Najjar Rayan, Wang Xiaoxing, Pineda Jose Mario Bello, Alessi Hugh, Bays Alison, Bradley Robert K, Jarvis James N, Mustelin Tomas
University of Washington, Seattle.
University of Washington, Seattle, and Fred Hutchinson Cancer Research Center, Seattle, Washington.
ACR Open Rheumatol. 2025 Jan;7(1):e11770. doi: 10.1002/acr2.11770.
To test whether messenger RNA (mRNA) splicing is altered in neutrophils from patients with systemic lupus erythematosus (SLE) and can produce neoantigens.
RNA sequencing of neutrophils from patients with SLE (n = 15) and healthy donors (n = 12) were analyzed for mRNA splicing using the RiboSplitter pipeline, an event-focused tool based on SplAdder with subsequent translation and protein domain annotation. RNA sequencing from SARS-CoV2-infected individuals was used as an additional comparator.
Neutrophils from patients with SLE contained 521 statistically significant altered mRNA splicing events compared with healthy donor neutrophils, many of them affecting important immunologic pathways, myeloid function, transcription factors, and proteins involved in mRNA splicing. A subset of splicing events were only present in SLE samples, and some of them occurred at unannotated splice acceptor or donor sites. Two patients were particularly rich in such events. Only a small number of dysregulated splicing events were more pronounced in patients with active disease or with high type I interferons but were not detected in SARS-CoV2-infected individuals, who also had high type I interferons. Besides causing a range of structural changes, 80 mRNA splice variants exclusive to SLE were predicted to translate into novel amino acid sequences. Peptides derived from these novel amino acid sequences were predicted to bind to the individual patients' class I and II major histocompatibility complex molecules with high affinity.
We conclude that aberrant mRNA splicing in SLE has the potential to affect both the function of granulocytes and to generate novel autoantigens.
检测系统性红斑狼疮（SLE）患者中性粒细胞中的信使核糖核酸（mRNA）剪接是否发生改变以及是否能产生新抗原。
使用RiboSplitter流程对SLE患者（n = 15）和健康供者（n = 12）的中性粒细胞进行RNA测序，以分析mRNA剪接情况。RiboSplitter流程是一种基于SplAdder的聚焦事件工具，随后进行翻译和蛋白质结构域注释。将来自感染严重急性呼吸综合征冠状病毒2（SARS-CoV2）个体的RNA测序用作额外的对照。
与健康供者的中性粒细胞相比，SLE患者的中性粒细胞含有521个具有统计学意义的mRNA剪接改变事件，其中许多影响重要的免疫途径、髓系功能、转录因子以及参与mRNA剪接的蛋白质。一部分剪接事件仅存在于SLE样本中，其中一些发生在未注释的剪接受体或供体位点。两名患者的此类事件尤为丰富。只有少数失调的剪接事件在疾病活动期患者或I型干扰素水平高的患者中更为明显，但在同样具有高I型干扰素水平的SARS-CoV2感染个体中未检测到。除了引起一系列结构变化外，预计SLE特有的80种mRNA剪接变体可翻译成新的氨基酸序列。源自这些新氨基酸序列的肽预计会以高亲和力与个体患者的I类和II类主要组织相容性复合体分子结合。
我们得出结论，SLE中异常的mRNA剪接有可能影响粒细胞功能并产生新的自身抗原。
