In vivo base editing rescues Hutchinson-Gilford progeria syndrome in mice.
In vivo base editing rescues Hutchinson-Gilford progeria syndrome in mice.
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DOI：
10.1038/s41586-020-03086-7
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发表时间：
2021-01
期刊：
影响因子：
64.8
通讯作者：
Liu DR
中科院分区：
文献类型：
--
作者：
Koblan LW;Erdos MR;Wilson C;Cabral WA;Levy JM;Xiong ZM;Tavarez UL;Davison LM;Gete YG;Mao X;Newby GA;Doherty SP;Narisu N;Sheng Q;Krilow C;Lin CY;Gordon LB;Cao K;Collins FS;Brown JD;Liu DR
Hutchinson-Gilford早老综合征（HGPS）是由核纤层蛋白A基因LMNA的显性负性C·G → T·A突变（c.1824 C>T，G608 G）引起的。这种突变导致RNA错误剪接，产生早老蛋白，这是一种毒性蛋白，可诱导快速衰老并将寿命缩短至约14年。腺嘌呤碱基编辑器（ABE）执行靶向A·T到G·C碱基对转换，副产物最少，不需要双链DNA断裂或供体DNA模板。在这里，我们描述了使用ABE直接纠正培养的早衰症患者来源的成纤维细胞和HGPS小鼠模型中的致病性HGPS突变。ABE向患者来源的成纤维细胞的慢病毒递送导致致病等位基因的~90%校正、RNA错误剪接的减轻、早老蛋白水平的降低以及核异常的校正。无偏脱靶DNA和RNA分析未检测到经处理的患者来源的成纤维细胞中的脱靶编辑活性。在人类LMNA c.1824 C>T等位基因纯合的转基因小鼠中，单次眶后注射编码ABE的腺相关病毒9（AAV 9）导致致病性突变的实质性持久校正（注射后6个月，在各种器官中约20-60%）、正常RNA剪接的恢复和早老蛋白的减少。体内碱基编辑挽救了血管病理学，保留了血管平滑肌细胞计数并防止了外膜纤维化。在P14时单次注射ABE AAV 9改善了动物活力，并将中位寿命从215天大大延长至510天。这些发现支持了体内碱基编辑通过直接纠正疾病的根本原因来治疗HGPS和其他遗传疾病的潜力。
Hutchinson-Gilford progeria syndrome (HGPS) is typically caused by a dominant-negative C•G-to-T•A mutation (c.1824 C>T, G608G) in LMNA, the nuclear lamin A gene. This mutation causes RNA mis-splicing that produces progerin, a toxic protein that induces rapid aging and shortens lifespan to ~14 years. Adenine base editors (ABEs) perform targeted A•T-to-G•C base pair conversion with minimal byproducts and without requiring double-strand DNA breaks or donor DNA templates. Here, we describe the use of an ABE to directly correct the pathogenic HGPS mutation in cultured progeria patient-derived fibroblasts and in a mouse model of HGPS. Lentiviral delivery of ABE to patient-derived fibroblasts results in ~90% correction of the pathogenic allele, mitigation of RNA mis-splicing, reduced progerin levels, and correction of nuclear abnormalities. Unbiased off-target DNA and RNA analysis did not detect off-target editing activity in treated patient-derived fibroblasts. In transgenic mice homozygous for the human LMNA c.1824 C>T allele, a single retro-orbital injection of adeno-associated virus 9 (AAV9) encoding the ABE resulted in substantial, durable correction of the pathogenic mutation (~20-60% across various organs 6 months post-injection), restoration of normal RNA splicing, and reduction of progerin protein. In vivo base editing rescued vascular pathology, preserving vascular smooth muscle cell counts and preventing adventitial fibrosis. A single ABE AAV9 injection at P14 improved animal vitality and greatly extended median lifespan from 215 to 510 days. These findings support the potential of in vivo base editing to treat HGPS, and other genetic diseases, by directly correcting the root cause of disease.
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影响因子：
46.9
作者：
Doench JG;Fusi N;Sullender M;Hegde M;Vaimberg EW;Donovan KF;Smith I;Tothova Z;Wilen C;Orchard R;Virgin HW;Listgarten J;Root DE
通讯作者：
Root DE
影响因子：
82.9
作者：
Beyret, Ergin;Liao, Hsin-Kai;Belmonte, Juan Carlos Izpisua
通讯作者：
Belmonte, Juan Carlos Izpisua
影响因子：
17.1
作者：
Cao, Kan;Graziotto, John J.;Collins, Francis S.
通讯作者：
Collins, Francis S.
使用下一代 DNA 测序数据进行变异发现和基因分型的框架。
影响因子：
30.8
作者：
通讯作者：
--
影响因子：
37.8
作者：
Gordon LB;Massaro J;D'Agostino RB Sr;Campbell SE;Brazier J;Brown WT;Kleinman ME;Kieran MW;Progeria Clinical Trials Collaborative
通讯作者：
Progeria Clinical Trials Collaborative