A proactive missense variant atlas for the Autoimmune Regulator

项目摘要/摘要 自身免疫调节因子(AIRE)基因的致病变异导致原发免疫缺乏症 自身免疫性多内分泌综合征1型(APS-1)；也称为自身免疫性多内分泌病 念珠菌病和外胚层营养不良)，表现为包括慢性黏膜皮肤的症状 念珠菌病、甲状旁腺功能减退症和爱迪生病以及其他自身免疫症状。AIRE防止 通过在发育中的T细胞的利基中增加组织特异性基因的表达来实现自身免疫，从而 用于在T细胞成熟之前消除新出现的自我反应性T细胞。AIRE的排序是一种 诊断APS-1的工具越来越普遍，早期诊断可以使患者受益。然而，诊断是 受限于这样一个事实，即三分之二的变体是错义的，而超过一半的错义变体 被发现的是“意义未知的”。在培养细胞中测试变异体的功能可以提供强大的 更有信息量的变异分类的证据，但这些测试是反应性的，通常是几个月或几个月 在患者身上首次观察到新的变异数年后。这个问题的一个积极的解决方案是高效地 评估所有可能的错义变体的功能，以生成序列功能图。这样的地图可以 提供即时和准确的证据来诊断疾病，即使是以前从未见过的变种。我们有 开发并验证了一种可扩展的测试AIRE错义变体的方法，使用具有 胰岛素启动子活性的荧光报告，并正在探索其他可扩展的检测方法。我们会： 1)生成序列-功能图，以实现对AIRE错义的更准确的临床解释 变种和； 2)验证和刻画AIRE序列-功能图。 AIRE的序列-功能图谱将为快速和明确的基因诊断提供关键的主动证据 APS-1，并最终获得更积极的患者结果。

Project Summary/Abstract Pathogenic variants of the Autoimmune Regulator (AIRE) gene cause the primary immunodeficiency disease Autoimmune Polyendocrine Syndrome Type 1 (APS-1; also called Autoimmune Polyendocrinopathy with Candidiasis and Ectodermal Dystrophy), which manifests as symptoms including chronic mucocutaneous candidiasis, hypoparathyroidism, and Addison’s disease among other autoimmune symptoms. AIRE prevents autoimmunity by increasing the expression of tissue-specific genes in the niche of developing T cells, allowing for the elimination of emerging self-reactive T-cells before they can mature. Sequencing of AIRE is an increasingly common tool to diagnose APS-1, and earlier diagnosis can benefit patients. However, diagnosis is limited by the fact that two thirds of variants are missense, and that more than half of the missense variants found are “of unknown significance”. Testing the function of variants in cultured cells can provide strong evidence for more informative variant classification, but these tests are carried out reactively, often months or years after a new variant is first observed in a patient. A proactive solution to this problem is to efficiently assess the function of all possible missense variants to generate a sequence-function map. Such maps can provide immediate and accurate evidence to diagnose disease, even for never-before-seen variants. We have developed and validated one scalable assay for testing AIRE missense variants, using human cells with a fluorescent reporter of insulin promoter activity, and are exploring other scalable assays. We will: 1) Generate a sequence-function map to enable more accurate clinical interpretation of AIRE missense variants and; 2) Validate and characterize the AIRE sequence-function map. A sequence-function map of AIRE will provide key proactive evidence for rapid and definitive genetic diagnosis of APS-1, and ultimately more positive patient outcomes.