用β珠蛋白基因替换α珠蛋白可恢复β地中海贫血源性造血干细胞和祖细胞中的血红蛋白平衡。
Gene replacement of α-globin with β-globin restores hemoglobin balance in β-thalassemia-derived hematopoietic stem and progenitor cells.
机构信息
Department of Pediatrics, Stanford University, Stanford, CA, USA.
Integrated DNA Technologies, Inc., Coralville, IA, USA.
出版信息
Nat Med. 2021 Apr;27(4):677-687. doi: 10.1038/s41591-021-01284-y. Epub 2021 Mar 18.
Abstract
β-Thalassemia pathology is due not only to loss of β-globin (HBB), but also to erythrotoxic accumulation and aggregation of the β-globin-binding partner, α-globin (HBA1/2). Here we describe a Cas9/AAV6-mediated genome editing strategy that can replace the entire HBA1 gene with a full-length HBB transgene in β-thalassemia-derived hematopoietic stem and progenitor cells (HSPCs), which is sufficient to normalize β-globin:α-globin messenger RNA and protein ratios and restore functional adult hemoglobin tetramers in patient-derived red blood cells. Edited HSPCs were capable of long-term and bilineage hematopoietic reconstitution in mice, establishing proof of concept for replacement of HBA1 with HBB as a novel therapeutic strategy for curing β-thalassemia.
摘要
β-地中海贫血的病理不仅源于β-球蛋白（HBB）的缺失，还源于毒性红系细胞内β-球蛋白结合伴侣α-球蛋白（HBA1/2）的积累和聚集。在此，我们描述了一种 Cas9/AAV6 介导的基因组编辑策略，它可以将 HBA1 基因的全长替换为β-地中海贫血衍生造血干细胞和祖细胞（HSPCs）中的全长 HBB 转基因，足以使β-球蛋白：α-球蛋白信使 RNA 和蛋白质的比值正常化，并在患者来源的红细胞中恢复功能性成人血红蛋白四聚体。编辑后的 HSPCs 能够在小鼠中进行长期的双谱系造血重建，为用 HBB 替代 HBA1 作为治疗β-地中海贫血的新策略提供了概念验证。
相似文献
1
Gene replacement of α-globin with β-globin restores hemoglobin balance in β-thalassemia-derived hematopoietic stem and progenitor cells.用β珠蛋白基因替换α珠蛋白可恢复β地中海贫血源性造血干细胞和祖细胞中的血红蛋白平衡。
Nat Med. 2021 Apr;27(4):677-687. doi: 10.1038/s41591-021-01284-y. Epub 2021 Mar 18.
2
Correction of β-thalassemia by CRISPR/Cas9 editing of the α-globin locus in human hematopoietic stem cells.通过在人造血干细胞中编辑α-珠蛋白基因座对β-地中海贫血的校正。
Blood Adv. 2021 Mar 9;5(5):1137-1153. doi: 10.1182/bloodadvances.2020001996.
3
CRISPR/Cas9 β-globin gene targeting in human haematopoietic stem cells.CRISPR/Cas9对人类造血干细胞β-珠蛋白基因的靶向作用。
Nature. 2016 Nov 17;539(7629):384-389. doi: 10.1038/nature20134. Epub 2016 Nov 7.
4
Editing an α-globin enhancer in primary human hematopoietic stem cells as a treatment for β-thalassemia.编辑原代人类造血干细胞中的α-珠蛋白增强子作为β-地中海贫血的一种治疗方法。
Nat Commun. 2017 Sep 4;8(1):424. doi: 10.1038/s41467-017-00479-7.
5
Base editing of haematopoietic stem cells rescues sickle cell disease in mice.碱基编辑造血干细胞可拯救患镰状细胞病的小鼠。
Nature. 2021 Jul;595(7866):295-302. doi: 10.1038/s41586-021-03609-w. Epub 2021 Jun 2.
6
Genome Editing for the β-Hemoglobinopathies.用于β地中海贫血的基因组编辑
Adv Exp Med Biol. 2017;1013:203-217. doi: 10.1007/978-1-4939-7299-9_8.
7
Reactivation of γ-globin in adult β-YAC mice after ex vivo and in vivo hematopoietic stem cell genome editing.经体外和体内造血干细胞基因组编辑后，β-YAC 小鼠成年后 γ-珠蛋白的重新激活。
Blood. 2018 Jun 28;131(26):2915-2928. doi: 10.1182/blood-2018-03-838540. Epub 2018 May 22.
8
Recombinant AAV2-mediated β-globin expression in human fetal hematopoietic cells from the aborted fetuses with β-thalassemia major.经重组 AAV2 介导的β-珠蛋白在患有重型β-地中海贫血症的流产胎儿人胎儿造血细胞中的表达。
Int J Hematol. 2011 Jun;93(6):691-699. doi: 10.1007/s12185-011-0823-x. Epub 2011 May 27.
9
Comparative analysis of FV vectors with human α- or β-globin gene regulatory elements for the correction of β-thalassemia.比较分析含有人α或β珠蛋白基因调控元件的 FV 载体，用于β-地中海贫血的矫正。
Gene Ther. 2012 Mar;19(3):303-11. doi: 10.1038/gt.2011.98. Epub 2011 Jul 7.
10
Hematopoietic Stem Cell Gene-Addition/Editing Therapy in Sickle Cell Disease.造血干细胞基因添加/编辑治疗镰状细胞病。
Cells. 2022 Jun 4;11(11):1843. doi: 10.3390/cells11111843.
引用本文的文献
1
Gene editing in hematopoietic stem cells by co-delivery of Cas9/sgRNA ribonucleoprotein and templates for homology-directed repair in 'all-in-one' lentivirus-derived nanoparticles.通过在“一体化”慢病毒衍生纳米颗粒中共递送Cas9/sgRNA核糖核蛋白和同源定向修复模板，对造血干细胞进行基因编辑。
Nucleic Acids Res. 2025 Aug 11;53(15). doi: 10.1093/nar/gkaf767.
2
Targeted gene editing and near-universal cDNA insertion of CYBA and CYBB as a treatment for chronic granulomatous disease.靶向基因编辑以及CYBA和CYBB的近乎普遍的cDNA插入作为慢性肉芽肿病的一种治疗方法。
Nat Commun. 2025 Aug 12;16(1):7475. doi: 10.1038/s41467-025-62738-2.
3
Protocol for efficient CRISPR/AAV-mediated genome editing and erythroid differentiation of human hematopoietic stem and progenitor cells.高效CRISPR/AAV介导的人类造血干细胞和祖细胞基因组编辑及红系分化方案。
STAR Protoc. 2025 Aug 5;6(3):104018. doi: 10.1016/j.xpro.2025.104018.
4
Co-Induction of ULK-1 and AHSP mRNAs in Erythroid Precursor Cells Isolated From a Sirolimus-Treated β-Thalassemia Patient: A Case Report Study.从一名接受西罗莫司治疗的β地中海贫血患者分离的红系前体细胞中ULK-1和AHSP mRNA的共诱导：一项病例报告研究
Br J Biomed Sci. 2025 Jun 27;82:14311. doi: 10.3389/bjbs.2025.14311. eCollection 2025.
5
Precise measurement of CRISPR genome editing outcomes through single-cell DNA sequencing.通过单细胞DNA测序精确测量CRISPR基因组编辑结果。
Mol Ther Methods Clin Dev. 2025 Mar 14;33(2):101449. doi: 10.1016/j.omtm.2025.101449. eCollection 2025 Jun 12.
6
Multilayered HIV-1 resistance in HSPCs through CCR5 Knockout and B cell secretion of HIV-inhibiting antibodies.通过CCR5基因敲除和B细胞分泌HIV抑制抗体在造血干细胞中实现多层HIV-1抗性。
Nat Commun. 2025 Apr 1;16(1):3103. doi: 10.1038/s41467-025-58371-8.
7
A differentiated β-globin gene replacement strategy uses heterologous introns to restore physiological expression.一种差异化的β-珠蛋白基因替代策略利用异源内含子来恢复生理性表达。
Mol Ther. 2025 Apr 2;33(4):1407-1419. doi: 10.1016/j.ymthe.2025.02.036. Epub 2025 Feb 28.
8
MetaQ: fast, scalable and accurate metacell inference via single-cell quantization.MetaQ：通过单细胞量化实现快速、可扩展且准确的元细胞推断。
Nat Commun. 2025 Jan 31;16(1):1205. doi: 10.1038/s41467-025-56424-6.
9
Engineering synthetic signaling receptors to enable erythropoietin-free erythropoiesis.工程化合成信号受体以实现无促红细胞生成素的红细胞生成。
Nat Commun. 2025 Jan 29;16(1):1140. doi: 10.1038/s41467-025-56239-5.
10
Dual α-globin-truncated erythropoietin receptor knockin restores hemoglobin production in α-thalassemia-derived erythroid cells.双α-珠蛋白截短型促红细胞生成素受体敲入可恢复α地中海贫血来源的红系细胞中的血红蛋白生成。
Cell Rep. 2025 Jan 28;44(1):115141. doi: 10.1016/j.celrep.2024.115141. Epub 2025 Jan 3.
本文引用的文献
1
The TRACE-Seq method tracks recombination alleles and identifies clonal reconstitution dynamics of gene targeted human hematopoietic stem cells.TRACE-Seq 方法可追踪重组等位基因，并鉴定基因靶向的人类造血干细胞的克隆重建动力学。
Nat Commun. 2021 Jan 20;12(1):472. doi: 10.1038/s41467-020-20792-y.
2
Efficient gene editing of human long-term hematopoietic stem cells validated by clonal tracking.通过克隆追踪验证的人类长期造血干细胞的高效基因编辑。
Nat Biotechnol. 2020 Nov;38(11):1298-1308. doi: 10.1038/s41587-020-0551-y. Epub 2020 Jun 29.
3
CRISPR-Edited Stem Cells in a Patient with HIV and Acute Lymphocytic Leukemia.经 CRISPR 编辑的干细胞在一名 HIV 合并急性淋巴细胞白血病患者中的应用。
N Engl J Med. 2019 Sep 26;381(13):1240-1247. doi: 10.1056/NEJMoa1817426. Epub 2019 Sep 11.
4
Human genome-edited hematopoietic stem cells phenotypically correct Mucopolysaccharidosis type I.人类基因组编辑造血干细胞在表型上纠正黏多糖贮积症 I 型。
Nat Commun. 2019 Sep 6;10(1):4045. doi: 10.1038/s41467-019-11962-8.
5
In Vivo Outcome of Homology-Directed Repair at the HBB Gene in HSC Using Alternative Donor Template Delivery Methods.使用替代供体模板递送方法在造血干细胞中HBB基因进行同源定向修复的体内结果
Mol Ther Nucleic Acids. 2019 Sep 6;17:277-288. doi: 10.1016/j.omtn.2019.05.025. Epub 2019 Jun 7.
6
Highly efficient editing of the β-globin gene in patient-derived hematopoietic stem and progenitor cells to treat sickle cell disease.高效编辑患者来源的造血干细胞和祖细胞中的β-珠蛋白基因以治疗镰状细胞病。
Nucleic Acids Res. 2019 Sep 5;47(15):7955-7972. doi: 10.1093/nar/gkz475.
7
Highly Efficient and Marker-free Genome Editing of Human Pluripotent Stem Cells by CRISPR-Cas9 RNP and AAV6 Donor-Mediated Homologous Recombination.利用 CRISPR-Cas9 RNP 和 AAV6 供体介导的同源重组对人多能干细胞进行高效、无标记的基因组编辑。
Cell Stem Cell. 2019 May 2;24(5):821-828.e5. doi: 10.1016/j.stem.2019.04.001.
8
Gene correction for SCID-X1 in long-term hematopoietic stem cells.X 连锁重症联合免疫缺陷病 1 型的基因矫正：长期造血干细胞。
Nat Commun. 2019 Apr 9;10(1):1634. doi: 10.1038/s41467-019-09614-y.
9
Precise Gene Editing Preserves Hematopoietic Stem Cell Function following Transient p53-Mediated DNA Damage Response.精确的基因编辑可保留短暂的 p53 介导的 DNA 损伤反应后造血干细胞的功能。
Cell Stem Cell. 2019 Apr 4;24(4):551-565.e8. doi: 10.1016/j.stem.2019.02.019. Epub 2019 Mar 21.
10
Intrabone hematopoietic stem cell gene therapy for adult and pediatric patients affected by transfusion-dependent ß-thalassemia.经输血依赖型β-地中海贫血影响的成年和儿科患者的骨内造血干细胞基因治疗。
Nat Med. 2019 Feb;25(2):234-241. doi: 10.1038/s41591-018-0301-6. Epub 2019 Jan 21.
Zotero 插件