美国斯坦福大学Sergiu P. Pa?ca研究组开发出蒂莫西综合征的反义寡核苷酸治疗方法。该项研究成果于2024年4月24日在线发表在《自然》杂志上。

研究人员表示，蒂莫西综合征（Timothy Syndrome，TS）是一种严重的多系统疾病，以自闭症、癫痫、长QT综合征和其他神经精神疾病为特征。TS 1型（TS1）是由CACNA1C第8A号外显子（与其对应的第8号外显子相反）的可变剪接和发育丰富的功能增益变异引起的。研究人员之前在TS1患者的神经元中发现了几种表型，包括通道失活延迟、去极化诱导的钙离子上升时间延长、中间神经元迁移受损、活动依赖性树突回缩以及外显子8A意外的持续表达。研究人员推断，将CACNA1C外显子利用率从8A转换为8将是一种潜在的治疗策略。

研究人员开发了反义寡核苷酸（ASO），以有效减少体外和移植后体内人体细胞中8A外显子的包含。研究人员发现，由反义寡核苷酸介导的外显子 8A 到 8 的转换能有力地，挽救患者皮质器官组织的缺陷和前脑组装体的迁移。利用之前开发的移植平台，研究人员发现单次鞘内注射ASO可挽救患者神经元的钙变化和体内树突回缩，这表明抑制CACNA1C 8A外显子的表达是治疗TS1的一种潜在方法。总的来说，这些实验说明了基于体内和体外干细胞模型的多层次方法，如何确定逆转疾病相关神经病理生理学的策略。

附：英文原文

Title: Antisense oligonucleotide therapeutic approach for Timothy syndrome

Author: Chen, Xiaoyu, Birey, Fikri, Li, Min-Yin, Revah, Omer, Levy, Rebecca, Thete, Mayuri Vijay, Reis, Noah, Kaganovsky, Konstantin, Onesto, Massimo, Sakai, Noriaki, Hudacova, Zuzana, Hao, Jin, Meng, Xiangling, Nishino, Seiji, Huguenard, John, Paca, Sergiu P.

Issue&Volume: 2024-04-24

Abstract: Timothy syndrome (TS) is a severe, multisystem disorder characterized by autism, epilepsy, long-QT syndrome and other neuropsychiatric conditions1. TS type1 (TS1) is caused by a gain-of-function variant in the alternatively spliced and developmentally enriched CACNA1C exon8A, as opposed to its counterpart exon8. We previously uncovered several phenotypes in neurons derived from patients with TS1, including delayed channel inactivation, prolonged depolarization-induced calcium rise, impaired interneuron migration, activity-dependent dendrite retraction and an unanticipated persistent expression of exon8A2,3,4,5,6. We reasoned that switching CACNA1C exon utilization from 8A to 8 would represent a potential therapeutic strategy. Here we developed antisense oligonucleotides (ASOs) to effectively decrease the inclusion of exon8A in human cells both in vitro and, following transplantation, in vivo. We discovered that the ASO-mediated switch from exon8A to 8 robustly rescued defects in patient-derived cortical organoids and migration in forebrain assembloids. Leveraging a transplantation platform previously developed7, we found that a single intrathecal ASO administration rescued calcium changes and in vivo dendrite retraction of patient neurons, suggesting that suppression of CACNA1C exon8A expression is a potential treatment for TS1. Broadly, these experiments illustrate how a multilevel, in vivo and in vitro stem cell model-based approach can identify strategies to reverse disease-relevant neural pathophysiology.

DOI: 10.1038/s41586-024-07310-6

Source: https://www.nature.com/articles/s41586-024-07310-6