In vitro lung delivery of bacteriophages KS4-M and ΦKZ using dry powder inhalers for treatment of Burkholderia cepacia complex and Pseudomonas aeruginosa infections in cystic fibrosis

Golshahi, Laleh; Lynch, Karlene H.; Dennis, Jonathan J.; Finlay, Warren H.

doi:10.1111/j.1365-2672.2010.04863.x

Cited by 103 publications

(85 citation statements)

References 58 publications

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“…In contrast, phage particle aerosols generated by a nebulizer and delivered via a NOID constitute a better method for alveolar phage deposition (32). Several recent studies suggested that aerosol phage therapy is not only possible but that it can be an effective method for delivering phages directly to the site of a pulmonary bacterial infection (32,33,48). In this study, immunocompromised and BCC-infected mice receiving aerosolized phage treatments exhibited significant decreases in bacterial loads within the lungs.…”

Section: Discussionmentioning

confidence: 71%

“…Although these studies have investigated phage therapy in different mouse infection models, they have done so using intranasal instillation as the delivery method (22)(23)(24)(25), which is inefficient and produces significant phage loss through ingestion. We have previously demonstrated that phages can be successfully aerosolized without damage (32,33), and utilizing this knowledge, we demonstrate here that aerosol phage therapy in mice is effective with phages delivered as a nebulized aerosol. Direct aerosolization of a therapeutic agent is the ideal method of delivery when treating a patient with a respiratory infection, and this method is an established mode of chemical drug delivery (34).…”

mentioning

confidence: 95%

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Aerosol Phage Therapy Efficacy in Burkholderia cepacia Complex Respiratory Infections

Semler

Goudie

Finlay

et al. 2014

Antimicrob Agents Chemother

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Phage therapy has been suggested as a potential treatment for highly antibiotic-resistant bacteria, such as the species of the Burkholderia cepacia complex (BCC). To address this hypothesis, experimental B. cenocepacia respiratory infections were established in mice using a nebulizer and a nose-only inhalation device. Following infection, the mice were treated with one of five B. cenocepacia-specific phages delivered as either an aerosol or intraperitoneal injection. The bacterial and phage titers within the lungs were assayed 2 days after treatment, and mice that received the aerosolized phage therapy demonstrated significant decreases in bacterial loads. Differences in phage activity were observed in vivo. Mice that received phage treatment by intraperitoneal injection did not demonstrate significantly reduced bacterial loads, although phage particles were isolated from their lung tissue. Based on these data, aerosol phage therapy appears to be an effective method for treating highly antibiotic-resistant bacterial respiratory infections, including those caused by BCC bacteria.

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Section: Discussionmentioning

confidence: 71%

mentioning

confidence: 95%

Aerosol Phage Therapy Efficacy in Burkholderia cepacia Complex Respiratory Infections

Semler

Goudie

Finlay

et al. 2014

Antimicrob Agents Chemother

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“…1) and can be assigned to the order Caudovirales and family Myoviridae based on this morphology. Phage KS4-M has been used successfully in both nebulization and lyophilization studies 22,19 and is easy to obtain in high titers.…”

Section: Bacteriophagesmentioning

confidence: 99%

“…21 Freezedrying with subsequent micronization has been successfully applied to bacteriophages for respiratory delivery. 22 An acceptable titer loss was found on processing and the in vitro performance of the resulting powder demonstrated efficient delivery by a dry powder inhaler (DPI).…”

Section: Introductionmentioning

confidence: 99%

Spray-dried Respirable Powders Containing Bacteriophages for the Treatment of Pulmonary Infections

Matinkhoo

Lynch

Dennis

et al. 2011

Journal of Pharmaceutical Sciences

113

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“…These phages are distributed widely geographically and are of interest because those infective for pathogens have been used for phage therapy (4) and/or have potential for further biocontrol applications (5)(6)(7)(8). These phages are also a curiosity, as they are remarkably complex, both at the genome and structural levels, with large virions comprised of a Tϭ27 capsid or head (9) and contractile tail that ends in an elaborate baseplate (10,11).…”

mentioning

confidence: 99%

Mutational Analysis of the Pseudomonas aeruginosa Myovirus ϕKZ Morphogenetic Protease gp175

Thomas

Black

2013

J Virol

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Pseudomonas aeruginosa myovirus KZ has a 270-kb genome within a T‫72؍‬ icosahedral capsid that contains a large, unusual, and structurally well-defined protein cylindrical inner body (IB) spanning its interior. Proteolysis forms a pivotal stage in KZ head and IB morphogenesis, with the protease gp175 cleaving at least 19 of 49 different head proteins, including the major capsid protein and five major structural IB proteins. Here we show that the purified mature form of gp175 is active and cleaves purified IB structural proteins gp93 and gp89. Expression vector synthesis and purification of the zymogen/precursor yielded an active, mature-length protease, showing independent C-terminal gp175 self-cleavage autoactivation. Mutation of either the predicted catalytic serine or histidine inactivated mature gp175, supporting its classification as a serine protease and representing the first such direct biochemical demonstration with purified protease and substrate proteins for any phage protease. These mutations also blocked self-cleavage of the precursor while allowing intermolecular gp175 processing. To confirm the cleavage specificity of gp175, we mutated three cleavage sites in gp93, which blocked proteolysis at these sites. The N-terminal propeptide of gp93 was shown to undergo more extensive proteolysis than previously identified. We found that proteolysis in gp93 progressed from the N to C terminus, while blocking cleavage sites slowed but did not eliminate downstream proteolysis. These findings were shown by informatics to be relevant to the head morphogenesis of numbers of other related IB-containing giant phages as well as to T4 and herpesviruses, which have homologous proteases.T he giant myovirus KZ, which is infective for Pseudomonas aeruginosa, is a member of the KZ-related phages that currently incorporate two genera, the KZ-like phages (KZ, PA3, and 2012-1) and the more distantly related EL-like phages (EL and OBP) (1-3). These phages are distributed widely geographically and are of interest because those infective for pathogens have been used for phage therapy (4) and/or have potential for further biocontrol applications (5-8). These phages are also a curiosity, as they are remarkably complex, both at the genome and structural levels, with large virions comprised of a Tϭ27 capsid or head (9) and contractile tail that ends in an elaborate baseplate (10, 11). The complexity of the virion is reflected in the numbers of different proteins, Ͼ60 different proteins, with nearly 50 different proteins present in the head alone, possibly the highest number for any known group of viruses (12)(13)(14). A remarkable feature of the KZ-related phages is that within the head there is a large, protein cylinder called an inner body (IB) around which the DNA is tightly spooled (15). The structure of the KZ IB was recently solved, revealing it to be approximately 24 by 105 nm and tilted at an angle of approximately 22°relative to the portal axis (16). Its structure is also regular, with an apparent 6-fold symmetry in some ...

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In vitro lung delivery of bacteriophages KS4-M and ΦKZ using dry powder inhalers for treatment of Burkholderia cepacia complex and Pseudomonas aeruginosa infections in cystic fibrosis

Abstract: Development of lactoferrin-based bacteriophage aerosol powders solidifies the ground for future research on developing novel formulations as an alternative to inhaled antibiotic therapy in patients with cystic fibrosis.

Cited by 103 publications

References 58 publications

Aerosol Phage Therapy Efficacy in Burkholderia cepacia Complex Respiratory Infections

Aerosol Phage Therapy Efficacy in Burkholderia cepacia Complex Respiratory Infections

Spray-dried Respirable Powders Containing Bacteriophages for the Treatment of Pulmonary Infections

Mutational Analysis of the Pseudomonas aeruginosa Myovirus ϕKZ Morphogenetic Protease gp175

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