Activation of Wnt signaling by amniotic fluid stem cell-derived extracellular vesicles attenuates intestinal injury in experimental necrotizing enterocolitis

Bo Li,Carol Lee,Paul Delgado Olguin,Mashriq Alganabi,Kathene C Johnson‐Henry ,Richard Wu ,Paolo De Coppi,Hideaki Miyake ,Francesca Nascimben,Michael F Maalouf ,Joshua S O'connell ,Marissa Cadete,Augusto Zani,Steven R Botts,Elke Zani‐Ruttenstock ,Pekka Määttänen ,Alison Hock,Niloofar Ganji,Yong Chen,Yuhki Koike,Lina Antounians,Philip M Sherman,Adam Minich,Simon Eaton,Agostino Pierro

doi:10.1038/s41419-020-02964-2

Abstract

Necrotizing enterocolitis (NEC) is a devastating intestinal disease primarily affecting preterm neonates and causing high morbidity, high mortality, and huge costs for the family and society. The treatment and the outcome of the disease have not changed in recent decades. Emerging evidence has shown that stimulating the Wnt/β-catenin pathway and enhancing intestinal regeneration are beneficial in experimental NEC, and that they could potentially be used as a novel treatment. Amniotic fluid stem cells (AFSC) and AFSC-derived extracellular vesicles (EV) can be used to improve intestinal injury in experimental NEC. However, the mechanisms by which they affect the Wnt/β-catenin pathway and intestinal regeneration are unknown. In our current study, we demonstrated that AFSC and EV attenuate NEC intestinal injury by activating the Wnt signaling pathway. AFSC and EV stimulate intestinal recovery from NEC by increasing cellular proliferation, reducing inflammation and ultimately regenerating a normal intestinal epithelium. EV administration has a rescuing effect on intestinal injury when given during NEC induction; however, it failed to prevent injury when given prior to NEC induction. AFSC-derived EV administration is thus a potential emergent novel treatment strategy for NEC.

Highlights

Necrotizing enterocolitis (NEC) is one of the most devastating diseases in newborn infants, primarily affecting preterm and low birth weight neonates[1,2]
We recently demonstrated that Intestinal stem cells (ISC) impairment in NEC is caused by defective Wnt signaling[7], a pathway that plays a central role in the regulation of stem cells and tissue homeostasis[8,9], including gastrointestinal epithelium[10]
NEC induction was conducted from postnatal (p) day 5 to 9 with Amniotic fluid stem cells (AFSC) intraperitoneal injection given on p6 and p7, and animals sacrificed on p9 (Fig. 1a)

Summary

Introduction

Necrotizing enterocolitis (NEC) is one of the most devastating diseases in newborn infants, primarily affecting preterm and low birth weight neonates[1,2]. ISC depletion correlates with severe gut damage during NEC. Administration of amniotic fluid to preterm born neonatal piglets increases body weight and prompts postnatal development[14,15]. Amniotic fluid attenuates the severity of experimental NEC by reducing gut inflammation[16] and inhibiting Tolllike receptor (TLR)-4 mediated signaling[17]. These beneficial effects may be due to the presence of amniotic fluid stem cells (AFSC). In neonatal rats with experimental NEC, we reported that AFSC administration decreased apoptosis and mucosal inflammation, and increased cell proliferation and migration through a paracrine mechanism[18]. The mechanism(s) by which AFSC elicited their protective effects during intestinal regeneration during NEC remains unclear

Methods

Results

Conclusion