Journal
CELL REPORTS MEDICINE
Volume 2, Issue 4, Pages -Publisher
CELL PRESS
DOI: 10.1016/j.xcrm.2021.100247
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Funding
- National Heart, Lung, and Blood Institute (NHLBI) at NIH
- National Institute of Diabetes, Digestive and Kidney Diseases (NIDDK) at NIH
- National Institute of Allergy and Infectious Diseases (NIAID) at NIH
- Maryland Stem Cell Research Fund (MSCRF) [2015-MSCRFP-1752]
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The study achieved successful gene correction of SCD mutation in CD34+ cells through electroporation and demonstrated engraftment of edited cells in xenograft mice and macaques, providing valuable insights for designing HSC-targeted gene correction trials.
Sickle cell disease (SCD) is caused by a 20A > T mutation in the beta-globin gene. Genome-editing technologies have the potential to correct the SCD mutation in hematopoietic stem cells (HSCs), producing adult hemoglobin while simultaneously eliminating sickle hemoglobin. Here, we developed high-efficiency viral vector-free non-footprint gene correction in SCD CD34+ cells with electroporation to deliver SCD mutation-targeting guide RNA, Cas9 endonuclease, and 100-mer single-strand donor DNA encoding intact beta-globin sequence, achieving therapeutic-level gene correction at DNA (similar to 30%) and protein (similar to 80%) levels. Gene-edited SCD CD34(+) cells contributed corrected cells 6 months post-xenograft mouse transplant without off-target delta-globin editing. We then developed a rhesus beta-to-beta-globin gene conversion strategy to model HSC-targeted genome editing for SCD and demonstrate the engraftment of gene-edited CD34(+) cells 10-12 months post-transplant in rhesus macaques. In summary, gene-corrected CD34(+) HSCs are engraftable in xenograft mice and non-human primates. These findings are helpful in designing HSC-targeted gene correction trials.
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