Brief Report: Novel UNC13D Intronic Variant Disrupting an NF-κB Enhancer in a Patient With Recurrent Macrophage Activation Syndrome and Systemic Juvenile Idiopathic Arthritis.
Brief Report: Novel UNC13D Intronic Variant Disrupting an NF-κB Enhancer in a Patient With Recurrent Macrophage Activation Syndrome and Systemic Juvenile Idiopathic Arthritis.
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DOI：
10.1002/art.40438
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发表时间：
2018-06
期刊：
影响因子：
--
通讯作者：
Grom AA
中科院分区：
文献类型：
--
作者：
Schulert GS;Zhang M;Husami A;Fall N;Brunner H;Zhang K;Cron RQ;Grom AA
巨噬细胞激活综合征(MAS)是系统性幼年特发性关节炎(SJIA)的一种危及生命的并发症，在病理上与吞噬血细胞淋巴组织细胞增多症(HLH)有相似之处。家族性人促黄体生成素-3(FHL3)患者中发现了UNC13D内含子变异，但非编码变异在MAS中的作用尚不清楚。这项研究的目的是确定MAS患者中的深内含子UNC13D变异体。构建了一个定制的富集库，对24例SJIA、复发的MAS和阴性的既往遗传(外显子/编码)检测阴性的患者进行了大约1MB的UNC13D侧翼序列测定。外周血单个核细胞(PBMC)定量聚合酶链式反应、凝胶迁移率改变分析(EMSA)、体外转录增强子分析和NK细胞脱颗粒实验评估内含子变异的功能后果。我们报告了一位患有SJIA和复发性MAS的患者，发现有一种新的功能型内含子变异体，C 117+143A>；G，该变异体发生在一个建议的调节区，该区域驱动淋巴细胞特异性的UN13D表达，并与患者PBMC中转录水平的降低相关。患者的变异体还破坏了NFκB与功能转录增强子的结合，导致体外增强子活性降低。部分下调UNC13D的表达也会导致NK细胞脱颗粒功能受损。另一名患者被确认为先前描述的UNC13D内含子变体，总的非编码变体命中率为8.3%(2/24)。这些发现强调了关键调控区域的内含子变异可能与SJIA患者的MAS相关，并支持当致病编码变异未被识别时的深度测序方法。
Macrophage activation syndrome (MAS) is a life-threatening complication of systemic juvenile idiopathic arthritis (SJIA), and has pathologic similarity to hemophagocytic lymphohistiocytosis (HLH). Intronic variants in UNC13D are found in patients with familial HLH-3 (FHL3), but the role of non-coding variants in MAS is unknown. The objective of this study was to identify deep intronic UNC13D variants in patients with MAS. A custom enrichment library was constructed to sequence approximately 1 MB flanking UNC13D in 24 patients with SJIA, recurrent MAS, and negative prior genetic (exon/coding) testing. Functional consequences of intronic variants were assessed using quantitative PCR on patient derived peripheral blood mononuclear cells (PBMC), electrophoretic mobility shift assay (EMSA), in vitro transcriptional enhancer assays, and NK cell degranulation assays. We report a patient with SJIA and recurrent MAS, found to have a novel functional intronic variant in UNC13D, c.117+143A>G. This variant occurs in a proposed regulatory region that drives lymphocyte specific UNC13D expression, and is associated with reduced transcript levels in patient PBMC. The patient’s variant also disrupts NFκB binding to a functional transcriptional enhancer, leading to reduced enhancer activity in vitro. Partial knockdown of UNC13D expression also led to impaired NK cell degranulation. An additional patient was identified with a previously described UNC13D intronic variant, for a total non-coding variant hit rate of 8.3% (2/24). These findings highlight that intronic variants in key regulatory regions may be associated with MAS in patients with SJIA, and support deep sequencing approaches when causative coding variants are not identified.
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影响因子：
20.3
作者：
Meeths, Marie;Chiang, Samuel C. C.;Bryceson, Yenan T.
通讯作者：
Bryceson, Yenan T.
影响因子：
5.5
作者：
Vastert, Sebastiaan J.;van Wijk, Richard;Kuis, Wietse
通讯作者：
Kuis, Wietse
DOI：
10.4049/jimmunol.1501284
发表时间：
2016-03-15
期刊：
Journal of immunology (Baltimore, Md. : 1950)
影响因子：
--
作者：
Zhang M;Bracaglia C;Prencipe G;Bemrich-Stolz CJ;Beukelman T;Dimmitt RA;Chatham WW;Zhang K;Li H;Walter MR;De Benedetti F;Grom AA;Cron RQ
通讯作者：
Cron RQ
影响因子：
13.3
作者：
Ravelli, Angelo;Minoia, Francesca;Cron, Randy Q.
通讯作者：
Cron, Randy Q.
DOI：
10.4049/jimmunol.1103129
发表时间：
2012-09-15
期刊：
Journal of immunology (Baltimore, Md. : 1950)
影响因子：
--
作者：
Zhang M;Clausell A;Robinson T;Yin J;Chen E;Johnson L;Weiss G;Sabbaj S;Lowe RM;Wagner FH;Goepfert PA;Kutsch O;Cron RQ
通讯作者：
Cron RQ