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Using a barcoded AAV capsid library to select for clinically relevant gene therapy vectors
Katja Pekrun, … , Markus Grompe, Mark A. Kay
Katja Pekrun, … , Markus Grompe, Mark A. Kay
Published November 14, 2019
Citation Information: JCI Insight. 2019;4(22):e131610. https://doi.org/10.1172/jci.insight.131610.
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Resource and Technical Advance Therapeutics

Using a barcoded AAV capsid library to select for clinically relevant gene therapy vectors

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Abstract

While gene transfer using recombinant adeno-associated viral (rAAV) vectors has shown success in some clinical trials, there remain many tissues that are not well transduced. Because of the recent success in reprogramming islet-derived cells into functional β cells in animal models, we constructed 2 highly complex barcoded replication competent capsid shuffled libraries and selected for high-transducing variants on primary human islets. We describe the generation of a chimeric AAV capsid (AAV-KP1) that facilitates transduction of primary human islet cells and human embryonic stem cell–derived β cells with up to 10-fold higher efficiency compared with previously studied best-in-class AAV vectors. Remarkably, this chimeric capsid also enabled transduction of both mouse and human hepatocytes at very high levels in a humanized chimeric mouse model, thus providing a versatile vector that has the potential to be used in both preclinical testing and human clinical trials for liver-based diseases and diabetes.

Authors

Katja Pekrun, Gustavo De Alencastro, Qing-Jun Luo, Jun Liu, Youngjin Kim, Sean Nygaard, Feorillo Galivo, Feijie Zhang, Ren Song, Matthew R. Tiffany, Jianpeng Xu, Matthias Hebrok, Markus Grompe, Mark A. Kay

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Figure 1

Evaluation of parental capsids for islet tropism.

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Evaluation of parental capsids for islet tropism.
(A) Dissociated islet ...
(A) Dissociated islet cells were transduced with different vectors that had been packaged with AAV2, AAV3B, DJ, and LK03 capsids at a MOI of 1,000, and transduction efficiency was determined by flow cytometry 48 hours after transduction. Each rAAV was tested once. (B) Parental contribution in the 18-parent pool as analyzed using high-throughput sequencing of the barcodes. (C) Barcode sequences were amplified from viral genomes after passaging the 18-parent pool on human islets and were analyzed by high-throughput sequencing. Color coding of enriched parental capsids for passaging of the 18-parent pool is identical to that of the input parental pool.

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ISSN 2379-3708

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