Synthetic mRNA delivered to human cells leads to expression of Cpl-1 bacteriophage-endolysin with activity against Streptococcus pneumoniae

• Published in: Molecular therapy. Nucleic acids Vol. 35; no. 1; p. 102145
• Main Authors: Jansson, Moritz K., Nguyen, Dat Tien, Mikkat, Stefan, Warnke, Carolin, Janssen, Marc Benjamin, Warnke, Philipp, Kreikemeyer, Bernd, Patenge, Nadja
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 12-03-2024; Elsevier
• Subjects: bacterial infection; bacteriophage endolysins; Cpl-1 endolysin; infectious diseases; mRNA therapy; mRNA-based antibacterial therapy; mRNA-based endolysins; mRNA-encoded endolysins; MT: Delivery Strategies; Streptococcus pneumoniae; Cpl-1 endolysin; mRNA-encoded endolysins; infectious diseases; mRNA-based endolysins; mRNA therapy; mRNA-based antibacterial therapy; MT: Delivery Strategies; bacterial infection; Streptococcus pneumoniae; bacteriophage endolysins
• ISSN: 2162-2531; 2162-2531
• DOI: 10.1016/j.omtn.2024.102145

Endolysins are bacteriophage-encoded hydrolases that show high antibacterial activity and a narrow substrate spectrum. We hypothesize that an mRNA-based approach to endolysin therapy can overcome some challenges of conventional endolysin therapy, namely organ targeting and bioavailability. We show that synthetic mRNA applied to three human cell lines (HEK293T, A549, HepG2 cells) leads to expression and cytosolic accumulation of the Cpl-1 endolysin with activity against Streptococcus pneumoniae. Addition of a human lysozyme signal peptide sequence translocates the Cpl-1 to the endoplasmic reticulum leading to secretion (hlySP-sCpl-1). The pneumococcal killing effect of hlySP-sCpl-1 was enhanced by introduction of a point mutation to avoid N-linked-glycosylation. hlySP-sCpl-1N215D, collected from the culture supernatant of A549 cells 6 h post-transfection showed a significant killing effect and was active against nine pneumococcal strains. mRNA-based cytosolic Cpl-1 and secretory hlySP-sCpl-1N215D show potential for innovative treatment strategies against pneumococcal disease and, to our best knowledge, represent the first approach to mRNA-based endolysin therapy. We assume that many other bacterial pathogens could be targeted with this novel approach.
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RNA-based therapeutics provide innovative treatment options. Moritz K. Jansson and colleagues demonstrate proof of concept of an mRNA-based approach for bacteriophage-endolysin production in human cells. In light of the current antibiotics resistance crisis, this concept is an impulse for the urgently needed development of new antibacterial therapies.