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Upregulation of virulence genes promotes Vibrio cholerae biofilm hyperinfectivity

Gallego-Hernandez, A. L. and DePas, W. H. and Park, J. H. and Teschler, J. K. and Hartmann, R. and Jeckel, H. and Drescher, K. and Beyhan, S. and Newman, D. K. and Yildiz, F. H. (2020) Upregulation of virulence genes promotes Vibrio cholerae biofilm hyperinfectivity. Proceedings of the National Academy of Sciences of the United States of America, 117 (20). pp. 11010-11017. ISSN 0027-8424. PMCID PMC7245069. doi:10.1073/pnas.1916571117.

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Vibrio cholerae remains a major global health threat, disproportionately impacting parts of the world without adequate infrastructure and sanitation resources. In aquatic environments, V. cholerae exists both as planktonic cells and as biofilms, which are held together by an extracellular matrix. V. cholerae biofilms have been shown to be hyperinfective, but the mechanism of hyperinfectivity is unclear. Here we show that biofilm-grown cells, irrespective of the surfaces on which they are formed, are able to markedly outcompete planktonic-grown cells in the infant mouse. Using an imaging technique designed to render intestinal tissue optically transparent and preserve the spatial integrity of infected intestines, we reveal and compare three-dimensional V. cholerae colonization patterns of planktonic-grown and biofilm-grown cells. Quantitative image analyses show that V. cholerae colonizes mainly the medial portion of the small intestine and that both the abundance and localization patterns of biofilm-grown cells differ from that of planktonic-grown cells. In vitro biofilm-grown cells activate expression of the virulence cascade, including the toxin coregulated pilus (TCP), and are able to acquire the cholera toxin-carrying CTXФ phage. Overall, virulence factor gene expression is also higher in vivo when infected with biofilm-grown cells, and modulation of their regulation is sufficient to cause the biofilm hyperinfectivity phenotype. Together, these results indicate that the altered biogeography of biofilm-grown cells and their enhanced production of virulence factors in the intestine underpin the biofilm hyperinfectivity phenotype.

Item Type:Article
Related URLs:
URLURL TypeDescription Information CentralArticle
Gallego-Hernandez, A. L.0000-0001-7818-2244
Drescher, K.0000-0002-7340-2444
Newman, D. K.0000-0003-1647-1918
Additional Information:© 2020 National Academy of Sciences. Published under the PNAS license. Edited by John J. Mekalanos, Harvard University, Boston, MA, and approved March 11, 2020 (received for review September 23, 2019). PNAS first published April 30, 2020. We thank Benjamin Abrams, University of California, Santa Cruz (UCSC) Life Sciences Microscopy Center, for technical support during confocal imaging, which is supported by grant S10 OD023528 (to F.H.Y.) from the National Institutes of Health. We thank Ron Taylor and Karen Skorupski for the TcpA antibody. We thank Adam Alpine for his assistance with Fig. 2 A and B. We thank Molecular Technologies (Caltech) for assistance with HCR probe design. We would also like to acknowledge the members of the F.H.Y. laboratory and Karla J. F. Satchell for their useful input and contributions. This work was supported by NIH grants RO1 AI102584, RO1 AI114261, and RO1 AI055987 (to F.H.Y.); NIH grants 5R01HL117328-03 and 1R01AI127850-01A1 (to D.K.N.); and the DEPAS17F0 fellowship from the Cystic Fibrosis Foundation (to W.H.D.), as well as the European Research Council StG-716734, the Human Frontier Science Program CDA00084/2015-C, and the Deutsche Forschungsgemeinschaft SFB987 (to K.D.). A.L.G.-H. was supported by the University of California Institute for Mexico and the United States (UC MEXUS) and El Consejo Nacional de Ciencia y Tecnología (CONACYT) Postdoctoral Research fellowship. Data Availability: RNA-seq data are available through NCBI GEO with series number GSE135887. Author contributions: A.L.G.-H., W.H.D., J.H.P., J.K.T., K.D., D.K.N., and F.H.Y. designed research; A.L.G.-H., W.H.D., J.H.P., and J.K.T. performed research; R.H. contributed new reagents/analytic tools; A.L.G.-H., W.H.D., J.H.P., J.K.T., R.H., H.J., K.D., S.B., D.K.N., and F.H.Y. analyzed data; and A.L.G.-H., W.H.D., J.H.P., J.K.T., K.D., D.K.N., and F.H.Y. wrote the paper. The authors declare no competing interest. This article is a PNAS Direct Submission. Data deposition: RNA-seq data are available through National Center for Biotechnology Information (NCBI) Gene Expression Omnibus (GEO) (series number GSE135887). This article contains supporting information online at
Funding AgencyGrant Number
NIHS10 OD023528
NIHRO1 AI102584
NIHRO1 AI114261
NIHRO1 AI055987
Cystic Fibrosis FoundationDEPAS17F0
European Research Council (ERC)716734
Human Frontier Science ProgramCDA00084/2015-C
Deutsche Forschungsgemeinschaft (DFG)SFB987
University of California Institute for Mexico and the United States (UC MEXUS)UNSPECIFIED
Consejo Nacional de Ciencia y Tecnología (CONACYT)UNSPECIFIED
Subject Keywords:Vibrio cholerae; hyperinfectivity; biofilm; gastrointestinal infection
Issue or Number:20
PubMed Central ID:PMC7245069
Record Number:CaltechAUTHORS:20200504-083728290
Persistent URL:
Official Citation:Upregulation of virulence genes promotes Vibrio cholerae biofilm hyperinfectivity. A. L. Gallego-Hernandez, W. H. DePas, J. H. Park, J. K. Teschler, R. Hartmann, H. Jeckel, K. Drescher, S. Beyhan, D. K. Newman, F. H. Yildiz. Proceedings of the National Academy of Sciences May 2020, 117 (20) 11010-11017; DOI: 10.1073/pnas.1916571117
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:102971
Deposited By: Tony Diaz
Deposited On:04 May 2020 16:04
Last Modified:16 Nov 2021 18:17

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