Clostridioides difficile S-Layer Protein A (SlpA) Serves as a General Phage Receptor

Royer, A.; Umansky, Andrew A; Allen, Marie-Maude; Garneau, Julian R.; Ospina-Bedoya, Maicol; Kirk, Joseph A.; Govoni, Gregory; Fagan, Robert P.; Soutourina, Olga; Fortier, Louis‐Charles

doi:10.1128/spectrum.03894-22

Cited by 5 publications

(4 citation statements)

References 53 publications

(101 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Another important limitation of current C. difficile phages is their narrow host range, implying that multiple phages will need to be combined into cocktails to cover the most clinically relevant strains of C. difficile ( 21 ). Due to recent advances in our understanding of host recognition by C. difficile phages, the surface layer protein A (SlpA) seems to be a general receptor used by many phages and diffocins ( 28 , 49 , 57 – 59 ). It is, therefore, reasonable to foresee the selection of phages based on their RBP to target C. difficile strains expressing specific SlpA isoforms.…”

Section: Discussionmentioning

confidence: 99%

The long and sinuous road to phage-based therapy of Clostridioides difficile infections

Umansky,

Fortier

2023

Front. Med.

Self Cite

View full text Add to dashboard Cite

With the antibiotic crisis and the rise in antimicrobial resistance worldwide, new therapeutic alternatives are urgently needed. Phage therapy represents one of the most promising alternatives but for some pathogens, such as Clostridioides difficile, important challenges are being faced. The perspective of phage therapy to treat C. difficile infections is complicated by the fact that no strictly lytic phages have been identified so far, and current temperate phages generally have a narrow host range. C. difficile also harbors multiple antiphage mechanisms, and the bacterial genome is often a host of one or multiple prophages that can interfere with lytic phage infection. Nevertheless, due to recent advances in phage host receptor recognition and improvements in genetic tools to manipulate phage genomes, it is now conceivable to genetically engineer C. difficile phages to make them suitable for phage therapy. Other phage-based alternatives such as phage endolysins and phage tail-like bacteriocins (avidocins) are also being investigated but these approaches also have their own limitations and challenges. Last but not least, C. difficile produces spores that are resistant to phage attacks and all current antibiotics, and this complicates therapeutic interventions. This mini-review gives a brief historical overview of phage work that has been carried out in C. difficile, presents recent advances in the field, and addresses the most important challenges that are being faced, with potential solutions.

show abstract

Section: Discussionmentioning

confidence: 99%

The long and sinuous road to phage-based therapy of Clostridioides difficile infections

Umansky,

Fortier

2023

Front. Med.

Self Cite

View full text Add to dashboard Cite

show abstract

“…Prior studies that have examined the interaction between C. difficile and phages have focussed on the host receptor [14][15][16][17][18] , leaving the reciprocal phage BRP largely uncharacterised. In phages infecting other species, the tail fibres have been shown [19][20][21][22][23] to be the RBP [19][20][21][22][23] .…”

Section: Discussionmentioning

confidence: 99%

“…To address the issue of specificity, the underlying mechanisms dictating phage binding range require investigation. Phage receptor binding protein (RBP) and host range determining factors are poorly characterised in phages infecting C. difficile, although it has been established that the counterpart host receptor is encoded by the slpA gene [14][15][16][17][18] . Part of the phage host range is its binding range, dictated by the tail components, mainly the RBP [19][20][21][22] , which is recognised as the tail fibre (or spike in other species) 23 .…”

Section: Main Introductionmentioning

confidence: 99%

A tale of two phage tails: Engineering the host range of bacteriophages infectingClostridioides difficile

Steczynska,

Kerr,

Kelly

et al. 2023

Preprint

View full text Add to dashboard Cite

Clostridioides difficile infection (CDI) is a leading cause of antibiotic-associated diarrhoea across the globe. Although treatable with a restricted number of antibiotics, the emergence of resistant variants and high relapse rates necessitate alternative countermeasures. Phage therapy represents an attractive option. However, its implementation is handicapped by the narrow host specificity of the C. difficile bacteriophages isolated to date. One strategy to rationally expand phage host range would be to make appropriate modifications to the phage receptor binding protein (RBP). Here, we identify the tail fibre as the RBP of two Myoviridae phages, ΦCD1801 and ΦCD2301, which were previously isolated and propagated using the C. difficile strains CD1801 (RT078) and CD2301 (RT014), respectively. Contrary to studies into reprogramming the host ranges of phage of other bacterial other species, exchanging the tail fibre genes (tcf/tfp) alone between the two phage was insufficient to change host specificity. Rather, alterations to host range were dependent their exchange together with a putative chaperone encoded by hyp, localised adjacent to the tail fibre gene. Capitalising on this discovery, CRISPR/Cas9 was used to change the host range of one phage to that of the other by swapping the respective tcf/tfp and hyp genes. Significantly, one of the resulting mutants, surpassed both parental phages in terms of host range and efficiency of infection. This is the first time that genome engineering has successfully expanded the host range of a C. difficile phage, a prerequisite for implementing phage for the treatment of CDI.

show abstract

“…In vivo, it is proposed to play a role in molecular sieving [18] and ion trapping, protecting the organism from antimicrobial peptides and bacteriolytic enzymes produced in response to infection [19,20]. The S-layer has also been shown to be a key target in bacteriophage predation [20][21][22].…”

Section: Introductionmentioning

confidence: 99%

An intact S-layer is advantageous to Clostridioides difficile within the host

et al. 2023

Self Cite

View full text Add to dashboard Cite

Clostridioides difficile is responsible for substantial morbidity and mortality in antibiotically-treated, hospitalised, elderly patients, in which toxin production correlates with diarrhoeal disease. While the function of these toxins has been studied in detail, the contribution of other factors, including the paracrystalline surface layer (S-layer), to disease is less well understood. Here, we highlight the essentiality of the S-layer in vivo by reporting the recovery of S-layer variants, following infection with the S-layer-null strain, FM2.5. These variants carry either correction of the original point mutation, or sequence modifications which restored the reading frame, and translation of slpA. Selection of these variant clones was rapid in vivo, and independent of toxin production, with up to 90% of the recovered C. difficile population encoding modified slpA sequence within 24 h post infection. Two variants, subsequently named FM2.5varA and FM2.5varB, were selected for study in greater detail. Structural determination of SlpA from FM2.5varB indicated an alteration in the orientation of protein domains, resulting in a reorganisation of the lattice assembly, and changes in interacting interfaces, which might alter function. Interestingly, variant FM2.5varB displayed an attenuated, FM2.5-like phenotype in vivo compared to FM2.5varA, which caused disease severity more comparable to that of R20291. Comparative RNA sequencing (RNA-Seq) analysis of in vitro grown isolates revealed large changes in gene expression between R20291 and FM2.5. Downregulation of tcdA/tcdB and several genes associated with sporulation and cell wall integrity may account for the reported attenuated phenotype of FM2.5 in vivo. RNA-seq data correlated well with disease severity with the more virulent variant, FM2.5varA, showing s similar profile of gene expression to R20291 in vitro, while the attenuated FM2.5varB showed downregulation of many of the same virulence associated traits as FM2.5. Cumulatively, these data add to a growing body of evidence that the S-layer contributes to C. difficile pathogenesis and disease severity.

show abstract

Clostridioides difficile S-Layer Protein A (SlpA) Serves as a General Phage Receptor

Cited by 5 publications

References 53 publications

The long and sinuous road to phage-based therapy of Clostridioides difficile infections

The long and sinuous road to phage-based therapy of Clostridioides difficile infections

A tale of two phage tails: Engineering the host range of bacteriophages infectingClostridioides difficile

An intact S-layer is advantageous to Clostridioides difficile within the host

Contact Info

Product

Resources

About