iPSC-hepatocyte organoids as a novel platform to predict AAV gene therapy efficacy

Adeno-associated virus (AAV) vectors are widely used in gene therapy, particularly for liver-targeted treatments. However, predicting human-specific outcomes, such as transduction efficiency and hepatotoxicity, remains challenging. Reliable in vitro models are urgently needed to bridge the gap betwe...

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Main Authors: Estelle Berreur, Giacomo Lazzaroni, Cyrill Roth, Marco Zihlmann, Martina Stirn, Ramona Matheis, Rebecca Xicluna, Ekaterina Breous-Nystrom, Adrian B. Roth
Format: Article
Language:English
Published: Elsevier 2025-06-01
Series:Molecular Therapy: Methods & Clinical Development
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Online Access:http://www.sciencedirect.com/science/article/pii/S2329050125000622
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author Estelle Berreur
Giacomo Lazzaroni
Cyrill Roth
Marco Zihlmann
Martina Stirn
Ramona Matheis
Rebecca Xicluna
Ekaterina Breous-Nystrom
Adrian B. Roth
author_facet Estelle Berreur
Giacomo Lazzaroni
Cyrill Roth
Marco Zihlmann
Martina Stirn
Ramona Matheis
Rebecca Xicluna
Ekaterina Breous-Nystrom
Adrian B. Roth
author_sort Estelle Berreur
collection DOAJ
description Adeno-associated virus (AAV) vectors are widely used in gene therapy, particularly for liver-targeted treatments. However, predicting human-specific outcomes, such as transduction efficiency and hepatotoxicity, remains challenging. Reliable in vitro models are urgently needed to bridge the gap between preclinical studies and clinical applications. This study presents the first comparative evaluation of AAV transduction across multiple induced pluripotent stem cell (iPSC)-derived hepatocyte organoid donors, offering a novel platform for assessing vector performance in human liver models. The transduction efficiency and hepatotoxicity of eight AAV serotypes (AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV8, and AAV9) were tested in iPSC-derived liver organoids and hepatic cell lines (HepG2 and HepaRG). AAV6 and AAV8 exhibited the highest transduction efficiency in organoids, while AAV4 and AAV5 were the least effective. Transduction variability was observed across different donors and cell lines. Notably, no significant hepatotoxicity, measured by AST (aspartate aminotransferase) release and viability measurements, was observed, indicating that AAVs do not induce immediate liver damage in vitro. This study introduces iPSC-derived hepatocyte organoids as a novel and effective tool for predicting AAV transduction efficiency and safety, with potential to enhance the translation of gene therapies to clinical applications.
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spelling doaj-art-e2e91e27b4024ef88e8482e04dbaa44e2025-08-20T03:14:35ZengElsevierMolecular Therapy: Methods & Clinical Development2329-05012025-06-0133210146710.1016/j.omtm.2025.101467iPSC-hepatocyte organoids as a novel platform to predict AAV gene therapy efficacyEstelle Berreur0Giacomo Lazzaroni1Cyrill Roth2Marco Zihlmann3Martina Stirn4Ramona Matheis5Rebecca Xicluna6Ekaterina Breous-Nystrom7Adrian B. Roth8Roche Pharma Research and Early Development, Roche Innovation Center Basel, CH-4070 Basel, Switzerland; Corresponding author: Estelle Berreur, Roche Pharma Research and Early Development, Roche Innovation Center Basel, Grenzacherstrasse 124, CH-4070 Basel, Switzerland.Roche Pharma Research and Early Development, Roche Innovation Center Basel, CH-4070 Basel, SwitzerlandInstitute of Human Biology, Roche Innovation Center Basel, CH-4070 Basel, SwitzerlandRoche Pharma Research and Early Development, Roche Innovation Center Basel, CH-4070 Basel, SwitzerlandRoche Pharma Research and Early Development, Roche Innovation Center Basel, CH-4070 Basel, SwitzerlandRoche Pharma Research and Early Development, Roche Innovation Center Basel, CH-4070 Basel, SwitzerlandRoche Pharma Research and Early Development, Roche Innovation Center Basel, CH-4070 Basel, SwitzerlandRoche Pharma Research and Early Development, Roche Innovation Center Basel, CH-4070 Basel, SwitzerlandPrecision Safety, Pharma Product Development, Roche Innovation Center Basel, CH-4070 Basel, SwitzerlandAdeno-associated virus (AAV) vectors are widely used in gene therapy, particularly for liver-targeted treatments. However, predicting human-specific outcomes, such as transduction efficiency and hepatotoxicity, remains challenging. Reliable in vitro models are urgently needed to bridge the gap between preclinical studies and clinical applications. This study presents the first comparative evaluation of AAV transduction across multiple induced pluripotent stem cell (iPSC)-derived hepatocyte organoid donors, offering a novel platform for assessing vector performance in human liver models. The transduction efficiency and hepatotoxicity of eight AAV serotypes (AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV8, and AAV9) were tested in iPSC-derived liver organoids and hepatic cell lines (HepG2 and HepaRG). AAV6 and AAV8 exhibited the highest transduction efficiency in organoids, while AAV4 and AAV5 were the least effective. Transduction variability was observed across different donors and cell lines. Notably, no significant hepatotoxicity, measured by AST (aspartate aminotransferase) release and viability measurements, was observed, indicating that AAVs do not induce immediate liver damage in vitro. This study introduces iPSC-derived hepatocyte organoids as a novel and effective tool for predicting AAV transduction efficiency and safety, with potential to enhance the translation of gene therapies to clinical applications.http://www.sciencedirect.com/science/article/pii/S2329050125000622AAV vectorsliveriPSCorganoidsAAV transductionAAV serotype
spellingShingle Estelle Berreur
Giacomo Lazzaroni
Cyrill Roth
Marco Zihlmann
Martina Stirn
Ramona Matheis
Rebecca Xicluna
Ekaterina Breous-Nystrom
Adrian B. Roth
iPSC-hepatocyte organoids as a novel platform to predict AAV gene therapy efficacy
Molecular Therapy: Methods & Clinical Development
AAV vectors
liver
iPSC
organoids
AAV transduction
AAV serotype
title iPSC-hepatocyte organoids as a novel platform to predict AAV gene therapy efficacy
title_full iPSC-hepatocyte organoids as a novel platform to predict AAV gene therapy efficacy
title_fullStr iPSC-hepatocyte organoids as a novel platform to predict AAV gene therapy efficacy
title_full_unstemmed iPSC-hepatocyte organoids as a novel platform to predict AAV gene therapy efficacy
title_short iPSC-hepatocyte organoids as a novel platform to predict AAV gene therapy efficacy
title_sort ipsc hepatocyte organoids as a novel platform to predict aav gene therapy efficacy
topic AAV vectors
liver
iPSC
organoids
AAV transduction
AAV serotype
url http://www.sciencedirect.com/science/article/pii/S2329050125000622
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