Sodium-bile acid co-transporter is crucial for survival of a carcinogenic liver fluke Clonorchis sinensis in the bile

Author summary Clonorchiasis is a neglected tropical disease caused by infection with the liver fluke Clonorchis sinensis. C. sinensis is a biological carcinogen causing cholangiocarcinoma in humans. Juvenile worms inhabit and grow to adults in the bile ducts. Bile acids in the bile are double-edged molecules; they promote metabolism, but differences in their concentration across the cell membrane could lead to bile intoxication. The sodium-bile acid co-transporter of C. sinensis (CsSBAT) is indispensable for maintaining its normal physiology and bile detoxification in the bile duct. However, information related to the molecular and biological characteristics of the SBAT of liver flukes is not available. Here, we cloned CsSBAT for the first time in trematodes and characterized its tertiary structure and physiological functions. The sequential and structural properties of CsSBAT were similar to the apical sodium-bile acid co-transporter found in mammalian intestines. CsSBAT shared a mesenchymal tissue distribution with Na+-taurocholate co-transporting polypeptide in the hepatocytes adjacent to the bile ducts. Bile acids accumulated in C. sinensis adults when CsSBAT was inhibited, causing their death. This information might promote further studies on the physiological functions of SBAT and other trematode bile transporters and open new avenues toward developing novel anthelminthic drugs. The liver fluke Clonorchis sinensis inhabits the bile ducts, where bile concentration disparities across the fluke cell membrane can cause bile intoxication. Sodium-bile acid co-transporter (SBAT) plays a crucial role in bile acid recycling. The process by which SBAT imports bile acids is electrically coupled to sodium ion co-transportation. Here, we report that the SBAT of C. sinensis (CsSBAT) is involved in bile acid transportation. CsSBAT cDNA encoded a putative polypeptide of 546 amino acid residues. Furthermore, CsSBAT consisted of ten putative transmembrane domains, and its 3D structure was predicted to form panel and core domains. The CsSBAT had one bile acid- and three Na+-binding sites, enabling coordination of a symport process. CsSBAT was mainly localized in the mesenchymal tissue throughout the fluke body and sparsely localized in the basement of the tegument, intestinal epithelium, and excretory bladder wall. Bile acid permeated into the adult flukes in a short time and remained at a low concentration level. Bile acid accumulated inside the mesenchymal tissue when CsSBAT was inhibited using polyacrylic acid-tetradeoxycholic acid conjugate. The accumulated bile acid deteriorated the C. sinensis adults leading to death. CsSBAT silencing shortened the lifespan of the fluke when it was placed into bile. Taken together, we propose that CsSBAT transports bile acids in the mesenchymal tissue and coordinate with outward transporters to maintain bile acid homeostasis of C. sinensis adults, contributing to C. sinensis survival in the bile environment.