, Indiana University, Indianapolis, Indiana, USA.Department of Chemistry and Biochemistry, University of Oklahoma, Norman, Oklahoma, USA. Mass Spectrometry, Proteomics and Metabolomics Core Facility, Stephenson Life Sciences Research Center, University of Oklahoma, Norman, Oklahoma, USA.Department of Medicinal Chemistry, University of Minnesota, Minneapolis, Minnesota, USA.Department of Microbiology and Immunology, University of Michigan Medical Schoolgrid.471406.0, Ann Arbor, Michigan, USA.Department of Microbiology and Immunology, Indiana University School of Medicine, Indianapolis, Indiana, USA. Department of Medicine, Indiana University School of Medicine, Indianapolis, Indiana, USA. Department of Pathology and Laboratory Medicine, Indiana University School of Medicine, Indianapolis, Indiana, USA.

Pub Type(s)

Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

35297652

Citation

Fortney, Kate R., et al. "CpxA Phosphatase Inhibitor Activates CpxRA and Is a Potential Treatment for Uropathogenic Escherichia Coli in a Murine Model of Infection." Microbiology Spectrum, vol. 10, no. 2, 2022, pp. e0243021.
Fortney KR, Smith SN, van Rensburg JJ, et al. CpxA Phosphatase Inhibitor Activates CpxRA and Is a Potential Treatment for Uropathogenic Escherichia coli in a Murine Model of Infection. Microbiol Spectr. 2022;10(2):e0243021.
Fortney, K. R., Smith, S. N., van Rensburg, J. J., Brothwell, J. A., Gardner, J. J., Katz, B. P., Ahsan, N., Duerfeldt, A. S., Mobley, H. L. T., & Spinola, S. M. (2022). CpxA Phosphatase Inhibitor Activates CpxRA and Is a Potential Treatment for Uropathogenic Escherichia coli in a Murine Model of Infection. Microbiology Spectrum, 10(2), e0243021. https://doi.org/10.1128/spectrum.02430-21
Fortney KR, et al. CpxA Phosphatase Inhibitor Activates CpxRA and Is a Potential Treatment for Uropathogenic Escherichia Coli in a Murine Model of Infection. Microbiol Spectr. 2022 04 27;10(2):e0243021. PubMed PMID: 35297652.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - CpxA Phosphatase Inhibitor Activates CpxRA and Is a Potential Treatment for Uropathogenic Escherichia coli in a Murine Model of Infection. AU - Fortney,Kate R, AU - Smith,Sara N, AU - van Rensburg,Julia J, AU - Brothwell,Julie A, AU - Gardner,Jessi J, AU - Katz,Barry P, AU - Ahsan,Nagib, AU - Duerfeldt,Adam S, AU - Mobley,Harry L T, AU - Spinola,Stanley M, Y1 - 2022/03/17/ PY - 2022/3/18/pubmed PY - 2022/4/30/medline PY - 2022/3/17/entrez KW - CpxRA KW - Escherichia coli KW - UPEC KW - phosphatase inhibitor KW - treatment SP - e0243021 EP - e0243021 JF - Microbiology spectrum JO - Microbiol Spectr VL - 10 IS - 2 N2 - CpxRA is an envelope stress response system that is highly conserved in the Enterobacteriaceae. CpxA has kinase activity for CpxR and phosphatase activity for phospho-CpxR (CpxR-P), a transcription factor. In response to membrane stress, CpxR-P is produced and upregulates genes involved in membrane repair and downregulates genes that encode virulence factors that are trafficked across the cell membrane. Mutants that constitutively activate CpxRA in Salmonella enterica serovar Typhimurium and in uropathogenic Escherichia coli (UPEC) are attenuated in murine models. We hypothesized that pharmacologic activation of CpxR could serve as an antimicrobial/antivirulence strategy and recently showed that 2,3,4,9-tetrahydro-1H-carbazol-1-amines activate the CpxRA system by inhibiting CpxA phosphatase activity. Here, we tested the ability of a series of three CpxRA-activating compounds with increasing potency to clear UPEC stain CFT073 in a murine urinary tract infection model. We show that these compounds are well tolerated and achieve sufficient levels to activate CpxR in the kidneys, bladder, and urine. Although the first two compounds were ineffective in promoting clearance of CFT073 in the murine model, the most potent derivative, compound 26, significantly reduced bacterial recovery in the urine and trended toward reducing bacterial recovery in the bladder and kidneys, with efficacy similar to ciprofloxacin. Treatment of CFT073 cultured in human urine with compound 26 fostered accumulation of CpxR-P and decreased the expression of proteins involved in siderophore biosynthesis and binding, heme degradation, and flagellar movement. These studies suggest that chemical activation of CpxRA may present a viable strategy for treating infections due to UPEC. IMPORTANCE The increasing prevalence of urinary tract infections (UTIs) due to antibiotic-resistant uropathogenic Escherichia coli (UPEC) is a major public health concern. Bacteria contain proteins that sense their environment and have no human homologs and, thus, are attractive drug targets. CpxRA is a conserved sensing system whose function is to reduce stress in the bacterial cell membrane; activation of CpxRA reduces the expression of virulence determinants, which must cross the cell membrane to reach the bacterial surface. We previously identified a class of compounds that activate CpxRA. We show in a mouse UTI model that our most potent compound significantly reduced recovery of UPEC in the urine, trended toward reducing bacterial recovery in the bladder and kidneys, did not kill UPEC, and downregulated multiple proteins involved in UPEC virulence. Since these compounds do not act by a killing mechanism, they have potential to treat UTIs caused by antibiotic-resistant bacteria. SN - 2165-0497 UR - https://www.unboundmedicine.com/medline/citation/35297652/CpxA_Phosphatase_Inhibitor_Activates_CpxRA_and_Is_a_Potential_Treatment_for_Uropathogenic_Escherichia_coli_in_a_Murine_Model_of_Infection_ DB - PRIME DP - Unbound Medicine ER -