Functional impact of IRF4 mutations on transcription in patient-derived B-EBV cells [RNA-seq]. Functional impact of IRF4 mutations on transcription in patient-derived B-EBV cells [RNA-seq]

An IRF4 de novo mutation affecting the DNA binding domain of encoded IRF4 protein (mutDBD) was identified in a patient presenting with combined immunodeficiency. The patient exhibited profound susceptibility to opportunistic infections notably Pneumocystis jirovecii and humoral immunodeficiency caused by a failure of terminal B cell differentiation. A heterozygous IRF4 missense variant resulting in a phenylalanine-to-leucine replacement within the interferon activation domain of the encoded IRF4 protein (mutIAD) was identified in three patients from a multigenerational family suffering from a novel autosomal dominant disease predominantly presenting as a hypogammaglobulinemia with recurrent infections. In this experiment we aimed to investigate the effect of the two different mutations on IRF4 regulated transcription. Overall design: RNA-seq data in EBV-B cells of individuals heterozygous for IRF4 mutations and healthy homozygous WT individuals

本研究在1名表现为联合免疫缺陷的患者体内，发现了1个影响编码IRF4蛋白DNA结合域的IRF4新生突变（mutDBD）。该患者对机会性感染的易感性显著升高，尤其易感染耶氏肺孢子菌（Pneumocystis jirovecii），同时存在因终末B细胞分化障碍引发的体液免疫缺陷。

本研究同时在一个多代家系的3名患者体内，鉴定出1个杂合型IRF4错义变异：该变异导致编码IRF4蛋白的干扰素激活域内发生苯丙氨酸向亮氨酸的替换，暂命名为mutIAD。这3名患者均罹患一种新型常染色体显性疾病，其主要临床表现为低丙种球蛋白血症伴复发性感染。

本实验旨在探究这两种不同突变对IRF4调控的转录过程的影响。
实验整体设计：采集IRF4突变杂合携带者与健康纯合野生型（WT）个体的EB病毒转化B细胞（EBV-B cells）的RNA测序（RNA-seq）数据。