| RRC ID |
77337
|
| 著者 |
Li C, Psatha N, Sova P, Gil S, Wang H, Kim J, Kulkarni C, Valensisi C, Hawkins RD, Stamatoyannopoulos G, Lieber A.
|
| タイトル |
Reactivation of γ-globin in adult β-YAC mice after ex vivo and in vivo hematopoietic stem cell genome editing.
|
| ジャーナル |
Blood
|
| Abstract |
Disorders involving β-globin gene mutations, primarily β-thalassemia and sickle cell disease, represent a major target for hematopoietic stem/progenitor cell (HSPC) gene therapy. This includes CRISPR/Cas9-mediated genome editing approaches in adult CD34+ cells aimed toward the reactivation of fetal γ-globin expression in red blood cells. Because models involving erythroid differentiation of CD34+ cells have limitations in assessing γ-globin reactivation, we focused on human β-globin locus-transgenic (β-YAC) mice. We used a helper-dependent human CD46-targeting adenovirus vector expressing CRISPR/Cas9 (HDAd-HBG-CRISPR) to disrupt a repressor binding region within the γ-globin promoter. We transduced HSPCs from β-YAC/human CD46-transgenic mice ex vivo and subsequently transplanted them into irradiated recipients. Furthermore, we used an in vivo HSPC transduction approach that involves HSPC mobilization and the intravenous injection of HDAd-HBG-CRISPR into β-YAC/CD46-transgenic mice. In both models, we demonstrated efficient target site disruption, resulting in a pronounced switch from human β- to γ-globin expression in red blood cells of adult mice that was maintained after secondary transplantation of HSPCs. In long-term follow-up studies, we did not detect hematological abnormalities, indicating that HBG promoter editing does not negatively affect hematopoiesis. This is the first study that shows successful in vivo HSPC genome editing by CRISPR/Cas9.
|
| 巻・号 |
131(26)
|
| ページ |
2915-2928
|
| 公開日 |
2018-6-28
|
| DOI |
10.1182/blood-2018-03-838540
|
| PII |
S0006-4971(20)32118-2
|
| PMID |
29789357
|
| PMC |
PMC6024639
|
| MeSH |
Animals
CRISPR-Cas Systems*
Erythrocytes / metabolism
Female
Gene Editing / methods*
Gene Expression*
Hematopoietic Stem Cell Mobilization
Hematopoietic Stem Cell Transplantation
Hematopoietic Stem Cells / metabolism*
Humans
Mice
Mice, Inbred C57BL
Mice, Transgenic
Promoter Regions, Genetic
beta-Globins / genetics*
gamma-Globins / genetics*
|
| IF |
17.794
|
| リソース情報 |
| ヒト・動物細胞 |
HUDEP-2(RCB4557) |
