| Autor: |
Carlson-Stevemer, Jared, Das, Amritava, Fiflis, David Nicholas, Abdeen, Amr, Kleitzen, Heidi, Kohlenberg, Lucille, Goedland, Madelyn, Connor, Nadine, Saha, Krishanu |
| Jazyk: |
angličtina |
| Rok vydání: |
2019 |
| Předmět: |
|
| DOI: |
10.5281/zenodo.2556885 |
| Popis: |
Genome editing has high potential to directly correct endogenous diseased alleles in the body for the treatment of many inherited disorders. While potential off-target effects and other safety concerns have been extensively studied, the efficacy of genome editing strategies have yet to be quantitatively analyzed, especially in cases where no suitable animal model exists. Furthermore, genome editors have been engineered to be more precise, opening up new strategies for editing multiple alleles in polygenic cases. Here, we develop experimental and mathematical models for the efficacy and potency of rescuing the glycogen storage diseased phenotype in Pompe disease with CRISPR-Cas9 genome editing. Using a patient-derived induced pluripotent stem cell (iPSC) model of Pompe disease, we demonstrate that gene correction of two mutant alleles within an autosomal recessive case of Pompe disease rescues the diseased phenotype more effectively than correcting a single mutant allele. Edited cells had normal karyotypes, no detectable off-target mutations or large genetic deletions, and lower amounts of abnormal glycogen storage. Enzymatic cross-correction of the diseased cells from GAA produced within edited cells was robust, and this enzymatic cross-correction was equivalent to standard enzyme replacement therapy (ERT). Mathematical modeling for the in vivo delivery of these genome editors to the livers of Pompe patients confirmed the efficacy of disease rescue for precise genome editors with six serial doses, as compared to long-term biweekly ERT. Progenitor cell targeting, delivery efficiencies, and suppression of imprecise editing outcomes were identified as key parameters controlling efficacy. This work establishes new quantitative design rules for somatic cell genome editing, and that precise correction using serial dosing of genome editors to correct multiple mutant alleles may be efficacious in the treatment of polygenic disorders. |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
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