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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 8

Validation and quantification of human hepatocyte transduction in mice with humanized liver in vivo.

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Validation and quantification of human hepatocyte transduction in mice w...
(A) Representative immunofluorescence images from livers of mice injected with ssAAV-Td Tomato Red at 1 × 1011 vg i.v. with varying capsid serotypes (3 mice for DJ; 4 mice each for LK03 and KP1). DAPI (blue), human-specific FAH (green), and Tomato Red (red) on liver sections. Scale bar: 50 μM. (B) Quantification of human hepatocyte repopulation levels, transduction efficiency for all hepatocytes, and transduction efficiency for human hepatocytes only. Each data point represents an area of interest for each mouse. A total of 6–9 areas of interest for each mouse were scanned and analyzed. The mean and SD for each mouse is indicated. Experimental values were assessed via 2-way ANOVA using Tukey’s multiple comparisons test. Only statistically significant differences between the groups are shown in the graphs. **P < 0.01, ****P < 0.000.

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