Allele-Specific Gene Editing Rescues Pathology in a Human Model of Charcot-Marie-Tooth Disease Type 2E.
Allele-Specific Gene Editing Rescues Pathology in a Human Model of Charcot-Marie-Tooth Disease Type 2E.
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DOI：
10.3389/fcell.2021.723023
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发表时间：
2021
影响因子：
5.5
通讯作者：
Judge LM
中科院分区：
文献类型：
--
作者：
Feliciano CM;Wu K;Watry HL;Marley CBE;Ramadoss GN;Ghanim HY;Liu AZ;Zholudeva LV;McDevitt TC;Saporta MA;Conklin BR;Judge LM
许多神经肌肉疾病是由显性错义突变引起的，导致显性阴性或功能获得性病理。这类疾病通过药物治疗或基因增强治疗具有挑战性，因为这些策略可能无法消除突变蛋白或RNA的影响。因此，这些主要疾病严重缺乏有效的治疗方法，常常导致严重残疾或死亡。通过基因编辑靶向灭活显性疾病等位基因是一种很有前途的方法，有可能通过单一治疗完全消除病理原因。在这里，我们证明了在轴突Charcot-Marie-Tooth（CMT）疾病的人类模型中等位基因特异性CRISPR基因编辑挽救了由神经丝轻链基因（NEFL，CMT 2 E型）中的显性错义突变引起的病理。我们利用一种快速有效的方法从来自CMT 2 E患者的人诱导多能干细胞（iPSC）产生脊髓运动神经元。在分化的早期时间点上，肥大的运动神经元概括了已知的病理表型，包括神经丝轻链蛋白在神经元细胞体中的异常积累。我们使用Cas9酶选择性地灭活患者iPSC中的疾病NEFL等位基因以在致病性N98 S突变处引入移码。携带这种等位基因特异性移码的运动神经元表现出与精确校正突变的同基因对照相比的疾病表型的改善。我们的研究结果验证了等位基因特异性基因编辑作为CMT 2 E的治疗方法，以及作为沉默任何基因中的显性突变的有希望的策略，其中杂合功能丧失是良好耐受的。这突出了基因编辑作为治疗目前无法治疗的显性神经系统疾病的潜力。
Many neuromuscular disorders are caused by dominant missense mutations that lead to dominant-negative or gain-of-function pathology. This category of disease is challenging to address via drug treatment or gene augmentation therapy because these strategies may not eliminate the effects of the mutant protein or RNA. Thus, effective treatments are severely lacking for these dominant diseases, which often cause severe disability or death. The targeted inactivation of dominant disease alleles by gene editing is a promising approach with the potential to completely remove the cause of pathology with a single treatment. Here, we demonstrate that allele-specific CRISPR gene editing in a human model of axonal Charcot-Marie-Tooth (CMT) disease rescues pathology caused by a dominant missense mutation in the neurofilament light chain gene (NEFL, CMT type 2E). We utilized a rapid and efficient method for generating spinal motor neurons from human induced pluripotent stem cells (iPSCs) derived from a patient with CMT2E. Diseased motor neurons recapitulated known pathologic phenotypes at early time points of differentiation, including aberrant accumulation of neurofilament light chain protein in neuronal cell bodies. We selectively inactivated the disease NEFL allele in patient iPSCs using Cas9 enzymes to introduce a frameshift at the pathogenic N98S mutation. Motor neurons carrying this allele-specific frameshift demonstrated an amelioration of the disease phenotype comparable to that seen in an isogenic control with precise correction of the mutation. Our results validate allele-specific gene editing as a therapeutic approach for CMT2E and as a promising strategy to silence dominant mutations in any gene for which heterozygous loss-of-function is well tolerated. This highlights the potential for gene editing as a therapy for currently untreatable dominant neurologic diseases.
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影响因子：
39.3
作者：
Matsa E;Dixon JE;Medway C;Georgiou O;Patel MJ;Morgan K;Kemp PJ;Staniforth A;Mellor I;Denning C
通讯作者：
Denning C
影响因子：
3.5
作者：
Abe, Akiko;Numakura, Chikahiko;Hayasaka, Kiyoshi
通讯作者：
Hayasaka, Kiyoshi
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：
9.9
作者：
Lu CH;Macdonald-Wallis C;Gray E;Pearce N;Petzold A;Norgren N;Giovannoni G;Fratta P;Sidle K;Fish M;Orrell R;Howard R;Talbot K;Greensmith L;Kuhle J;Turner MR;Malaspina A
通讯作者：
Malaspina A
影响因子：
13.6
作者：
Gaj T;Ojala DS;Ekman FK;Byrne LC;Limsirichai P;Schaffer DV
通讯作者：
Schaffer DV