Identification of Germline Non-coding Deletions in XIAP Gene Causing XIAP Deficiency Reveals a Key Promoter Sequence
Identification of Germline Non-coding Deletions in XIAP Gene Causing XIAP Deficiency Reveals a Key Promoter Sequence
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鉴定 XIAP 基因中导致 XIAP 缺陷的种系非编码缺失，揭示关键启动子序列
DOI：
10.1007/s10875-021-01188-z
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发表时间：
2022
影响因子：
9.1
通讯作者：
Latour Sylvain
中科院分区：
文献类型：
--
作者：
Sbihi Zineb;Tanita Kay;Bachelet Camille;Bole Christine;Jabot-Hanin Fabienne;…Isoda Takeshi（24人中20番目）;…;Kanegane Hirokazu;Latour Sylvain
目的：细胞凋亡蛋白连锁抑制剂（XIAP）缺乏症，也称为2型x连锁淋巴组织增生综合征（XLP-2），是一种罕见的免疫缺陷，以复发性噬血细胞性淋巴组织细胞增多症、脾肿大和炎症性肠病为特征。据报道，包括错义、无义、移码和编码外显子缺失在内的XIAP变异会导致XIAP缺陷。我们研究了三名具有xlp -2样临床特征的免疫缺陷男童。通过全外显子组测序（WES）未发现XIAP编码外显子的遗传变异，尽管患者表现出XIAP表达完全缺失。方法对3例患者的DNA样本进行全基因定位测序（targeted next-generation sequencing， NGS）。通过HEK细胞中的基因报告基因表达测定和原代T细胞中的CRISPR-Cas9基因组编辑来评估分子研究。结果xiapsngs在患者中鉴定出三种不同的非编码缺失，预测这些缺失是由重复DNA序列驱动的。这些缺失共享一个839bp的共同区域，该区域包含xiap1的第一个非编码外显子，并且包含活性启动子特异性的调控元件和标记。此外，我们发现在839bp中，外显子具有转录活性。最后，CRISPR-Cas9在原代细胞中删除外显子降低了XIAP蛋白的表达。这些结果确定了一个关键的启动子序列包含在xiap的第一个非编码外显子中。重要的是，本研究强调，当编码外显子未发现变异时，应在遗传诊断中考虑目前未被WES捕获的非编码外显子的测序。
PurposeX-linked inhibitor of apoptosis protein (XIAP) deficiency, also known as the X-linked lymphoproliferative syndrome of type 2 (XLP-2), is a rare immunodeficiency characterized by recurrent hemophagocytic lymphohistiocytosis, splenomegaly, and inflammatory bowel disease. Variants inXIAPincluding missense, non-sense, frameshift, and deletions of coding exons have been reported to cause XIAP deficiency. We studied three young boys with immunodeficiency displaying XLP-2-like clinical features. No genetic variation in the coding exons ofXIAPwas identified by whole-exome sequencing (WES), although the patients exhibited a complete loss of XIAP expression.MethodsTargeted next-generation sequencing (NGS) of the entire locus ofXIAPwas performed on DNA samples from the three patients. Molecular investigations were assessed by gene reporter expression assays in HEK cells and CRISPR-Cas9 genome editing in primary T cells.ResultsNGS ofXIAPidentified three distinct non-coding deletions in the patients that were predicted to be driven by repetitive DNA sequences. These deletions share a common region of 839 bp that encompassed the first non-coding exon ofXIAPand contained regulatory elements and marks specific of an active promoter. Moreover, we showed that among the 839 bp, the exon was transcriptionally active. Finally, deletion of the exon by CRISPR-Cas9 in primary cells reduced XIAP protein expression.ConclusionsThese results identify a key promoter sequence contained in the first non-coding exon ofXIAP. Importantly, this study highlights that sequencing of the non-coding exons that are not currently captured by WES should be considered in the genetic diagnosis when no variation is found in coding exons.
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