Comparative targeting analysis of KLF1, BCL11A, and HBG1/2 in CD34
Anemia, Sickle Cell
/ genetics
Antigens, CD34
CRISPR-Cas Systems
Cells, Cultured
Fetal Hemoglobin
/ genetics
Gene Editing
/ methods
Gene Expression
/ genetics
Humans
Kruppel-Like Transcription Factors
/ genetics
Molecular Targeted Therapy
Mutation
Repressor Proteins
/ genetics
gamma-Globins
/ genetics
Journal
Scientific reports
ISSN: 2045-2322
Titre abrégé: Sci Rep
Pays: England
ID NLM: 101563288
Informations de publication
Date de publication:
23 06 2020
23 06 2020
Historique:
received:
18
10
2019
accepted:
19
05
2020
entrez:
25
6
2020
pubmed:
25
6
2020
medline:
16
1
2021
Statut:
epublish
Résumé
β-hemoglobinopathies are caused by abnormal or absent production of hemoglobin in the blood due to mutations in the β-globin gene (HBB). Imbalanced expression of adult hemoglobin (HbA) induces strong anemia in patients suffering from the disease. However, individuals with natural-occurring mutations in the HBB cluster or related genes, compensate this disparity through γ-globin expression and subsequent fetal hemoglobin (HbF) production. Several preclinical and clinical studies have been performed in order to induce HbF by knocking-down genes involved in HbF repression (KLF1 and BCL11A) or disrupting the binding sites of several transcription factors in the γ-globin gene (HBG1/2). In this study, we thoroughly compared the different CRISPR/Cas9 gene-disruption strategies by gene editing analysis and assessed their safety profile by RNA-seq and GUIDE-seq. All approaches reached therapeutic levels of HbF after gene editing and showed similar gene expression to the control sample, while no significant off-targets were detected by GUIDE-seq. Likewise, all three gene editing platforms were established in the GMP-grade CliniMACS Prodigy, achieving similar outcome to preclinical devices. Based on this gene editing comparative analysis, we concluded that BCL11A is the most clinically relevant approach while HBG1/2 could represent a promising alternative for the treatment of β-hemoglobinopathies.
Identifiants
pubmed: 32576837
doi: 10.1038/s41598-020-66309-x
pii: 10.1038/s41598-020-66309-x
pmc: PMC7311455
doi:
Substances chimiques
Antigens, CD34
0
BCL11A protein, human
0
Kruppel-Like Transcription Factors
0
Repressor Proteins
0
erythroid Kruppel-like factor
0
gamma-Globins
0
Fetal Hemoglobin
9034-63-3
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
10133Références
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