Targeted exon skipping of a CEP290 mutation rescues Joubert syndrome phenotypes in vitro and in a murine model.
Targeted exon skipping of a CEP290 mutation rescues Joubert syndrome phenotypes in vitro and in a murine model.
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DOI：
10.1073/pnas.1809432115
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发表时间：
2018-12-04
影响因子：
11.1
通讯作者：
Sayer JA
中科院分区：
文献类型：
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作者：
Ramsbottom SA;Molinari E;Srivastava S;Silberman F;Henry C;Alkanderi S;Devlin LA;White K;Steel DH;Saunier S;Miles CG;Sayer JA
遗传性肾病的治疗具有挑战性，因为这需要纠正潜在的基因缺陷和提供治疗。在这里，我们表明，通过使用反义寡核苷酸，我们可以诱导外显子跳跃的突变外显子CEP290，来自一个纤毛病变综合征患者称为Joubert综合征的肾上皮细胞内。这种治疗将截短的CEP290蛋白拯救为接近全长的蛋白质并恢复纤毛表型。在Joubert综合征的Cep 290小鼠模型中，通过系统性治疗反义寡核苷酸可以实现外显子跳读，从而挽救睫状体和肾脏疾病表型。这项工作为遗传性肾病患者的个性化基因治疗铺平了道路。导致囊性肾病的肾纤毛病变的基因治疗将为目前的治疗提供真实的进步。CEP290的突变是一种称为Joubert综合征（JBTS）的纤毛病的基础。人类疾病表型包括脑、视网膜和肾脏疾病，其通常在生命的前二十年内进展为终末期肾衰竭（ESRF）。虽然目前无法治愈，但在诊断和ESRF之间通常有一段时间，这为治疗干预提供了一个潜在的窗口。通过研究患者活检、患者来源的肾细胞和小鼠模型，我们确定了初级纤毛异常伸长是CEP290相关JBTS的关键病理生理学特征，并表明反义寡核苷酸（阿索）诱导的突变外显子（41，G1890*）剪接恢复了患者细胞中的蛋白质表达。我们证明，ASO诱导的剪接导致外显子跳跃是耐受的，导致正确的定位CEP290蛋白的纤毛过渡区，并恢复正常的纤毛长度在患者的肾细胞。使用JBTS的基因陷阱Cep290小鼠模型，我们表明全身性阿索治疗可以减少体内患病肾脏的囊性负担。这些发现表明，阿索治疗可能是一种有希望的治疗方法，用于CEP290相关性纤毛病变综合征的肾脏疾病。
The treatment of genetic kidney disease is challenging, as this requires both the correction of the underlying gene defect and the delivery of the treatment. Here we show that by using antisense oligonucleotides, we can induce exon skipping of a mutated exon in CEP290, within renal epithelial cells derived from a patient with a ciliopathy syndrome called Joubert syndrome. This treatment rescues the truncated CEP290 protein to a near full-length protein and restores the ciliary phenotype. In a Cep290 murine model of Joubert syndrome, exon skipping is achievable with systemic treatment of an antisense oligonucleotide, which rescues both the ciliary and kidney disease phenotypes. This work paves the way toward personalized genetic therapies in patients with inherited kidney diseases. Genetic treatments of renal ciliopathies leading to cystic kidney disease would provide a real advance in current therapies. Mutations in CEP290 underlie a ciliopathy called Joubert syndrome (JBTS). Human disease phenotypes include cerebral, retinal, and renal disease, which typically progresses to end stage renal failure (ESRF) within the first two decades of life. While currently incurable, there is often a period of years between diagnosis and ESRF that provides a potential window for therapeutic intervention. By studying patient biopsies, patient-derived kidney cells, and a mouse model, we identify abnormal elongation of primary cilia as a key pathophysiological feature of CEP290-associated JBTS and show that antisense oligonucleotide (ASO)-induced splicing of the mutated exon (41, G1890*) restores protein expression in patient cells. We demonstrate that ASO-induced splicing leading to exon skipping is tolerated, resulting in correct localization of CEP290 protein to the ciliary transition zone, and restoration of normal cilia length in patient kidney cells. Using a gene trap Cep290 mouse model of JBTS, we show that systemic ASO treatment can reduce the cystic burden of diseased kidneys in vivo. These findings indicate that ASO treatment may represent a promising therapeutic approach for kidney disease in CEP290-associated ciliopathy syndromes.
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影响因子：
12.3
作者：
Sanders AA;de Vrieze E;Alazami AM;Alzahrani F;Malarkey EB;Sorusch N;Tebbe L;Kuhns S;van Dam TJ;Alhashem A;Tabarki B;Lu Q;Lambacher NJ;Kennedy JE;Bowie RV;Hetterschijt L;van Beersum S;van Reeuwijk J;Boldt K;Kremer H;Kesterson RA;Monies D;Abouelhoda M;Roepman R;Huynen MH;Ueffing M;Russell RB;Wolfrum U;Yoder BK;van Wijk E;Alkuraya FS;Blacque OE
通讯作者：
Blacque OE
影响因子：
4.4
作者：
Roshon, M;DeGregori, JV;Ruley, HE
通讯作者：
Ruley, HE
影响因子：
13.6
作者：
Smith, Laurie A.;Bukanov, Nikolay O.;Ibraghimov-Beskrovnaya, Oxana
通讯作者：
Ibraghimov-Beskrovnaya, Oxana
影响因子：
4.4
作者：
Littink, Karin W.;Pott, Jan-Willem R.;den Hollander, Anneke I.
通讯作者：
den Hollander, Anneke I.
影响因子：
21.3
作者：
通讯作者：
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