TRAP-seq identifies cystine/glutamate antiporter as a driver of recovery from liver injury

Understanding the molecular basis of the regenerative response following hepatic injury holds promise for improved treatment of liver diseases. Here, we report an innovative method to profile gene expression specifically in the hepatocytes that regenerate the liver following toxic injury. We used th...

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Published inThe Journal of clinical investigation Vol. 128; no. 6; pp. 2297 - 2309
Main Authors Wang, Amber W, Wangensteen, Kirk J, Wang, Yue J, Zahm, Adam M, Moss, Nicholas G, Erez, Noam, Kaestner, Klaus H
Format Journal Article
LanguageEnglish
Published United States American Society for Clinical Investigation 01.06.2018
Subjects
Acids
Analgesics
Analysis
Antioxidants
Bioinformatics
Biomedical research
Cancer
Care and treatment
Cell cycle
Cell division
Cystine
Fumarylacetoacetase
Gene expression
Genomes
Glutathione
Hepatocytes
Hepatology
Homeostasis
Liver
Liver diseases
Liver regeneration
Metabolism
Metabolites
Proteins
Purification
Recovery (Medical)
Ribonucleic acid
RNA
RNA sequencing
Rodents
Therapeutic applications
Transcription factors
Expression profiling
Molecular genetics
Mouse models
Hepatology
Gastroenterology
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Summary:Understanding the molecular basis of the regenerative response following hepatic injury holds promise for improved treatment of liver diseases. Here, we report an innovative method to profile gene expression specifically in the hepatocytes that regenerate the liver following toxic injury. We used the Fah-/- mouse, a model of hereditary tyrosinemia, which conditionally undergoes severe liver injury unless fumarylacetoacetate hydrolase (FAH) expression is reconstituted ectopically. We used translating ribosome affinity purification followed by high-throughput RNA sequencing (TRAP-seq) to isolate mRNAs specific to repopulating hepatocytes. We uncovered upstream regulators and important signaling pathways that are highly enriched in genes changed in regenerating hepatocytes. Specifically, we found that glutathione metabolism, particularly the gene Slc7a11 encoding the cystine/glutamate antiporter (xCT), is massively upregulated during liver regeneration. Furthermore, we show that Slc7a11 overexpression in hepatocytes enhances, and its suppression inhibits, repopulation following toxic injury. TRAP-seq allows cell type-specific expression profiling in repopulating hepatocytes and identified xCT, a factor that supports antioxidant responses during liver regeneration. xCT has potential as a therapeutic target for enhancing liver regeneration in response to liver injury.
Bibliography:Authorship note: AWW and KJW contributed equally to this work.
ISSN:0021-9738
1558-8238
DOI:10.1172/JCI95120