Cited 27 times in
Curative Ex Vivo Hepatocyte-Directed Gene Editing in a Mouse Model of Hereditary Tyrosinemia Type 1
DC Field | Value | Language |
---|---|---|
dc.contributor.author | 주동진 | - |
dc.date.accessioned | 2019-12-18T00:19:45Z | - |
dc.date.available | 2019-12-18T00:19:45Z | - |
dc.date.issued | 2018 | - |
dc.identifier.issn | 1043-0342 | - |
dc.identifier.uri | https://ir.ymlib.yonsei.ac.kr/handle/22282913/172980 | - |
dc.description.abstract | Hereditary tyrosinemia type 1 (HT1) is an autosomal recessive disorder caused by deficiency of fumarylacetoacetate hydrolase (FAH). It has been previously shown that ex vivo hepatocyte-directed gene therapy using an integrating lentiviral vector to replace the defective Fah gene can cure liver disease in small- and large-animal models of HT1. This study hypothesized that ex vivo hepatocyte-directed gene editing using CRISPR/Cas9 could be used to correct a mouse model of HT1, in which a single point mutation results in loss of FAH function. To achieve high transduction efficiencies of primary hepatocytes, this study utilized a lentiviral vector (LV) to deliver both the Streptococcus pyogenes Cas9 nuclease and target guide RNA (LV-Cas9) and an adeno-associated virus (AAV) vector to deliver a 1.2 kb homology template (AAV-HT). Cells were isolated from Fah-/- mice and cultured in the presence of LV and AAV vectors. Transduction of cells with LV-Cas9 induced significant indels at the target locus, and correction of the point mutation in Fah-/- cells ex vivo using AAV-HT was completely dependent on LV-Cas9. Next, hepatocytes transduced ex vivo by LV-Cas9 and AAV-HT were transplanted into syngeneic Fah-/- mice that had undergone a two-thirds partial hepatectomy or sham hepatectomy. Mice were cycled on/off the protective drug 2-(2-nitro-4-trifluoromethylbenzoyl)-1,3-cyclohexanedione (NTBC) to stimulate expansion of corrected cells. All transplanted mice became weight stable off NTBC. However, a significant improvement was observed in weight stability off NTBC in animals that received partial hepatectomy. After 6 months, mice were euthanized, and thorough biochemical and histological examinations were performed. Biochemical markers of liver injury were significantly improved over non-transplanted controls. Histological examination of mice revealed normal tissue architecture, while immunohistochemistry showed robust repopulation of recipient animals with FAH+ cells. In summary, this is the first report of ex vivo hepatocyte-directed gene repair using CRISPR/Cas9 to demonstrate curative therapy in an animal model of liver disease. | - |
dc.description.statementOfResponsibility | open | - |
dc.language | English | - |
dc.publisher | M.A. Liebert | - |
dc.relation.isPartOf | HUMAN GENE THERAPY | - |
dc.rights | CC BY-NC-ND 2.0 KR | - |
dc.subject.MESH | Animals | - |
dc.subject.MESH | Base Sequence | - |
dc.subject.MESH | CRISPR-Associated Protein 9/metabolism | - |
dc.subject.MESH | Cells, Cultured | - |
dc.subject.MESH | Clustered Regularly Interspaced Short Palindromic Repeats/genetics | - |
dc.subject.MESH | Dependovirus/metabolism | - |
dc.subject.MESH | Disease Models, Animal | - |
dc.subject.MESH | Gene Editing* | - |
dc.subject.MESH | Genetic Therapy* | - |
dc.subject.MESH | Genetic Vectors/metabolism | - |
dc.subject.MESH | Hepatocytes/metabolism* | - |
dc.subject.MESH | Hepatocytes/transplantation | - |
dc.subject.MESH | Hydrolases/genetics | - |
dc.subject.MESH | Lentivirus/genetics | - |
dc.subject.MESH | Liver Failure/pathology | - |
dc.subject.MESH | Liver Failure/therapy | - |
dc.subject.MESH | Mice | - |
dc.subject.MESH | Tyrosinemias/genetics* | - |
dc.subject.MESH | Tyrosinemias/pathology | - |
dc.subject.MESH | Tyrosinemias/therapy* | - |
dc.title | Curative Ex Vivo Hepatocyte-Directed Gene Editing in a Mouse Model of Hereditary Tyrosinemia Type 1 | - |
dc.type | Article | - |
dc.contributor.college | College of Medicine (의과대학) | - |
dc.contributor.department | Dept. of Surgery (외과학교실) | - |
dc.contributor.googleauthor | Caitlin VanLith | - |
dc.contributor.googleauthor | Rebekah Guthman | - |
dc.contributor.googleauthor | Clara T. Nicolas | - |
dc.contributor.googleauthor | Kari Allen | - |
dc.contributor.googleauthor | Zeji Du | - |
dc.contributor.googleauthor | Dong Jin Joo | - |
dc.contributor.googleauthor | Scott L. Nyberg | - |
dc.contributor.googleauthor | Joseph B. Lillegard | - |
dc.contributor.googleauthor | Raymond D. Hickey | - |
dc.identifier.doi | 10.1089/hum.2017.252 | - |
dc.contributor.localId | A03948 | - |
dc.relation.journalcode | J01006 | - |
dc.identifier.eissn | 1557-7422 | - |
dc.identifier.pmid | 29764210 | - |
dc.subject.keyword | CRISPR/Cas9 | - |
dc.subject.keyword | gene therapy | - |
dc.subject.keyword | hepatocytes | - |
dc.subject.keyword | hereditary tyrosinemia type 1 | - |
dc.subject.keyword | metabolic liver disease | - |
dc.contributor.alternativeName | Joo, Dong Jin | - |
dc.contributor.affiliatedAuthor | 주동진 | - |
dc.citation.volume | 29 | - |
dc.citation.number | 11 | - |
dc.citation.startPage | 1315 | - |
dc.citation.endPage | 1326 | - |
dc.identifier.bibliographicCitation | HUMAN GENE THERAPY, Vol.29(11) : 1315-1326, 2018 | - |
Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.