Isolation and Functional Characterization of Fusobacterium nucleatum Bacteriophage

Kabwe, Mwila; Brown, Teagan L.; Ku, Heng; Dashper, Stuart G.; Tucci, Joseph

doi:10.1007/978-1-0716-1518-8_4

Cited by 4 publications

(3 citation statements)

References 42 publications

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“…Last but not least, bacteriophages may provide an accurate means for targeting intratumoral "bad" microbiota, which have an increased abundance in tumor tissue (121). Currently, multiple bacteriophages targeting F. nucleatum have been isolated (122,123), making them a promising adjunct to augment existing therapies against breast cancers. Bacteriophages are highly species-specific; for example, five subspecies of F. nucleatum have been identified: animalis, fusiforme, nucleatum, polymorphum, and vincentii (124).…”

Section: Could Intratumoral Microbiota Serve As a Biomarker Or Treatm...mentioning

confidence: 99%

Intratumoral Microbiota: Unraveling their Oncogenic Impact on Cancer Progression With Focus on Breast Cancer Therapeutic Outcomes

CHANG,

PEKKLE LAM

2024

Anticancer Res

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The gut microbiota has been implicated in many cancers through the secretion of blood-traveling metabolites or activation of oncogenic signaling. Currently, specific microbial signatures have been detected in the human breast, which are different from other microbial-rich compartments, such as the intestine and skin. Changes in the breast microbiome profile have been shown to positively or negatively correlate with breast cancer development, progression, and therapeutic outcomes. However, studies regarding the role and underlying mechanism of intratumoral microbiota in breast cancer have remained concealed. This review aimed to provide an overview of the role of the intratumoral microbiome in tumorigenesis and tumor progression, and how these intratumoral microbiota affect breast cancer. We also discuss the potential of using the intratumoral microbiome as a biomarker or treatment alternative in breast cancers.

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Section: Could Intratumoral Microbiota Serve As a Biomarker Or Treatm...mentioning

confidence: 99%

Intratumoral Microbiota: Unraveling their Oncogenic Impact on Cancer Progression With Focus on Breast Cancer Therapeutic Outcomes

CHANG,

PEKKLE LAM

2024

Anticancer Res

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“…Wastewater samples from two different sites (Gisborne and Woodend, VIC, Australia) were shown to produce plaques on KLEB009. Isolated bacteriophages vB_KpnP_KPN7 (KPN7) and vB_KpnM_KPN8 (KPN8), were then purified from their single plaques and concentrated as described previously [29]. This provided a "crude" purification KPN7 and KPN8.…”

Section: Bacteriophage Isolation and Purificationmentioning

confidence: 99%

“…Bacteriophage DNA was extracted as previously described [29]. Ten mL PBS (pH 7.4) suspensions of Klebsiella bacteriophages KPN7 and KPN8 at concentrations of 1 × 10 8 PFU mL −1 were treated with 5 mmol/L of MgCl 2 (Sigma-Aldrich ® , Macquarie Park, Australia), 10 µg/mL RNAse A, and 10 µg/mL DNAse I (Promega, Alexandria, Australia) for 30 min at room temperature.…”

Section: Bacteriophage Whole Genome Sequencing and In Silico Analysismentioning

confidence: 99%

Effects of Klebsiella pneumoniae Bacteriophages on IRAK3 Knockdown/Knockout THP-1 Monocyte Cell Lines

Schubert

Kabwe

et al. 2022

Viruses

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Bacterial sepsis characterised by an immunosuppressive and cytokine storm state is a challenge to treat clinically. While conventional antibiotics have been associated with exacerbating the cytokine storm, the role that bacteriophages may play in immune modulation of sepsis remains unclear. Bacteriophages are bacterial viruses that have the capacity to lyse specific bacteria and hence provide a natural alternative to antibiotics. K. pneumoniae is known to cause sepsis in humans, and in this study we isolated two lytic bacteriophages against this pathogen, one of which was a novel jumbo bacteriophage. We employed THP-1 monocyte cell lines, with different functional phenotypes for the interleukin-1 receptor associated kinase 3 (IRAK3- a cytoplasmic homeostatic mediator and prognostic marker of inflammation), to evaluate the role of the K. pneumoniae bacteriophages in modulating the immune response in-vitro. We showed for the first time that bacteriophages did not stimulate excessive production of tumour necrosis factor alpha, or interleukin-6, in THP-1 monocyte cell lines which displayed varying levels of IRAK3 expression.

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Evaluation of effectiveness of bacteriophage purification methods

Binte Mohamed Yakob Adil,

Tucci,

Irving

et al. 2024

Virol J

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The use of bacteriophages for therapy has increased over the last decade. While there is need for clear regulatory pathways for bacteriophage approval for mainstream use in clinical practice, practitioners and patients have been able to access bacteriophage therapy under compassionate grounds and through magistral preparations. However, there is currently no standard for purifying these bacteriophages to ensure safety, and good manufacturing practice certification may not be achieved in these emergency uses. In this study, we employed an Interleukin Receptor Associated Kinase (IRAK) 3 knockout monocyte-based assay to evaluate the endotoxin removal efficacy of three common bacteriophage purification methods: Triton X-100 exposure, CsCl density gradient ultracentrifugation, and Pierce™ High-Capacity Endotoxin Removal Resin spin columns. In our experiments we tested these purification methods on three different bacteriophage morphotypes: siphovirus, podovirus and myovirus. We showed that the lowest endotoxin levels and immune responses were achieved when purifying bacteriophages with Triton-X treatment. The results from purifying with CsCl density gradient ultracentrifugation were comparable, and these were both significantly better than purification with Pierce™ High-Capacity Endotoxin Removal Resin spin columns. We also showed that Triton X-100 purification resulted in the lowest loss of bacteriophage titres. Finally, of the bacteriophages tested here, it did not appear that virus morphology affected efficacy of endotoxin removal.

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Isolation and Functional Characterization of Fusobacterium nucleatum Bacteriophage

Cited by 4 publications

References 42 publications

Intratumoral Microbiota: Unraveling their Oncogenic Impact on Cancer Progression With Focus on Breast Cancer Therapeutic Outcomes

Intratumoral Microbiota: Unraveling their Oncogenic Impact on Cancer Progression With Focus on Breast Cancer Therapeutic Outcomes

Effects of Klebsiella pneumoniae Bacteriophages on IRAK3 Knockdown/Knockout THP-1 Monocyte Cell Lines

Evaluation of effectiveness of bacteriophage purification methods

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