| RRC ID |
77341
|
| 著者 |
Psatha N, Georgakopoulou A, Li C, Nandakumar V, Georgolopoulos G, Acosta R, Paschoudi K, Nelson J, Chee D, Athanasiadou A, Kouvatsi A, Funnell APW, Lieber A, Yannaki E, Papayannopoulou T.
|
| タイトル |
Enhanced HbF reactivation by multiplex mutagenesis of thalassemic CD34+ cells in vitro and in vivo.
|
| ジャーナル |
Blood
|
| Abstract |
Thalassemia or sickle cell patients with hereditary persistence of fetal hemoglobin (HbF) have an ameliorated clinical phenotype and, in some cases, can achieve transfusion independence. Inactivation via genome editing of γ-globin developmental suppressors, such as BCL11A or LRF/ZBTB7A, or of their binding sites, have been shown to significantly increase expression of endogenous HbF. To broaden the therapeutic window beyond a single-editing approach, we have explored combinations of cis- and trans-editing targets to enhance HbF reactivation. Multiplex mutagenesis in adult CD34+ cells was well tolerated and did not lead to any detectable defect in the cells' proliferation and differentiation, either in vitro or in vivo. The combination of 1 trans and 1 cis mutation resulted in high editing retention in vivo, coupled with almost pancellular HbF expression in NBSGW mice. The greater in vivo performance of this combination was also recapitulated using a novel helper-dependent adenoviral-CRISPR vector (HD-Ad-dualCRISPR) in CD34+ cells from β-thalassemia patients transplanted to NBSGW mice. A pronounced increase in HbF expression was observed in human red blood cells in mice with established predominant β0/β0-thalassemic hemopoiesis after in vivo injection of the HD-Ad-dualCRISPR vector. Collectively, our data suggest that the combination of cis and trans fetal globin reactivation mutations has the potential to significantly increase HbF both totally and on a per cell basis over single editing and could thus provide significant clinical benefit to patients with severe β-globin phenotype.
|
| 巻・号 |
138(17)
|
| ページ |
1540-1553
|
| 公開日 |
2021-10-28
|
| DOI |
10.1182/blood.2020010020
|
| PII |
S0006-4971(21)01163-0
|
| PMID |
34086867
|
| PMC |
PMC8554647
|
| MeSH |
Adult
Animals
Antigens, CD34 / genetics*
CRISPR-Cas Systems
Cells, Cultured
Fetal Hemoglobin / genetics*
Gene Editing
Genetic Therapy
Humans
Mice
Mutagenesis*
beta-Thalassemia / genetics*
beta-Thalassemia / therapy
gamma-Globins / genetics
|
| IF |
17.794
|
| リソース情報 |
| ヒト・動物細胞 |
HUDEP-2(RCB4557) |
