Biliary atresia is the most common cause of pediatric end stage liver disease and the number one indication for pediatric liver transplantation. Because pathogenic viruses have been found in the liver of afflicted children, a proposed etiology for biliary atresia is a perinatal viral infection triggering immune mediated destruction of the biliary epithelium. The murine model of biliary atresia supports a viral pathogenesis as newborn mice infected with rhesus rotavirus (RRV) develop inflammation within the portal tract and extrahepatic bile duct obstruction. RRV targets the cholangiocyte for infection and in addition to direct cholangiocyte injury also induces Natural Killer cell mediated injury to the biliary epithelium. We have previously shown that the amino acid, arginine (R) within the sequence ?SRL? (amino acids 445-447) on the RRV VP4 protein is required for viral binding and entry into biliary epithelial cells. We developed a reverse genetics system to create a mutant of RRV (RRVVP4-R446G), which had a single amino acid change in VP4 protein compared to wild type RRV. In vitro, the mutant virus had reduced binding and infectivity in cholangiocytes. In vivo, it produced less symptoms and mortality in neonatal mice, resulting in an attenuated form of biliary atresia. We will use this mutant strain along with additional VP4 mutants to determine how RRV binds to, enters, traffics through the cell, and ultimately activates the cholangiocyte's innate immune response. We will also ascertain these VP4 mutants' ability to infect and activate plasmacytoid dendritic cells leading to Natural Killer cell activation. These complimentary approaches will generate new insight in viral induced biliary atresia.

Public Health Relevance

Biliary atresia is the most common cause of pediatric end stage liver disease and the number one indication for pediatric liver transplantation. Because pathogenic viruses have been found in the liver of afflicted children, a proposed etiology for biliary atresia is a perinatal viral infection triggering immune mediated destruction of the biliary epithelium resulting in biliary obstruction. Our goal using a unique set of rotavirus strains containing mutations in the virus attachment protein, is to determine how rotavirus activates the innate immune system resulting in bile duct obstruction, in order to develop new treatment strategies to alter the course of this challenging disease.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
2R01DK091566-06A1
Application #
9661141
Study Section
Hepatobiliary Pathophysiology Study Section (HBPP)
Program Officer
Sherker, Averell H
Project Start
2011-04-01
Project End
2022-12-31
Budget Start
2019-01-01
Budget End
2019-12-31
Support Year
6
Fiscal Year
2019
Total Cost
Indirect Cost
Name
Cincinnati Children's Hospital Medical Center
Department
Type
DUNS #
071284913
City
Cincinnati
State
OH
Country
United States
Zip Code
45229
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Mohanty, Sujit K; Donnelly, Bryan; Dupree, Phylicia et al. (2017) A Point Mutation in the Rhesus Rotavirus VP4 Protein Generated through a Rotavirus Reverse Genetics System Attenuates Biliary Atresia in the Murine Model. J Virol 91:
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Walther, Ashley; Mohanty, Sujit K; Donnelly, Bryan et al. (2015) Rhesus rotavirus VP4 sequence-specific activation of mononuclear cells is associated with cholangiopathy in murine biliary atresia. Am J Physiol Gastrointest Liver Physiol 309:G466-74
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