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dc.contributor.authorDemirci, Selami
dc.contributor.authorZeng, Jing
dc.contributor.authorWu, Yuxuan
dc.contributor.authorUchida, Naoya
dc.contributor.authorShen, Anne H.
dc.contributor.authorPellin, Danilo
dc.contributor.authorGamer, Jackson
dc.contributor.authorYapundich, Morgan
dc.contributor.authorDrysdale, Claire
dc.contributor.authorBonanno, Jasmine
dc.contributor.authorBonifacino, Aylin C.
dc.contributor.authorKrouse, Allen
dc.contributor.authorLinde, Nathaniel Seth.
dc.contributor.authorEngels, Theresa
dc.contributor.authorDonahue, Robert E.
dc.contributor.authorHaro-Mora, Juan J.
dc.contributor.authorLeonard, Alexis
dc.contributor.authorNassehi, Tina
dc.contributor.authorLuk, Kevin
dc.contributor.authorPorter, Shaina N.
dc.contributor.authorLazzarotto, Cicera R.
dc.contributor.authorTsai, Shengdar Q.
dc.contributor.authorWeiss, Mitchell
dc.contributor.authorPruett-Miller, Shondra M.
dc.contributor.authorWolfe, Scot A.
dc.contributor.authorBauer, Daniel E.
dc.contributor.authorTisdale, John F.
dc.date2022-08-11T08:09:56.000
dc.date.accessioned2022-08-23T16:50:00Z
dc.date.available2022-08-23T16:50:00Z
dc.date.issued2020-09-08
dc.date.submitted2020-10-26
dc.identifier.citation<p>Demirci S, Zeng J, Wu Y, Uchida N, Shen AH, Pellin D, Gamer J, Yapundich M, Drysdale C, Bonanno J, Bonifacino AC, Krouse A, Linde NS, Engels T, Donahue RE, Haro-Mora JJ, Leonard A, Nassehi T, Luk K, Porter SN, Lazzarotto CR, Tsai SQ, Weiss M, Pruett-Miller SM, Wolfe SA, Bauer DE, Tisdale JF. BCL11A enhancer edited hematopoietic stem cells persist in rhesus monkeys without toxicity. J Clin Invest. 2020 Sep 8:140189. doi: 10.1172/JCI140189. Epub ahead of print. PMID: 32897878. <a href="https://doi.org/10.1172/JCI140189">Link to article on publisher's site</a></p>
dc.identifier.issn0021-9738 (Linking)
dc.identifier.doi10.1172/JCI140189
dc.identifier.pmid32897878
dc.identifier.urihttp://hdl.handle.net/20.500.14038/41575
dc.description.abstractGene editing of the erythroid-specific BCL11A enhancer in hematopoietic stem and progenitor cells (HSPCs) from sickle cell disease (SCD) patients 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 four non-human primates. We utilized 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 (AAVS1) targeted cells. Biallelic BCL11A enhancer editing resulted in robust gamma-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. Non-homologous end-joining repair alleles were enriched in engrafting HSCs. In summary, we find that edited HSCs can persist for at least 101 weeks post-transplant, and biallelic edited HSCs provide substantial HbF levels in PB red blood cells, together supporting further clinical translation of this approach.
dc.language.isoen_US
dc.relation<p><a href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=pubmed&cmd=Retrieve&list_uids=32897878&dopt=Abstract">Link to Article in PubMed</a></p>
dc.relation.urlhttps://doi.org/10.1172/jci140189
dc.rightsCopyright © 2020 American Society for Clinical Investigation. Authors' accepted manuscript posted as allowed by the publisher's self-archiving policy at https://v2.sherpa.ac.uk/id/publication/6211.
dc.subjectGenetic diseases
dc.subjectHematology
dc.subjectHematopoietic stem cells
dc.subjectStem cell transplantation
dc.subjectTransplantation
dc.subjectSickle cell disease
dc.subjectβ-thalassemia
dc.subjectγ-globin
dc.subjectCRISPR/Cas9
dc.subjectgene editing
dc.subjectAmino Acids, Peptides, and Proteins
dc.subjectCell Biology
dc.subjectCongenital, Hereditary, and Neonatal Diseases and Abnormalities
dc.subjectGenetics and Genomics
dc.subjectHematology
dc.subjectHemic and Lymphatic Diseases
dc.subjectMedical Genetics
dc.subjectNucleic Acids, Nucleotides, and Nucleosides
dc.titleBCL11A enhancer edited hematopoietic stem cells persist in rhesus monkeys without toxicity
dc.typeAccepted Manuscript
dc.source.journaltitleThe Journal of clinical investigation
dc.identifier.legacyfulltexthttps://escholarship.umassmed.edu/cgi/viewcontent.cgi?article=5393&amp;context=oapubs&amp;unstamped=1
dc.identifier.legacycoverpagehttps://escholarship.umassmed.edu/oapubs/4365
dc.identifier.contextkey19982993
refterms.dateFOA2022-08-23T16:50:00Z
html.description.abstract<p>Gene editing of the erythroid-specific BCL11A enhancer in hematopoietic stem and progenitor cells (HSPCs) from sickle cell disease (SCD) patients 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 four non-human primates. We utilized 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 (AAVS1) targeted cells. Biallelic BCL11A enhancer editing resulted in robust gamma-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. Non-homologous end-joining repair alleles were enriched in engrafting HSCs. In summary, we find that edited HSCs can persist for at least 101 weeks post-transplant, and biallelic edited HSCs provide substantial HbF levels in PB red blood cells, together supporting further clinical translation of this approach.</p>
dc.identifier.submissionpathoapubs/4365
dc.contributor.departmentGraduate School of Biomedical Sciences


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