Gut-Kidney Axis on Chip for Studying Effects of Antibiotics on Risk of Hemolytic Uremic Syndrome by Shiga Toxin-Producing Escherichia coli.

Yugyeong Lee,Sejoong Kim,Sungsu Park,Jong Hwan Sung,Min-Hyeok Kim,David Rodrigues Alves,Luke P Lee

doi:10.3390/toxins13110775

Abstract

Shiga toxin-producing Escherichia coli (STEC) infects humans by colonizing the large intestine, and causes kidney damage by secreting Shiga toxins (Stxs). The increased secretion of Shiga toxin 2 (Stx2) by some antibiotics, such as ciprofloxacin (CIP), increases the risk of hemolytic–uremic syndrome (HUS), which can be life-threatening. However, previous studies evaluating this relationship have been conflicting, owing to the low frequency of EHEC infection, very small number of patients, and lack of an appropriate animal model. In this study, we developed gut–kidney axis (GKA) on chip for co-culturing gut (Caco-2) and kidney (HKC-8) cells, and observed both STEC O157:H7 (O157) infection and Stx intoxication in the gut and kidney cells on the chip, respectively. Without any antibiotic treatment, O157 killed both gut and kidney cells in GKA on the chip. CIP treatment reduced O157 infection in the gut cells, but increased Stx2-induced damage in the kidney cells, whereas the gentamycin treatment reduced both O157 infection in the gut cells and Stx2-induced damage in the kidney cells. This is the first report to recapitulate a clinically relevant situation, i.e., that CIP treatment causes more damage than gentamicin treatment. These results suggest that GKA on chip is very useful for simultaneous observation of O157 infections and Stx2 poisoning in gut and kidney cells, making it suitable for studying the effects of antibiotics on the risk of HUS.

Highlights

Shiga toxin-producing Escherichia coli (STEC), including the serotype O157:H7 (O157), infects the human gastrointestinal tract, potentially leading to the development of hemolytic–uremic syndrome (HUS), a syndrome characterized by mechanical hemolytic anemia, thrombocytopenia, and kidney dysfunction [1]
Antibiotics inhibit the growth of STEC, but quinolone antibiotics, including ciprofloxacin (CIP), can increase the production of Shiga toxin 2 (Stx2) by increasing the expression of the stx2 gene
It was reported that Stx2 from O157 infection caused damage to the kidney epithelial cells, reducing their cellular integrity [31]. These results suggest that the small amounts of Stx2 produced by O157 in Caco-2 cells may be detrimental to HKC-8 cells in the chip

Summary

Introduction

Shiga toxin-producing Escherichia coli (STEC), including the serotype O157:H7 (O157), infects the human gastrointestinal tract, potentially leading to the development of hemolytic–uremic syndrome (HUS), a syndrome characterized by mechanical hemolytic anemia, thrombocytopenia, and kidney dysfunction [1]. In 2019, there was an outbreak of O157 in Brazil, and three out of 24 patients developed HUS, with one death [3]. This high mortality is partly owing to the lack of proper antibiotic treatments for patients with STEC. Antibiotics inhibit the growth of STEC, but quinolone antibiotics, including ciprofloxacin (CIP), can increase the production of Shiga toxin 2 (Stx2) by increasing the expression of the stx gene. This is a major contributor to the development of HUS [4,5]. The pentamer of the B subunit binds to the cellular receptor globotriaosylceramide-3

Methods

Results

Conclusion