Enhanced Wound Healing by Recombinant Escherichia coli Nissle 1917 via Human Epidermal Growth Factor Receptor in Human Intestinal Epithelial Cells: Therapeutic Implication Using Recombinant Probiotics

Abstract

The gastrointestinal mucosa has a remarkable ability to repair damage with the support of epidermal growth factor (EGF), which stimulates epithelial migration and proliferative reepithelialization. For the treatment of mucosal injuries, it is important to develop efficient methods for the localized delivery of mucoactive biotherapeutics. The basic idea in the present study came from the assumption that an intestinal probiotic vehicle can carry and deliver key recombinant medicinal proteins to the injured epithelial target in patients with intestinal ulcerative diseases, including inflammatory bowel disease. The study was focused on the use of the safe probiotic E. coli Nissle 1917, which was constructed to secrete human EGF in conjunction with the lipase ABC transporter recognition domain (LARD). Using the in vitro physically wounded monolayer model, ABC transporter-mediated EGF secretion by probiotic E. coli Nissle 1917 was demonstrated to enhance the wound-healing migration of human enterocytes. Moreover, the epithelial wound closure was dependent on EGF receptor-linked activation, which exclusively involved the subsequent signaling pathway of the mitogen-activated protein kinase kinase (MEK) extracellular-related kinases 1 and 2 (ERK1/2). In particular, the migrating frontier of the wounded edge displayed the strongest EGF receptor-linked signaling activation in the presence of the recombinant probiotic. The present study provides a basis for the clinical application of human recombinant biotherapeutics via an efficient, safe probiotic vehicle.

Probiotics: Properties, Examples, and Specific Applications

“…Although engineering Gram-positive probiotic strains like LAB to deliver molecules of interest has been a relatively uncomplicated affair because of the comparatively simple nature of their cell walls, Gram-negative probiotics like EcN have also been engineered to secrete molecules (Rao et al 2005;Choi et al 2012). In an attempt to block infection with the human immunodeficiency virus (HIV), EcN was engineered to secrete a hybrid peptide comprising the HIV protein gp41 (which catalyzes receptor-mediated membrane fusion) and the EcN hemolysin A (Hly), which allows direct export from the EcN cytoplasm into the extracellular medium.…”

“…In this scenario, the persistent colonization of EcN constitutes an advantage over LAB, which only transiently colonize the gut mucosa. In another recent study, EcN was engineered to deliver epidermal growth factor as a means to enhance wound healing (Choi et al 2012). Therefore, although LAB have been the probiotics of choice for targeted delivery of molecules, EcN may provide distinct advantages when persistent colonization is desirable.…”

Effects of epidermal growth factor-loaded mucoadhesive films on wounded oral tissue rafts

“…It also promotes formation of granulation tissue and stimulates fibroblast motility (11). In vitro studies show that EGF plays a significant role in healing of mucosal tissues by promoting migration and proliferation of epithelial cells in the oral cavity and intestinal mucosa (12, 13). Exogenous EGF improved tongue wound healing in sialodenectomized mice, i.e.…”

The use of probiotics and safety concerns: A review

“…A proposed scheme for safety assessment of probiotics is presented in Table 1. Most stringent studies have to be completed for genetically modified strains intended for human consumption before commercialization (Choi et al, 2012;Sorokulova, 2008;Salminen et al, 1996). Table 1.…”