Characterization of Colibactin-Producing &lt;i&gt;Escherichia coli&lt;/i&gt; Isolated from Japanese Patients with Colorectal Cancer

Yoshikawa, Yuko; Tsunematsu, Yuta; Matsuzaki, Nobuo; Hirayama, Yoshitaka; Higashiguchi, Fumi; Sato, Michio; Iwashita, Yuji; Miyoshi, Noriyuki; Mutoh, Michihiro; Ishikawa, Hideki; Sugimura, Haruhiko; Wakabayashi, Keiji; Watanabe, Kenji

doi:10.7883/yoken.jjid.2020.066

Cited by 22 publications

(19 citation statements)

References 35 publications

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“…A recent clinical observation study found E. coli HlyA and colibactin to be present in high abundance on colorectal cancer [31]. This observation, together with our experimental association of HlyA to the leaky gut with influx of noxious agents and to the pro-oncogenic PTEN inhibition as well as to the cell separation or even dissemination of colon carcinoma Caco-2 cells, could open new perspectives on the surveillance of these virulence factors in colorectal cancer and metastases.…”

Section: Discussionsupporting

confidence: 67%

Escherichia coli Alpha-Hemolysin HlyA Induces Host Cell Polarity Changes, Epithelial Barrier Dysfunction and Cell Detachment in Human Colon Carcinoma Caco-2 Cell Model via PTEN-Dependent Dysregulation of Cell Junctions

Schulz

Schümann

Schneemann

et al. 2021

Toxins

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Escherichia coli (E. coli) of the B2 phylotype reside in human and animal intestines. The bacteria possess pathogenicity factors such as α-hemolysin (HlyA) that can induce intestinal epithelial leaks. We addressed the questions which host cell processes were dysregulated by E. coli HlyA that can potentiate intestinal diseases. The colon carcinoma cell line Caco-2 was infected by HlyA+ E. coli. Cell polarity regulation was analyzed by live cell imaging for the phosphatidylinositol-4,5-bisphosphate (PIP2) abundance. In Caco-2 monolayers, transepithelial electrical resistance was measured for characterization of barrier function. Cell proliferation and separation were assessed microscopically. Epithelial regulation and cell signaling were analyzed by RNA-Seq and Ingenuity Pathway Analysis (IPA). Our main findings from E. coli HlyA toxinogenicity in the colon carcinoma cell line are that (i) PIP2 at the membrane decrease, (ii) PTEN (phosphatase and tensin homolog) inhibition leads to cell polarity changes, (iii) epithelial leakiness follows these polarity changes by disruption of cell junctions and (iv) epithelial cell detachment increases. HlyA affected pathways, e.g., the PTEN and metastasis signaling, were identified by RNA-Seq bioinformatics calculations in IPA. In conclusion, HlyA affects cell polarity, thereby inducing epithelial barrier dysfunction due to defective tight junctions and focal leak induction as an exemplary mechanism for leaky gut.

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Section: Discussionsupporting

confidence: 67%

Escherichia coli Alpha-Hemolysin HlyA Induces Host Cell Polarity Changes, Epithelial Barrier Dysfunction and Cell Detachment in Human Colon Carcinoma Caco-2 Cell Model via PTEN-Dependent Dysregulation of Cell Junctions

Schulz

Schümann

Schneemann

et al. 2021

Toxins

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“…The aim of this study was to investigate whether prebiotics, inulin and GOS, could modulate the expression and toxicity of the genotoxin colibactin, which has been linked to CRC development. Given the well-established link between diet (26), gut microbiota (27), and CRC a better understanding of how colibactin expression is regulated by different nutrients and supplements is of paramount importance due to the increasing presence of pks + E. coli in healthy individuals in Western countries (5,28) and, more recently, among Malaysian (29), Indian (30), and Japanese populations (31).…”

Section: Discussionmentioning

confidence: 99%

Inulin and Galacto-oligosaccharides Increase the Genotoxic Effect of Colibactin Produced by pks+ Escherichia Coli Strains

Oliero¹,

Calvé²,

Fragoso³

et al. 2020

Preprint

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Background: Colibactin is a genotoxin that induces double-strand DNA breaks and is produced by Escherichia coli strains harboring the pks island. Human and animal studies have shown that colibactin-producing gut bacteria promote carcinogenesis and enhance the progression of colorectal cancer through cellular senescence and chromosomal abnormalities. In this study, we investigated the impact of prebiotics on the genotoxicity of colibactin-producing E. coli strains Nissle 1917 and NC101. Methods: Bacteria were grown in medium supplemented with 20, 30 and 40 mg/mL of prebiotics inulin or galacto-oligosaccharide, and with or without 5 µM, 25 µM and 125 µM of iron sulfate. Colibactin expression was assessed by luciferase reporter assay for the clbA gene, essential for colibactin production, in E. coli Nissle 1917 and by RT-PCR in E. coli NC101. The human epithelial colorectal adenocarcinoma cell line, Caco-2, was used to assess colibactin-induced megalocytosis by methylene blue binding assay and genotoxicity by γ-H2AX immunofluorescence analysis. Results: Inulin and galacto-oligosaccharide enhanced the expression of clbA in pks+ E. coli. However, the addition of 125 µM of iron sulfate inhibited the expression of clbA triggered by oligosaccharides. In the presence of either oligosaccharide, E. coli NC101 increased dysplasia and double-strand DNA breaks in Caco-2 cells compared to untreated cells. Conclusion: Our results suggest that, in vitro, prebiotic oligosaccharides exacerbate DNA damage induced by colibactin-producing bacteria. Further studies are necessary to establish whether these results are reproducible in vivo.

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“…Virulence factor profiling also showed an increased prevalence of the hemolysin system (hly) in pks-positive isolates (Fig. 1), the association of which was indicated previously as a risk factor for colorectal cancer (53). The study was further expanded to screen 4,090 genomes of E. coli obtained from NCBI, out of which the pks island was detected in 507 genomes, in addition to the 23 in-house genomes.…”

Section: Discussionmentioning

confidence: 69%

Evolutionary Dynamics Based on Comparative Genomics of Pathogenic Escherichia coli Lineages Harboring Polyketide Synthase ( pks ) Island

Suresh

Shaik

Baddam

et al. 2021

mBio

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The genotoxin colibactin is a secondary metabolite produced by the polyketide synthase (pks) island harbored by extraintestinal pathogenic E. coli (ExPEC) and other members of the Enterobacteriaceae that has been increasingly reported to have critical implications in human health. The present study entails a high-throughput whole-genome comparison and phylogenetic analysis of such pathogenic E. coli isolates to gain insights into the patterns of distribution, horizontal transmission, and evolution of the island. For the current study, 23 pks-positive ExPEC genomes were newly sequenced, and their virulome and resistome profiles indicated a preponderance of virulence encoding genes and a reduced number of genes for antimicrobial resistance. In addition, 4,090 E. coli genomes from the public domain were also analyzed for large-scale screening for pks-positive genomes, out of which a total of 530 pks-positive genomes were studied to understand the subtype-based distribution pattern(s). The pks island showed a significant association with the B2 phylogroup (82.2%) and a high prevalence in sequence type 73 (ST73; n = 179) and ST95 (n = 110) and the O6:H1 (n = 110) serotype. Maximum-likelihood (ML) phylogeny of the core genome and intergenic regions (IGRs) of the ST95 model data set, which was selected because it had both pks-positive and pks-negative genomes, displayed clustering in relation to their carriage of the pks island. Prevalence patterns of genes encoding RM systems in the pks-positive and pks-negative genomes were also analyzed to determine their potential role in pks island acquisition and the maintenance capability of the genomes. Further, the maximum-likelihood phylogeny based on the core genome and pks island sequences from 247 genomes with an intact pks island demonstrated horizontal gene transfer of the island across sequence types and serotypes, with few exceptions. This study vitally contributes to understanding of the lineages and subtypes that have a higher propensity to harbor the pks island-encoded genotoxin with possible clinical implications. IMPORTANCE Extraintestinal pathologies caused by highly virulent strains of E. coli amount to clinical implications with high morbidity and mortality rates. Pathogenic E. coli strains are evolving with the horizontal acquisition of mobile genetic elements, including pathogenicity islands such as the pks island, which produces the genotoxin colibactin, resulting in severe clinical outcomes, including colorectal cancer progression. The current study encompasses high-throughput comparative genomics and phylogenetic analyses to address the questions pertaining to the acquisition and evolution pattern of the genomic island in different E. coli subtypes. It is crucial to gain insights into the distribution, transfer, and maintenance of pathogenic islands, as they harbor multiple virulence genes involved in pathogenesis and clinical implications of the infection.

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Characterization of Colibactin-Producing <i>Escherichia coli</i> Isolated from Japanese Patients with Colorectal Cancer

Cited by 22 publications

References 35 publications

Escherichia coli Alpha-Hemolysin HlyA Induces Host Cell Polarity Changes, Epithelial Barrier Dysfunction and Cell Detachment in Human Colon Carcinoma Caco-2 Cell Model via PTEN-Dependent Dysregulation of Cell Junctions

Escherichia coli Alpha-Hemolysin HlyA Induces Host Cell Polarity Changes, Epithelial Barrier Dysfunction and Cell Detachment in Human Colon Carcinoma Caco-2 Cell Model via PTEN-Dependent Dysregulation of Cell Junctions

Inulin and Galacto-oligosaccharides Increase the Genotoxic Effect of Colibactin Produced by pks+ Escherichia Coli Strains

Evolutionary Dynamics Based on Comparative Genomics of Pathogenic Escherichia coli Lineages Harboring Polyketide Synthase ( pks ) Island

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