BCL11A enhancer–edited hematopoietic stem cells persist in rhesus monkeys without toxicity
| Autor: | Yuxuan Wu, Tina Nassehi, Theresa Engels, John F. Tisdale, Aylin C. Bonifacino, Shengdar Q. Tsai, Alexis Leonard, Nathaniel S. Linde, Kevin Luk, Cicera R. Lazzarotto, Robert E. Donahue, Jackson Gamer, Claire M. Drysdale, Jing Zeng, Morgan Yapundich, Selami Demirci, Daniel E. Bauer, Mitchell J. Weiss, Anne H. Shen, Juan J. Haro-Mora, Scot A. Wolfe, Jasmine Bonanno, Allen E. Krouse, Danilo Pellin, Naoya Uchida, Shondra M. Pruett-Miller, Shaina N. Porter |
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| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Biology Transplantation Autologous Mice 03 medical and health sciences 0302 clinical medicine medicine Animals Humans Progenitor cell Enhancer Gene Editing Hematopoietic Stem Cell Transplantation GATA1 General Medicine Hematopoietic Stem Cells Macaca mulatta Molecular biology Repressor Proteins Transplantation Haematopoiesis 030104 developmental biology medicine.anatomical_structure 030220 oncology & carcinogenesis Erythropoiesis Bone marrow Stem cell Research Article |
| Zdroj: | J Clin Invest |
| ISSN: | 1558-8238 0021-9738 |
| Popis: | Gene editing of the erythroid-specific BCL11A enhancer in hematopoietic stem and progenitor cells (HSPCs) from patients with sickle cell disease (SCD) induces fetal hemoglobin (HbF) without detectable toxicity, as assessed by mouse xenotransplant. Here, we evaluated autologous engraftment and HbF induction potential of erythroid-specific BCL11A enhancer–edited HSPCs in 4 nonhuman primates. We used a single guide RNA (sgRNA) with identical human and rhesus target sequences to disrupt a GATA1 binding site at the BCL11A +58 erythroid enhancer. Cas9 protein and sgRNA ribonucleoprotein complex (RNP) was electroporated into rhesus HSPCs, followed by autologous infusion after myeloablation. We found that gene edits persisted in peripheral blood (PB) and bone marrow (BM) for up to 101 weeks similarly for BCL11A enhancer– or control locus–targeted (AAVS1-targeted) cells. Biallelic BCL11A enhancer editing resulted in robust γ-globin induction, with the highest levels observed during stress erythropoiesis. Indels were evenly distributed across PB and BM lineages. Off-target edits were not observed. Nonhomologous end-joining repair alleles were enriched in engrafting HSCs. In summary, we found that edited HSCs can persist for at least 101 weeks after transplant and biallelic-edited HSCs provide substantial HbF levels in PB red blood cells, together supporting further clinical translation of this approach. |
| Databáze: | OpenAIRE |
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