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Brief Report: Novel UNC13D Intronic Variant Disrupting an NF‐κB Enhancer in a Patient With Recurrent Macrophage Activation Syndrome and Systemic Juvenile Idiopathic Arthritis

Title: Brief Report: Novel UNC13D Intronic Variant Disrupting an NF‐κB Enhancer in a Patient With Recurrent Macrophage Activation Syndrome and Systemic Juvenile Idiopathic Arthritis
Authors: Schulert, Grant S.; Zhang, Mingce; Husami, Ammar; Fall, Ndate; Brunner, Hermine; Zhang, Kejian; Cron, Randy Q.; Grom, Alexei A.
Contributors: National Institutes of Health; Rheumatology Research Foundation; Cincinnati Children’s Research Foundation; Histiocytosis Association
Source: Arthritis & Rheumatology ; volume 70, issue 6, page 963-970 ; ISSN 2326-5191 2326-5205
Publisher Information: Wiley
Publication Year: 2018
Collection: Wiley Online Library (Open Access Articles via Crossref)
Description: Objective Macrophage activation syndrome (MAS) is a life‐threatening complication of systemic juvenile idiopathic arthritis (JIA) and has pathologic similarity to hemophagocytic lymphohistiocytosis (HLH). Intronic variants in UNC13D are found in patients with familial HLH type 3 (FHLH3), but the role of noncoding variants in MAS is unknown. The objective of this study was to identify deep intronic UNC13D variants in patients with MAS. Methods A custom enrichment library was constructed to sequence a genomic region of ~1 Mb flanking UNC13D in 24 patients with systemic JIA, recurrent MAS, and negative results of prior genetic (exon/coding) testing. The functional consequences of intronic variants were assessed using quantitative polymerase chain reaction in patient‐derived peripheral blood mononuclear cells (PBMCs), electromobility shift assay, in vitro transcriptional enhancer assays, and natural killer (NK) cell degranulation assays. Results We evaluated a patient with systemic JIA and recurrent MAS in whom a novel functional intronic variant in UNC13D , c.117+143A>G, was observed. This variant occurred in a proposed regulatory region that drives lymphocyte‐specific UNC13D expression and is associated with reduced transcript levels in patient PBMCs. This variant also disrupted NF‐κB 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 noncoding variant hit rate of 8.3% (2 of 24). Conclusion These findings highlight the notion that intronic variants in key regulatory regions may be associated with MAS in patients with systemic JIA and support deep sequencing approaches when causative coding variants are not identified.
Document Type: article in journal/newspaper
Language: English
DOI: 10.1002/art.40438
Availability: https://doi.org/10.1002/art.40438; https://api.wiley.com/onlinelibrary/tdm/v1/articles/10.1002%2Fart.40438; https://onlinelibrary.wiley.com/doi/full/10.1002/art.40438
Rights: http://onlinelibrary.wiley.com/termsAndConditions#vor
Accession Number: edsbas.B82A48F4
Database: BASE