Improved Biodistribution and Extended Serum Half-Life of a Bacteriophage Endolysin by Albumin Binding Domain Fusion

Seijsing, Johan; Sobieraj, Anna M.; Keller, Nadia; Shen, Yang; Loessner, Martin J.; Schmelcher, Mathias

doi:10.3389/fmicb.2018.02927

Cited by 42 publications

(48 citation statements)

References 62 publications

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“…In most cases, modification with an ABD significantly reduced the activity off the enzymes in TRAs. Parental PGH activities were increased by adding HSA, as previously observed for other staphylolytic PGHs ( 41 , 58 ) ( Fig. 2A and B ).…”

Section: Resultssupporting

confidence: 85%

“…For example, the half-life of lysostaphin was previously found to be 1 h in mice ( 36 ) and 1.5 h in rats ( 37 ), that of Cpl-1 was found to be 20.5 min in mice ( 38 ), and that of SAL200 was found to be 0.3 to 9.7 h in monkeys ( 39 ) and 2.4 to 22.8 min in humans ( 40 ). The longest half-life of 23 h was reported for an engineered version of LysK, CSL[Eu], in mice ( 41 ). Elimination of proteins from the organism occurs through a combination of processes, which include general proteolysis, endocytosis and lysosomal degradation, renal filtration, and hepatic elimination ( 42 ).…”

Section: Introductionmentioning

confidence: 99%

“…Proteins covalently fused to ABD can form a high-affinity complex with HSA, which is then rescued from renal filtration due to its large hydrodynamic volume and is protected from lysosomal degradation by FcRn recycling ( 49 ). For LysK ( 41 ) and lysostaphin ( 37 ), this resulted in significant extension of their half-lives ( 37 , 41 ) and in other improvements in pharmacodynamics ( 37 ). While these results are encouraging, no studies have shown that half-life extension of PGHs translates into enhanced efficacy in infection models.…”

Section: Introductionmentioning

confidence: 99%

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Engineering of Long-Circulating Peptidoglycan Hydrolases Enables Efficient Treatment of Systemic Staphylococcus aureus Infection

Sobieraj

Huemer

Zinsli

et al. 2020

mBio

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Staphylococcus aureus is a human pathogen causing life-threatening diseases. The increasing prevalence of multidrug-resistant S. aureus infections is a global health concern, requiring development of novel therapeutic options. Peptidoglycan-degrading enzymes (peptidoglycan hydrolases, PGHs) have emerged as a highly effective class of antimicrobial proteins against S. aureus and other pathogens. When applied to Gram-positive bacteria, PGHs hydrolyze bonds within the peptidoglycan layer, leading to rapid bacterial death by lysis. This activity is highly specific and independent of the metabolic activity of the cell or its antibiotic resistance patterns. However, systemic application of PGHs is limited by their often low activity in vivo and by an insufficient serum circulation half-life. To address this problem, we aimed to extend the half-life of PGHs selected for high activity against S. aureus in human serum. Half-life extension and increased serum circulation were achieved through fusion of PGHs to an albumin-binding domain (ABD), resulting in high-affinity recruitment of human serum albumin and formation of large protein complexes. Importantly, the ABD-fused PGHs maintained high killing activity against multiple drug-resistant S. aureus strains, as determined by ex vivo testing in human blood. The top candidate, termed ABD_M23, was tested in vivo to treat S. aureus-induced murine bacteremia. Our findings demonstrate a significantly higher efficacy of ABD_M23 than of the parental M23 enzyme. We conclude that fusion with ABD represents a powerful approach for half-life extension of PGHs, expanding the therapeutic potential of these enzybiotics for treatment of multidrug-resistant bacterial infections. IMPORTANCE Life-threatening infections with Staphylococcus aureus are often difficult to treat due to the increasing prevalence of antibiotic-resistant bacteria and their ability to persist in protected niches in the body. Bacteriolytic enzymes are promising new antimicrobials because they rapidly kill bacteria, including drug-resistant and persisting cells, by destroying their cell wall. However, when injected into the bloodstream, these enzymes are not retained long enough to clear an infection. Here, we describe a modification to increase blood circulation time of the enzymes and enhance treatment efficacy against S. aureus-induced bloodstream infections. This was achieved by preselecting enzyme candidates for high activity in human blood and coupling them to serum albumin, thereby preventing their elimination by kidney filtration and blood vessel cells.

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Section: Resultssupporting

confidence: 85%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Engineering of Long-Circulating Peptidoglycan Hydrolases Enables Efficient Treatment of Systemic Staphylococcus aureus Infection

Sobieraj

Huemer

Zinsli

et al. 2020

mBio

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“…Our approach and similar screening approaches may serve to find new lead compounds against intracellular MRSA but might also be transferred to PGHs of other pathogens, such as Streptococcus pyogenes, Listeria monocytogenes, and Bacillus anthracis, which can occur intracellularly (12). Furthermore, such strategies may be used to identify suitable PGHs for applications in various different niches, for example, treatment of bloodstream infections (41).…”

Section: Discussionmentioning

confidence: 99%

Targeting Hidden Pathogens: Cell-Penetrating Enzybiotics Eradicate Intracellular Drug-Resistant Staphylococcus aureus

Röhrig

Huemer

Lorgé

et al. 2020

mBio

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Staphylococcus aureus is a major concern in human health care, mostly due to the increasing prevalence of antibiotic resistance. Intracellular localization of S. aureus plays a key role in recurrent infections by protecting the pathogens from antibiotics and immune responses. Peptidoglycan hydrolases (PGHs) are highly specific bactericidal enzymes active against both drug-sensitive and -resistant bacteria. However, PGHs able to effectively target intracellular S. aureus are not yet available. To overcome this limitation, we first screened 322 recombineered PGHs for staphylolytic activity under conditions found inside eukaryotic intracellular compartments. The most active constructs were modified by fusion to different cell-penetrating peptides (CPPs), resulting in increased uptake and enhanced intracellular killing (reduction by up to 4.5 log units) of various S. aureus strains (including methicillin-resistant S. aureus [MRSA]) in different tissue culture infection models. The combined application of synergistic PGH-CPP constructs further enhanced their intracellular efficacy. Finally, synergistically active PGH-CPP cocktails reduced the total S. aureus by more than 2.2 log units in a murine abscess model after peripheral injection. Significantly more intracellular bacteria were killed by the PGH-CPPs than by the PGHs alone. Collectively, our findings show that CPP-fused PGHs are effective novel protein therapeutics against both intracellular and drug-resistant S. aureus. IMPORTANCE The increasing prevalence of antibiotic-resistant bacteria is one of the most urgent problems of our time. Staphylococcus aureus is an important human pathogen that has acquired several mechanisms to evade antibiotic treatment. In addition, S. aureus is able to invade and persist within human cells, hiding from the immune response and antibiotic therapies. For these reasons, novel antibacterial strategies against these pathogens are needed. Here, we developed lytic enzymes which are able to effectively target drug-resistant and intracellular S. aureus. Fusion of these so-called enzybiotics to cell-penetrating peptides enhanced their uptake and intracellular bactericidal activity in cell culture and in an abscess mouse model. Our results suggest that cell-penetrating enzybiotics are a promising new class of therapeutics against staphylococcal infections.

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“…At least one report indicates that this problem might be more relevant in genetically manipulated phages. 87) A similar phenomenon is seen with endolysins; however, a recent report indicates that the engineering of an endolysin in order to add an albumin-binding domain extended the half-life of the endolysin. 88) Another area that could become relevant, if phage therapy advances toward widespread use, is the study of storage conditions and administration routes.…”

Section: Future Challenges and Projectionsmentioning

confidence: 68%

Engineered Bacteriophages for Practical Applications

Pizarro-Bäuerle

Ando

2020

Biological & Pharmaceutical Bulletin

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The diversity of advanced genetic engineering techniques that have become available in recent years has enabled a more precise manipulation of genes and genomes. Among these, bacteriophage genomes stand out as an interesting target due to their dependence on a host for replication, which previously complicated their manipulation, and due as well to the many possible fields in which they can be used. In this review, we highlight recent applications for which genetically modified bacteriophages are being employed: as phage therapy in medicine, animal industries and agricultural settings; as a source of new antimicrobials; as biosensors for research, health and environmental purposes; and as genetic engineering tools themselves.

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Improved Biodistribution and Extended Serum Half-Life of a Bacteriophage Endolysin by Albumin Binding Domain Fusion

Cited by 42 publications

References 62 publications

Engineering of Long-Circulating Peptidoglycan Hydrolases Enables Efficient Treatment of Systemic Staphylococcus aureus Infection

Engineering of Long-Circulating Peptidoglycan Hydrolases Enables Efficient Treatment of Systemic Staphylococcus aureus Infection

Targeting Hidden Pathogens: Cell-Penetrating Enzybiotics Eradicate Intracellular Drug-Resistant Staphylococcus aureus

Engineered Bacteriophages for Practical Applications

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