دورية أكاديمية
Ex vivo gene editing and cell therapy for hereditary tyrosinemia type 1
| العنوان: | Ex vivo gene editing and cell therapy for hereditary tyrosinemia type 1 |
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| المؤلفون: | Ilayda Ates, Callie Stuart, Tanner Rathbone, Mercedes Barzi, Gordon He, Angela M. Major, Vijay Shankar, Rachel A. Lyman, Sidney S. Angner, Trudy F.C. Mackay, Shanthi Srinivasan, Alton Brad Farris, Karl-Dimiter Bissig, Renee N. Cottle |
| المصدر: | Hepatology Communications, Vol 8, Iss 5 (2024) |
| بيانات النشر: | Wolters Kluwer Health/LWW, 2024. |
| سنة النشر: | 2024 |
| المجموعة: | LCC:Diseases of the digestive system. Gastroenterology |
| مصطلحات موضوعية: | Diseases of the digestive system. Gastroenterology, RC799-869 |
| الوصف: | Background:. We previously demonstrated the successful use of in vivo CRISPR gene editing to delete 4-hydroxyphenylpyruvate dioxygenase (HPD) to rescue mice deficient in fumarylacetoacetate hydrolase (FAH), a disorder known as hereditary tyrosinemia type 1 (HT1). The aim of this study was to develop an ex vivo gene-editing protocol and apply it as a cell therapy for HT1. Methods:. We isolated hepatocytes from wild-type (C57BL/6J) and Fah −/− mice and then used an optimized electroporation protocol to deliver Hpd-targeting CRISPR-Cas9 ribonucleoproteins into hepatocytes. Next, hepatocytes were transiently incubated in cytokine recovery media formulated to block apoptosis, followed by splenic injection into recipient Fah −/− mice. Results:. We observed robust engraftment and expansion of transplanted gene-edited hepatocytes from wild-type donors in the livers of recipient mice when transient incubation with our cytokine recovery media was used after electroporation and negligible engraftment without the media (mean: 46.8% and 0.83%, respectively; p=0.0025). Thus, the cytokine recovery medium was critical to our electroporation protocol. When hepatocytes from Fah −/− mice were used as donors for transplantation, we observed 35% and 28% engraftment for Hpd-Cas9 ribonucleoproteins and Cas9 mRNA, respectively. Tyrosine, phenylalanine, and biochemical markers of liver injury normalized in both Hpd-targeting Cas9 ribonucleoprotein and mRNA groups independent of induced inhibition of Hpd through nitisinone, indicating correction of disease indicators in Fah −/− mice. Conclusions:. The successful liver cell therapy for HT1 validates our protocol and, despite the known growth advantage of HT1, showcases ex vivo gene editing using electroporation in combination with liver cell therapy to cure a disease model. These advancements underscore the potential impacts of electroporation combined with transplantation as a cell therapy. |
| نوع الوثيقة: | article |
| وصف الملف: | electronic resource |
| اللغة: | English |
| تدمد: | 2471-254X 00000000 |
| العلاقة: | http://journals.lww.com/10.1097/HC9.0000000000000424Test; https://doaj.org/toc/2471-254XTest |
| DOI: | 10.1097/HC9.0000000000000424 |
| الوصول الحر: | https://doaj.org/article/bfe468ab97a549e5a155fe6c7c05eae7Test |
| رقم الانضمام: | edsdoj.bfe468ab97a549e5a155fe6c7c05eae7 |
| قاعدة البيانات: | Directory of Open Access Journals |
Full Text Finder | تدمد: | 2471254X 00000000 |
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| DOI: | 10.1097/HC9.0000000000000424 |