深内含子剪接改变的AIRE变体通过反义寡核苷酸靶向伪外显子包涵导致APECED综合征

IF 15.8 1区 医学 Q1 CELL BIOLOGY Science Translational Medicine Pub Date : 2024-09-18 DOI:10.1126/scitranslmed.adk0845
Sebastian Ochoa, Amy P. Hsu, Andrew J. Oler, Dhaneshwar Kumar, Daniel Chauss, Jan Piet van Hamburg, Gustaaf G. van Laar, Vasileios Oikonomou, Sundar Ganesan, Elise M. N. Ferré, Monica M. Schmitt, Tom DiMaggio, Princess Barber, Gregory M. Constantine, Lindsey B. Rosen, Paul G. Auwaerter, Bhumika Gandhi, Jennifer L. Miller, Rachel Eisenberg, Arye Rubinstein, Edith Schussler, Erjola Balliu, Vandana Shashi, Olaf Neth, Peter Olbrich, Kim My Le, Nanni Mamia, Saila Laakso, Pasi I. Nevalainen, Juha Grönholm, Mikko R. J. Seppänen, Louis Boon, Gulbu Uzel, Luis M. Franco, Theo Heller, Karen K. Winer, Rajarshi Ghosh, Bryce A. Seifert, Magdalena Walkiewicz, Luigi D. Notarangelo, Qing Zhou, Ivona Askentijevich, William Gahl, Cliffton L. Dalgard, Lalith Perera, Behdad Afzali, Sander W. Tas, Steven M. Holland, Michail S. Lionakis
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引用次数: 0

摘要

自身免疫性多内分泌病-念珠菌病-外胚层营养不良症(APECED)是一种危及生命的单基因自身免疫性疾病,主要由自身免疫调节剂(AIRE)基因中的双倍性有害变体引起。我们对 104 名临床诊断为 APECED 综合征的患者进行了前瞻性评估,并从 14 个在外显子或侧翼内含子区缺乏双倍性 AIRE 变异的家族中发现了 17 名患者(16%);其中 15 名患者有波多黎各血统。通过全基因组测序,我们在所有 17 名患者中发现了一个与疾病共存的 AIRE 深度内含子变异(c.1504-818 G>A)。我们建立了一个表达 AIRE 的原发性患者单核细胞衍生树突细胞培养系统,并证明 c.1504-818 G>A 创建了一个隐性剪接位点,并激活了 109 碱基对移帧假外显子的包含。我们还在患者衍生的淋巴母细胞系(LCLs)中发现了低水平的 AIRE 表达,并在独立的外胚层 AIRE 表达细胞系中证实了假外显子的包含。通过对转染 AIRE 的人胚肾 293 细胞和胸腺上皮细胞 4D6 细胞进行蛋白质建模和转录组分析,我们发现该变体改变了 AIRE 蛋白的羧基端,从而削弱了其功能。最后,我们开发了一种反义寡核苷酸(ASO),它能逆转假外显子的包含,并恢复 LCLs 中 AIRE 转录本的正常序列。因此,我们的研究结果显示,c.1504-818 G>A 是波多黎各人群中导致 APECED 的 AIRE 基因变异,并揭示了假外显子包涵是一种 ASO 可逆的 APECED 遗传机制。
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A deep intronic splice–altering AIRE variant causes APECED syndrome through antisense oligonucleotide-targetable pseudoexon inclusion
Autoimmune polyendocrinopathy-candidiasis-ectodermal dystrophy (APECED) is a life-threatening monogenic autoimmune disorder primarily caused by biallelic deleterious variants in the autoimmune regulator (AIRE) gene. We prospectively evaluated 104 patients with clinically diagnosed APECED syndrome and identified 17 patients (16%) from 14 kindreds lacking biallelic AIRE variants in exons or flanking intronic regions; 15 had Puerto Rican ancestry. Through whole-genome sequencing, we identified a deep intronic AIRE variant (c.1504-818 G>A) cosegregating with the disease in all 17 patients. We developed a culture system of AIRE-expressing primary patient monocyte-derived dendric cells and demonstrated that c.1504-818 G>A creates a cryptic splice site and activates inclusion of a 109–base pair frame-shifting pseudoexon. We also found low-level AIRE expression in patient-derived lymphoblastoid cell lines (LCLs) and confirmed pseudoexon inclusion in independent extrathymic AIRE–expressing cell lines. Through protein modeling and transcriptomic analyses of AIRE-transfected human embryonic kidney 293 and thymic epithelial cell 4D6 cells, we showed that this variant alters the carboxyl terminus of the AIRE protein, abrogating its function. Last, we developed an antisense oligonucleotide (ASO) that reversed pseudoexon inclusion and restored the normal AIRE transcript sequence in LCLs. Thus, our findings revealed c.1504-818 G>A as a founder APECED-causing AIRE variant in the Puerto Rican population and uncovered pseudoexon inclusion as an ASO-reversible genetic mechanism underlying APECED.
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来源期刊
Science Translational Medicine
Science Translational Medicine CELL BIOLOGY-MEDICINE, RESEARCH & EXPERIMENTAL
CiteScore
26.70
自引率
1.20%
发文量
309
审稿时长
1.7 months
期刊介绍: Science Translational Medicine is an online journal that focuses on publishing research at the intersection of science, engineering, and medicine. The goal of the journal is to promote human health by providing a platform for researchers from various disciplines to communicate their latest advancements in biomedical, translational, and clinical research. The journal aims to address the slow translation of scientific knowledge into effective treatments and health measures. It publishes articles that fill the knowledge gaps between preclinical research and medical applications, with a focus on accelerating the translation of knowledge into new ways of preventing, diagnosing, and treating human diseases. The scope of Science Translational Medicine includes various areas such as cardiovascular disease, immunology/vaccines, metabolism/diabetes/obesity, neuroscience/neurology/psychiatry, cancer, infectious diseases, policy, behavior, bioengineering, chemical genomics/drug discovery, imaging, applied physical sciences, medical nanotechnology, drug delivery, biomarkers, gene therapy/regenerative medicine, toxicology and pharmacokinetics, data mining, cell culture, animal and human studies, medical informatics, and other interdisciplinary approaches to medicine. The target audience of the journal includes researchers and management in academia, government, and the biotechnology and pharmaceutical industries. It is also relevant to physician scientists, regulators, policy makers, investors, business developers, and funding agencies.
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