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Evaluation of the HBV liver reservoir with fine needle aspirates
Barbara Testoni1,2,3, Armando Andres Roca Suarez1,2,3, Arianna Battisti4
1INSERM U1052, CNRS UMR-5286, Cancer Research Center of Lyon (CRCL), Lyon, France.
Background & Aims:
Finite duration of treatment associated with HBsAg loss is the current goal for improved therapeutic approaches against chronic HBV infection, as it indicates elimination or durable inactivation of intrahepatic covalently closed circular DNA (cccDNA). To assist drug development, the definition of early predictive markers of HBsAg loss by assessing their value in reflecting intrahepatic cccDNA levels and transcriptional activity is essential. Fine needle aspirates (FNAs) have recently emerged as a less invasive alternative to core liver biopsy (CLB) and showed to be useful for investigating intrahepatic immune responses. The aim of this study was to optimise and validate the use of FNA vs. CLB to evaluate the intrahepatic viral reservoir.
Methods:
Paired FNA/CLB samples were obtained from patients with HBeAg+ chronic hepatitis (n = 4), HBeAg- chronic hepatitis (n = 4), and HBeAg- chronic infection (n = 1). One HBeAg+ patient was undergoing tenofovir treatment. HBV 3.5-kb RNA and cccDNA were quantified by droplet digital PCR.
Results:
cccDNA was quantifiable in all but one FNA/CLB pair, showing the highest levels in untreated HBeAg+ patients, except for the tenofovir-treated patient. Similarly, 3.5-kb RNA was detectable in all but one FNA sample and showed higher levels in HBeAg+ patients. When comparing cccDNA and 3.5-kb RNA quantification in FNA vs. CLB samples, no statistically significant differences were identified.
Conclusions:
These results demonstrate the possibility to quantify cccDNA and assess its transcriptional activity in patients with chronic hepatitis B by combining FNA and droplet digital PCR. This supports the use of FNA in clinical trials to evaluate the intrahepatic viral reservoir during the development of new antivirals and immunomodulatory agents.
Impact And Implications:
Chronic hepatitis B infection is characterised by a complex interplay between immune responses and viral replication in the liver, which determines the long-term outcome of the disease. In this study, we show that fine needle aspiration of the liver, a less-invasive alternative to core biopsies, allows the assessment of the hepatic viral reservoir.
Insights
Fine needle aspiration (FNA) can effectively measure the hepatitis B virus (HBV) reservoir in the liver. This less invasive method aids in developing new HBV treatments by assessing intrahepatic covalently closed circular DNA (cccDNA).
Area of Science:
- Hepatology
- Virology
- Molecular Biology
Background:
- Chronic hepatitis B (CHB) treatment aims for HBsAg loss, indicating elimination of intrahepatic covalently closed circular DNA (cccDNA).
- Early predictive markers reflecting cccDNA levels and activity are crucial for drug development.
- Fine needle aspiration (FNA) offers a less invasive alternative to core liver biopsy (CLB) for assessing intrahepatic immune responses.
Purpose of the Study:
- To optimize and validate the use of FNA versus CLB for evaluating the intrahepatic viral reservoir in CHB patients.
- To assess the utility of FNA in quantifying cccDNA and its transcriptional activity.
Main Methods:
- Paired FNA and CLB samples were collected from CHB patients across different stages (HBeAg+, HBeAg-, chronic infection).
- Hepatitis B virus (HBV) 3.5-kb RNA and cccDNA were quantified using droplet digital PCR (ddPCR).
- One patient was undergoing tenofovir treatment for comparison.
Main Results:
- cccDNA was quantifiable in most FNA/CLB pairs, with highest levels in untreated HBeAg+ patients.
- HBV 3.5-kb RNA was detectable in most FNA samples, showing higher levels in HBeAg+ patients.
- No significant differences were found in cccDNA and 3.5-kb RNA quantification between FNA and CLB samples.
Conclusions:
- FNA combined with ddPCR enables quantification of cccDNA and assessment of its transcriptional activity in CHB patients.
- FNA is a viable tool for clinical trials evaluating the intrahepatic viral reservoir during the development of new HBV antivirals and immunomodulatory agents.
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