PDF(Pubmed)
Abstract：
Precision medicine is rapidly gaining recognition in the field of (ultra)rare conditions, where only a few individuals in the world are affected. Clinical trial design for a small number of patients is extremely challenging, and for this reason, the development of N-of-1 strategies is explored to accelerate customized therapy design for rare cases. A strong candidate for this approach is Stargardt disease (STGD1), an autosomal recessive macular degeneration characterized by high genetic and phenotypic heterogeneity. STGD1 is caused by pathogenic variants in ABCA4, and amongst them, several deep-intronic variants alter the pre-mRNA splicing process, generally resulting in the insertion of pseudoexons (PEs) into the final transcript. In this study, we describe a 10-year-old girl harboring the unique deep-intronic ABCA4 variant c.6817-713A>G. Clinically, she presents with typical early-onset STGD1 with a high disease symmetry between her two eyes. Molecularly, we designed antisense oligonucleotides (AONs) to block the produced PE insertion. Splicing rescue was assessed in three different in vitro models: HEK293T cells, fibroblasts, and photoreceptor precursor cells, the last two being derived from the patient. Overall, our research is intended to serve as the basis for a personalized N-of-1 AON-based treatment to stop early vision loss in this patient.
摘要：
精准医学在（超）稀有条件领域迅速获得认可，世界上只有少数人受到影响。针对少数患者的临床试验设计极具挑战性,出于这个原因，探讨了N-of-1策略的开发,以加速针对罕见病例的定制治疗设计.这种方法的一个强有力的候选者是Stargardt病（STGD1），常染色体隐性黄斑变性，具有高度遗传和表型异质性。STGD1是由ABCA4的致病变异引起的，其中，几种深内含子变体改变了前mRNA剪接过程，通常导致假外显子(PE)插入到最终的转录物中。在这项研究中,我们描述了一个10岁的女孩，她拥有独特的深内含子ABCA4变体c.6817-713A>G。临床上,她表现出典型的早发性STGD1，两只眼睛之间具有高度的疾病对称性。分子上，我们设计了反义寡核苷酸(AON)来阻断产生的PE插入。在三种不同的体外模型中评估剪接拯救：HEK293T细胞，成纤维细胞,和感光前体细胞，最后两个来自病人。总的来说,我们的研究旨在作为基于N-of-1AON的个性化治疗的基础,以阻止该患者的早期视力丧失.
