A deep intronic splice–altering AIRE variant causes APECED syndrome through antisense oligonucleotide-targetable pseudoexon inclusion,Science Translational Medicine

当前位置： X-MOL 学术 › Sci. Transl. Med. › 论文详情

Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)

A deep intronic splice–altering AIRE variant causes APECED syndrome through antisense oligonucleotide-targetable pseudoexon inclusion
Science Translational Medicine ( IF 15.8 ) Pub Date : 2024-09-18 , DOI: 10.1126/scitranslmed.adk0845
Sebastian Ochoa ₁ , Amy P Hsu ₁ , Andrew J Oler ₂ , Dhaneshwar Kumar ₃ , Daniel Chauss ₃ , Jan Piet van Hamburg ₄ , Gustaaf G van Laar ₄ , Vasileios Oikonomou ₁ , Sundar Ganesan ₅ , Elise M N Ferré ₁ , Monica M Schmitt ₁ , Tom DiMaggio ₁ , Princess Barber ₁ , Gregory M Constantine ₆ , Lindsey B Rosen ₁ , Paul G Auwaerter ₇ , Bhumika Gandhi ₈ , Jennifer L Miller ₉ , Rachel Eisenberg ₁₀ , Arye Rubinstein _{10,

11} , Edith Schussler ₁₂ , Erjola Balliu ₁₃ , Vandana Shashi _{14,

15} , Olaf Neth ₁₆ , Peter Olbrich _{16,

17} , Kim My Le _{18,

19} , Nanni Mamia ₁₉ , Saila Laakso _{19,

20,

21} , Pasi I Nevalainen ₂₂ , Juha Grönholm _{18,

19} , Mikko R J Seppänen _{18,

19,

23} , Louis Boon ₂₂ , Gulbu Uzel ₁ , Luis M Franco ₂₄ , Theo Heller ₂₅ , Karen K Winer ₂₆ , Rajarshi Ghosh ₂₇ , Bryce A Seifert ₂₇ , Magdalena Walkiewicz ₂₇ , Luigi D Notarangelo ₁ , Qing Zhou ₂₈ , Ivona Askentijevich ₂₈ , William Gahl ₂₉ , Cliffton L Dalgard _{30,

31} , Lalith Perera ₃₂ , Behdad Afzali ₃ , Sander W Tas ₄ , Steven M Holland ₁ , Michail S Lionakis ₁

Affiliation

Laboratory of Clinical Immunology and Microbiology, National Institute of Allergy and Infectious Diseases (NIAID), National Institutes of Health (NIH), Bethesda, MD 20892, USA.
Bioinformatics and Computational Biosciences Branch, Office of Cyber Infrastructure and Computational Biology, NIAID, NIH, Bethesda, MD 20892, USA.
Immunoregulation Section, Kidney Diseases Branch, National Institute of Diabetes and Digestive and Kidney Diseases, NIH, Bethesda, MD 20892, USA.
Departments of Rheumatology and Clinical Immunology and Experimental Immunology, Amsterdam UMC, University of Amsterdam, Amsterdam, 1105 AZ, Netherlands.
Biological Imaging Section, Research Technologies Branch, NIAID, NIH, Bethesda, MD 20892, USA.
Laboratory of Parasitic Diseases, NIAID, NIH, Bethesda, MD 20892, USA.
Sherrilyn and Ken Fisher Center for Environmental Infectious Diseases, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
Division of Internal Medicine-Pediatrics, Department of Medicine, Medstar Georgetown University Hospital, Washington, DC 20007, USA.
Division of Pediatric Endocrinology, Department of Pediatrics, Feinberg School of Medicine, Northwestern University, Chicago, IL 60611, USA.
Division of Allergy and Immunology, Department of Pediatrics, Montefiore Medical Center, Bronx, NY 10467, USA.
Department of Microbiology and Immunology, Montefiore Medical Center, Bronx, NY 10467, USA.
Division of Pulmonary, Allergy, and Immunology, Department of Pediatrics, Weill Cornell Medicine, New York, NY 10065, USA.
Department of Endocrinology, Diabetes and Metabolism, Lakeland Regional Health Grasslands Campus, Lakeland, FL 33803, USA.
Division of Medical Genetics, Department of Pediatrics, Duke University School of Medicine, Durham, NC 27710, USA.
Undiagnosed Diseases Network, Duke University Medical Center, Durham, NC 27710, USA.
Inborn Errors of Immunity Laboratory, Biomedicine Institute in Seville (IBiS), University of Seville/CSIC, "Red de Investigación Translacional en Infectología Pediátrica," Paediatric Infectious Diseases, Rheumatology and Immunology Unit, Virgen del Rocío University Hospital, Seville 41013, Spain.
Departamento de Farmacología, Pediatría y Radiología, Facultad de Medicina, Universidad de Sevilla, Seville 41004, Spain.
Translational Immunology Research Program, University of Helsinki, Helsinki 00014, Finland.
Pediatric Research Center, New Children's Hospital, University of Helsinki and HUS Helsinki University Hospital, Helsinki 00290, Finland.
Folkhälsan Research Center, Helsinki 00250, Finland.
Research Program for Clinical and Molecular Metabolism, Faculty of Medicine, University of Helsinki, Helsinki 00014, Finland.
JJP Biologics, Warsaw 00-728, Poland.
European Reference Network Rare Immunodeficiency Autoinflammatory and Autoimmune Diseases Network (ERN RITA) Core Center, Utrecht, 3584 CX, Netherlands.
Functional Immunogenomics Unit, Systemic Autoimmunity Branch, National Institute of Arthritis and Musculoskeletal and Skin Diseases, NIH, Bethesda, MD 20892, USA.
Liver Diseases Branch, National Institute of Diabetes and Digestive and Kidney Diseases, NIH, Bethesda, MD 20892, USA.
Pediatric Growth and Nutrition Branch, National Institute of Child Health and Human Development, NIH, Bethesda, MD 20892, USA.
Centralized Sequencing Program, Division of Intramural Research, NIAID, NIH, Bethesda, MD 20892, USA.
Inflammatory Disease Section, National Human Genome Research Institute, NIH, Bethesda, MD 20892, USA.
Medical Genetics Branch, National Human Genome Research Institute, and NIH Undiagnosed Diseases Program, Common Fund, Office of the Director, NIH, Bethesda, MD 20892, USA.
Department of Anatomy, Physiology, and Genetics, Uniformed Services University of the Health Sciences, Bethesda, MD 20814, USA.
American Genome Center, Uniformed Services University of the Health Sciences, Bethesda, MD 20814, USA.
Genome Integrity and Structural Biology Laboratory, National Institute of Environmental Health Sciences, NIH, Research Triangle Park, NC 27709, USA.

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.

中文翻译：

改变深度内含子剪接的 AIRE 变体通过反义寡核苷酸靶向伪外显子包涵引起 APECED 综合征

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因子（AIRE）基因的双等位基因有害变异引起。我们前瞻性评估了 104 例临床诊断为 APECED 综合征的患者，并从 14 个亲属中确定了 17 例（16%）患者，这些患者在外显子或侧翼内含子区域缺乏双等位基因 AIRE 变异;15 人有波多黎各血统。通过全基因组测序，我们在所有 17 例患者中鉴定了一个与疾病共分离的深度内含子 AIRE 变体（c.1504-818 G>A）。我们开发了一种表达 AIRE 的原代患者单核细胞衍生的树突细胞培养系统，并证明 c.1504-818 G>A 会产生一个隐蔽的剪接位点并激活 109 个碱基对移码伪外显子的包含。我们还在患者来源的淋巴母细胞样细胞系（LCL）中发现了低水平 AIRE 表达，并证实了在独立的胸腺外表达 AIRE 的细胞系中包含假外显子。通过对 AIRE 转染的人胚胎肾 293 和胸腺上皮细胞 4D6 细胞的蛋白质建模和转录组学分析，我们发现这种变体改变了 AIRE 蛋白的羧基末端，消除了其功能。最后，我们开发了一种反义寡核苷酸（ASO），可逆转假外显子包涵并恢复 LCL 中的正常 AIRE 转录序列。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 AIRE 变体的创始 APECED 变体，并揭示了假外显子包涵是 APECED 背后的 ASO 可逆遗传机制。

更新日期：2024-09-18

点击分享查看原文

点击收藏

阅读更多本刊新发论文本刊介绍/投稿指南