Engineered Lactobacillus paracasei Producing Palmitoylethanolamide (PEA) Prevents Colitis in Mice.

Giuseppe Esposito,Chiara Corpetti,Giovanni Esposito,Alessandro Del Re,Marcella Pesce,Jie Lu,Luisa Seguella,Giovanni Sarnelli,Fatima Domenica Elisa De Palma,Walter Sanseverino

doi:10.3390/ijms22062945

Abstract

Palmitoylethanolamide (PEA) is an N-acylethanolamide produced on-demand by the enzyme N-acylphosphatidylethanolamine-preferring phospholipase D (NAPE-PLD). Being a key member of the larger family of bioactive autacoid local injury antagonist amides (ALIAmides), PEA significantly improves the clinical and histopathological stigmata in models of ulcerative colitis (UC). Despite its safety profile, high PEA doses are required in vivo to exert its therapeutic activity; therefore, PEA has been tested only in animals or human biopsy samples, to date. To overcome these limitations, we developed an NAPE-PLD-expressing Lactobacillus paracasei F19 (pNAPE-LP), able to produce PEA under the boost of ultra-low palmitate supply, and investigated its therapeutic potential in a murine model of UC. The coadministration of pNAPE-LP and palmitate led to a time-dependent release of PEA, resulting in a significant amelioration of the clinical and histological damage score, with a significantly reduced neutrophil infiltration, lower expression and release of pro-inflammatory cytokines and oxidative stress markers, and a markedly improved epithelial barrier integrity. We concluded that pNAPE-LP with ultra-low palmitate supply stands as a new method to increase the in situ intestinal delivery of PEA and as a new therapeutic able of controlling intestinal inflammation in inflammatory bowel disease.

Highlights

In palmitate; native Lactobacillus paracasei (pLP), no detectable levels of released PEA were observed at the same time points, even when the medium was enriched with 0.0003 μg/mL of palmitate (Figure 1A)
Considered as an innocent bystander for decades, accumulating evidence has clearly demonstrated its pivotal role in regulating several aspects of intestinal homeostasis, including mucosal integrity and inflammation [19,20]
On the basis of such experimental paradigm, here, we demonstrated the feasibility of integrating a genetically-engineered probiotic, able to biosynthesize human NAPE-PLD, into the murine microbiota, and evaluated its effects on colonic inflammation in a wellvalidated mouse model of acute colitis, using Lactobacillus paracasei F19 spp., a widely used probiotic in clinical settings, that is featured by its peculiar genetic stability

Summary

Introduction

Palmitoylethanolamide (PEA) is a naturally-produced lipid derived from the hydrolysis of its phospholipid precursor, by N-acylphosphatidylethanolamine-specific phospholipase D (NAPE-PLD) [1,2,3]. PEA exerts potent anti-inflammatory effects, and it has been shown to improve intestinal inflammation, following both intraperitoneal and oral administration [5], in animal models of colitis. Biopsies from patients with ulcerative colitis (UC) [6,7,8], with the peroxisome proliferator activated receptor α (PPARα) being one of the key receptors mediating these effects [9]. In IBD, an altered PEA turnover with relative down-expression of NAPE-PLD and overexpression of its degrading enzymes led to the postulation of an impairment of the acylethanolamide–PPARα anti-inflammatory axis in patients with active UC [11]

Methods

Results

Conclusion