Human iPSC Modeling Reveals Mutation-Specific Responses to Gene Therapy in a Genotypically Diverse Dominant Maculopathy

Divya Sinha; Edwin M Stone; Benjamin Steyer; Budd A Tucker; Pawan K Shahi; Katherine P Mueller; Rasa Valiauga; Kimberly L Edwards; Cole Bacig; Stephanie S Steltzer; Sandhya Srinivasan; Amr Abdeen; Evan Cory; Viswesh Periyasamy; Alireza Fotuhi Siahpirani; Sushmita Roy; Bikash R Pattnaik; Krishanu Saha; David M Gamm

doi:10.1016/j.ajhg.2020.06.011

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Human iPSC Modeling Reveals Mutation-Specific Responses to Gene Therapy in a Genotypically Diverse Dominant Maculopathy

Journal article

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Human iPSC Modeling Reveals Mutation-Specific Responses to Gene Therapy in a Genotypically Diverse Dominant Maculopathy

Divya Sinha, Edwin M Stone, Benjamin Steyer, Budd A Tucker, Pawan K Shahi, Katherine P Mueller, Rasa Valiauga, Kimberly L Edwards, Cole Bacig, Stephanie S Steltzer, …

American journal of human genetics, Vol.107(2), pp.278-292

08/06/2020

DOI: 10.1016/j.ajhg.2020.06.011

PMID: 32707085

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Abstract

Dominantly inherited disorders are not typically considered to be therapeutic candidates for gene augmentation. Here, we utilized induced pluripotent stem cell-derived retinal pigment epithelium (iPSC-RPE) to test the potential of gene augmentation to treat Best disease, a dominant macular dystrophy caused by over 200 missense mutations in BEST1. Gene augmentation in iPSC-RPE fully restored BEST1 calcium-activated chloride channel activity and improved rhodopsin degradation in an iPSC-RPE model of recessive bestrophinopathy as well as in two models of dominant Best disease caused by different mutations in regions encoding ion-binding domains. A third dominant Best disease iPSC-RPE model did not respond to gene augmentation, but showed normalization of BEST1 channel activity following CRISPR-Cas9 editing of the mutant allele. We then subjected all three dominant Best disease iPSC-RPE models to gene editing, which produced premature stop codons specifically within the mutant BEST1 alleles. Single-cell profiling demonstrated no adverse perturbation of retinal pigment epithelium (RPE) transcriptional programs in any model, although off-target analysis detected a silent genomic alteration in one model. These results suggest that gene augmentation is a viable first-line approach for some individuals with dominant Best disease and that non-responders are candidates for alternate approaches such as gene editing. However, testing gene editing strategies for on-target efficiency and off-target events using personalized iPSC-RPE model systems is warranted. In summary, personalized iPSC-RPE models can be used to select among a growing list of gene therapy options to maximize safety and efficacy while minimizing time and cost. Similar scenarios likely exist for other genotypically diverse channelopathies, expanding the therapeutic landscape for affected individuals.

Gene Therapy

somatic cell gene editing

orphan disease

channelopathy

CRISPR-Cas9

gene augmentation

human pluripotent stem cells

autosomal dominant disease

macular degeneration

Details

Title: Subtitle: Human iPSC Modeling Reveals Mutation-Specific Responses to Gene Therapy in a Genotypically Diverse Dominant Maculopathy
Creators: Divya Sinha - McPherson Eye Research Institute, University of Wisconsin—Madison, Madison, WI 53705, USA
Edwin M Stone - Department of Ophthalmology and Visual Sciences, Carver College of Medicine, University of Iowa, Iowa City, IA 52242, USA
Benjamin Steyer - McPherson Eye Research Institute, University of Wisconsin—Madison, Madison, WI 53705, USA
Budd A Tucker - Department of Ophthalmology and Visual Sciences, Carver College of Medicine, University of Iowa, Iowa City, IA 52242, USA
Pawan K Shahi - McPherson Eye Research Institute, University of Wisconsin—Madison, Madison, WI 53705, USA
Katherine P Mueller - Wisconsin Institute for Discovery, University of Wisconsin—Madison, Madison, WI 53715, USA
Rasa Valiauga - Waisman Center, University of Wisconsin—Madison, Madison, WI 53705, USA
Kimberly L Edwards - Waisman Center, University of Wisconsin—Madison, Madison, WI 53705, USA
Cole Bacig - Waisman Center, University of Wisconsin—Madison, Madison, WI 53705, USA
Stephanie S Steltzer - Wisconsin Institute for Discovery, University of Wisconsin—Madison, Madison, WI 53715, USA
Sandhya Srinivasan - Wisconsin Institute for Discovery, University of Wisconsin—Madison, Madison, WI 53715, USA
Amr Abdeen - Wisconsin Institute for Discovery, University of Wisconsin—Madison, Madison, WI 53715, USA
Evan Cory - Wisconsin Institute for Discovery, University of Wisconsin—Madison, Madison, WI 53715, USA
Viswesh Periyasamy - Wisconsin Institute for Discovery, University of Wisconsin—Madison, Madison, WI 53715, USA
Alireza Fotuhi Siahpirani - Wisconsin Institute for Discovery, University of Wisconsin—Madison, Madison, WI 53715, USA
Sushmita Roy - Wisconsin Institute for Discovery, University of Wisconsin—Madison, Madison, WI 53715, USA
Bikash R Pattnaik - McPherson Eye Research Institute, University of Wisconsin—Madison, Madison, WI 53705, USA
Krishanu Saha - McPherson Eye Research Institute, University of Wisconsin—Madison, Madison, WI 53705, USA
David M Gamm - McPherson Eye Research Institute, University of Wisconsin—Madison, Madison, WI 53705, USA
Resource Type: Journal article
Publication Details: American journal of human genetics, Vol.107(2), pp.278-292
Publisher: Elsevier Inc
DOI: 10.1016/j.ajhg.2020.06.011
PMID: 32707085
ISSN: 0002-9297
eISSN: 1537-6605
Grant note: DOI: 10.13039/100001116, name: Foundation Fighting Blindness; DOI: 10.13039/100000001, name: National Science Foundation; DOI: 10.13039/100000002, name: National Institutes of Health
Language: English
Date published: 08/06/2020
Academic Unit: Iowa Neuroscience Institute; Ophthalmology and Visual Sciences
Record Identifier: 9984070774602771

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