Transcriptomic insights into the role of the spleen in a mouse model of Wiskott-Aldrich Syndrome

Wiskott-Aldrich syndrome (WAS) is a rare X-linked primary immunodeficiency characterized by microthrombocytopenia, eczema, recurrent infections, and increased incidence of autoimmune disorders and malignancies. WAS is caused by mutations in the was gene, which is expressed exclusively in hematopoietic cells, and the spleen plays an important role in hematopoiesis and red blood cell clearance. However, to date, detailed comparative analyses of the spleen between WASp-deficient (WAS-KO) mice and WT mice, particularly at the transcriptomic level, have not been reported. Here, we investigated the differences in the transcriptomes of spleen tissues of 10-week-old WAS-KO mice. Comparison of the gene expression profiles of WAS-KO and WT mice revealed 1964 differentially expressed genes (DEGs). Among these genes, 996 DEGs were upregulated, and 968 were downregulated in WAS-KO mice. To determine the functions of the DEGs, GO and KEGG enrichment analyses were performed for significantly upregulated and downregulated DEGs, respectively. The results showed that the levels of cell senescence and apoptosis-related genes were increased, that the antigen processing and presentation mechanisms involved in the immune response were damaged, and that signal transduction processes were impaired in the spleens of WAS-KO mice. Thus, was gene deletion may lead to anemia and hemolysis-related disease, mainly due to increased osmotic fragility of red blood cells, low hemoglobin, increased bilirubin levels and increased serum ferritin. These results indicate that senescence and apoptosis of blood cells also play an important role in the occurrence of WAS. However, most studies have focused only on the immune response. Therefore, the interesting findings of this paper can provide a stronger theoretical basis for further study and help improve the treatment of WAS. We performed gene expression profiling analysis using data obtained from RNA-seq of 2 different group of WT and WAS-KO mice. Comparative gene expression profiling analysis of RNA-seq data for the spleen of WT mice and WAS-KO mice (Was -/- (129S6/SvEvTac-Wastm1Sbs/J) ).威斯科特-奥尔德里奇综合征（Wiskott-Aldrich syndrome, WAS）是一种罕见的X连锁原发性免疫缺陷病，以微小血小板性血小板减少症、湿疹、反复感染以及自身免疫紊乱和恶性肿瘤发病率升高为特征。WAS由WAS基因（was gene）突变所致，该基因仅在造血细胞中表达，而脾脏在造血功能与红细胞清除过程中发挥重要作用。然而迄今为止，尚未见针对WAS蛋白缺陷型（WASp-deficient, WAS-KO）小鼠与野生型（Wild Type, WT）小鼠脾脏的详细比较分析，尤其是转录组层面的相关研究尚未见报道。 本研究针对10周龄WAS-KO小鼠的脾脏组织转录组差异展开探究。对比WAS-KO与WT小鼠的基因表达谱，共筛选得到1964个差异表达基因（differentially expressed genes, DEGs），其中WAS-KO小鼠体内有996个基因上调、968个基因下调。为明确上述差异表达基因的功能，我们分别对显著上调和下调的DEGs进行了基因本体（Gene Ontology, GO）与京都基因与基因组百科全书（Kyoto Encyclopedia of Genes and Genomes, KEGG）富集分析。结果显示，WAS-KO小鼠脾脏内细胞衰老与凋亡相关基因表达水平升高，免疫应答过程中涉及的抗原加工提呈机制受损，且信号转导通路功能异常。此外，WAS基因缺失可能引发贫血与溶血相关疾病，这主要与红细胞渗透脆性升高、血红蛋白水平降低、胆红素水平升高以及血清铁蛋白含量增加有关。上述结果表明，血细胞衰老与凋亡在WAS的发生发展中同样发挥重要作用，但既往多数研究仅聚焦于免疫应答环节。因此，本研究的有趣发现可为后续相关研究提供更坚实的理论基础，有助于改善WAS的临床治疗。 本研究通过对两组不同的WT与WAS-KO小鼠的RNA测序（RNA sequencing, RNA-seq）数据进行分析，完成了基因表达谱检测。我们针对WT小鼠与WAS-KO小鼠（基因型为Was -/- (129S6/SvEvTac-Wastm1Sbs/J)）的脾脏组织RNA-seq数据开展了对比基因表达谱分析。