CRISPR-Mediated Genomic Addition to CPS1 Deficient iPSCs is Insufficient to Restore Nitrogen Homeostasis
| Autor: | Nitzahn M, Truong B, Khoja S, Vega-Crespo A, Le C, Eliav A, Makris G, Ad, Pyle, Häberle J, Gerald Lipshutz |
|---|---|
| Přispěvatelé: | University of Zurich |
| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
iPSC
hyperammonemia Nitrogen Induced Pluripotent Stem Cells Carbamoyl-Phosphate Synthase (Ammonia) 610 Medicine & health Original Contribution Genomics gene therapy Carbamoyl phosphate synthetase deficiency 10036 Medical Clinic Homeostasis Humans Clustered Regularly Interspaced Short Palindromic Repeats CRISPR/Cas9 |
| Zdroj: | The Yale Journal of Biology and Medicine Scopus-Elsevier Europe PubMed Central |
| ISSN: | 1551-4056 0044-0086 |
| Popis: | CPS1 deficiency is an inborn error of metabolism caused by loss-of-function mutations in the CPS1 gene, catalyzing the initial reaction of the urea cycle. Deficiency typically leads to toxic levels of plasma ammonia, cerebral edema, coma, and death, with the only curative treatment being liver transplantation; due to limited donor availability and the invasiveness and complications of the procedure, however, alternative therapies are needed. Induced pluripotent stem cells offer an alternative cell source to partial or whole liver grafts that theoretically would not require immune suppression regimens and additionally are amenable to genetic modifications. Here, we genetically modified CPS1 deficient patient-derived stem cells to constitutively express human codon optimized CPS1 from the AAVS1 safe harbor site. While edited stem cells efficiently differentiated to hepatocyte-like cells, they failed to metabolize ammonia more efficiently than their unedited counterparts. This unexpected result appears to have arisen in part due to transgene promoter methylation, and thus transcriptional silencing, in undifferentiated cells, impacting their capacity to restore the complete urea cycle function upon differentiation. As pluripotent stem cell strategies are being expanded widely for potential cell therapies, these results highlight the need for strict quality control and functional analysis to ensure the integrity of cell products. |
| Databáze: | OpenAIRE |
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