Highly efficient editing of the β-globin gene in patient-derived hematopoietic stem and progenitor cells to treat sickle cell disease
| Autor: | Gang Bao, Vivien A. Sheehan, Matthew H. Porteus, Alireza Paikari, So Hyun Park, Waracharee Srifa, Yankai Zhang, Alicia K Chang, Timothy H. Davis, Ciaran M. Lee, Joab Camarena, Daniel P. Dever |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
Erythrocytes
medicine.medical_treatment Anemia Sickle Cell Mice SCID beta-Globins Hematopoietic stem cell transplantation Genome Integrity Repair and Replication Biology Mice 03 medical and health sciences 0302 clinical medicine Mice Inbred NOD Cell Line Tumor Genetics medicine Animals Humans Progenitor cell Cells Cultured 030304 developmental biology Gene Editing Mice Knockout 0303 health sciences Hematopoietic Stem Cell Transplantation Genetic Therapy Hematopoietic Stem Cells medicine.disease Sickle cell anemia 3. Good health Transplantation Haematopoiesis Treatment Outcome medicine.anatomical_structure 030220 oncology & carcinogenesis Cancer research Bone marrow CRISPR-Cas Systems Stem cell K562 Cells K562 cells |
| Zdroj: | Nucleic Acids Research |
| ISSN: | 1362-4962 0305-1048 |
| DOI: | 10.1093/nar/gkz475 |
| Popis: | Sickle cell disease (SCD) is a monogenic disorder that affects millions worldwide. Allogeneic hematopoietic stem cell transplantation is the only available cure. Here, we demonstrate the use of CRISPR/Cas9 and a short single-stranded oligonucleotide template to correct the sickle mutation in the β-globin gene in hematopoietic stem and progenitor cells (HSPCs) from peripheral blood or bone marrow of patients with SCD, with 24.5 ± 7.6% efficiency without selection. Erythrocytes derived from gene-edited cells showed a marked reduction of sickle cells, with the level of normal hemoglobin (HbA) increased to 25.3 ± 13.9%. Gene-corrected SCD HSPCs retained the ability to engraft when transplanted into non-obese diabetic (NOD)-SCID-gamma (NSG) mice with detectable levels of gene correction 16–19 weeks post-transplantation. We show that, by using a high-fidelity SpyCas9 that maintained the same level of on-target gene modification, the off-target effects including chromosomal rearrangements were significantly reduced. Taken together, our results demonstrate efficient gene correction of the sickle mutation in both peripheral blood and bone marrow-derived SCD HSPCs, a significant reduction in sickling of red blood cells, engraftment of gene-edited SCD HSPCs in vivo and the importance of reducing off-target effects; all are essential for moving genome editing based SCD treatment into clinical practice. |
| Databáze: | OpenAIRE |
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