Freeze-dried fecal samples are biologically active after long-lasting storage and suited to fecal microbiota transplantation in a preclinical murine model of Clostridioides difficile infection
Julie Reygner, Christine Charrueau, Johanne Delannoy, Camille Mayeur, Véronique Robert, Céline Cuinat, Thierry Meylheuc, Aurélie Mauras, Jérémy Augustin, Ioannis Nicolis, Morgane Modoux, Francisca Joly, Anne-Judith Waligora-Dupriet, Muriel Thomas, Nathalie Kapel, Julie Reygner, Christine Charrueau, Johanne Delannoy, Camille Mayeur, Véronique Robert, Céline Cuinat, Thierry Meylheuc, Aurélie Mauras, Jérémy Augustin, Ioannis Nicolis, Morgane Modoux, Francisca Joly, Anne-Judith Waligora-Dupriet, Muriel Thomas, Nathalie Kapel
Abstract
Fecal microbiota transplantation is now recommended for treating recurrent forms of Clostridioides difficile infection. Recent studies have reported protocols using capsules of either frozen or freeze-dried stool allowing oral administration in in- and out-patient settings. However, a central question remains the viability, engraftment, and efficacy of the microbiome over time during storage life. This study shows that both the freeze-drying and freezing procedures for fecal samples allowed preserving viability, short-chain fatty acids concentration, and anti-Clostridioides difficile properties of microbiota without significant alteration after storage for 12 months. Fecal transplantation with freeze-dried microbiota allowed engraftment of microbiota leading to clearance of Clostridioides difficile infection in a preclinical murine model with a survival rate of 70% versus 53-60% in mice treated with frozen inocula, and 20% in the untreated group. Moreover, the freeze-dried powder can be used to fill oral hard capsules using a very low amount (0.5%) of glidant excipient, allowing oral formulation. Altogether, this study showed that freeze-dried inocula can be used for the treatment of Clostridioides difficile infection with long-lasting stability of the fecal microbiota. This formulation facilitates biobanking and allows the use of hard capsules, an essential step to simplify patient access to treatment.
Keywords: Clostridioides difficile; Fecal microbiota transplant; freeze-dried microbiota; frozen microbiota; pre-clinical model.
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References
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