Department of Clinical Laboratory, Peking Union Medical College Hospital, Peking Union Medical College, Chinese Academy of Medical Sciences, Beijing 100730, P. R. China.
Department of Clinical Laboratory, West China Second University Hospital, Sichuan University, Chengdu 610041, P. R. China.
J Proteome Res. 2023 Oct 6;22(10):3254-3263. doi: 10.1021/acs.jproteome.3c00268. Epub 2023 Aug 28.
Systemic sclerosis (SSc) is a systemic autoimmune disorder that leads to vasculopathy and tissue fibrosis. A lack of reliable biomarkers has been a challenge for clinical diagnosis of the disease. We employed a protein array-based approach to identify and validate SSc-specific autoantibodies. Phase I involved profiled autoimmunity using human proteome microarray (HuProt arrays) with 90 serum samples: 40 patients with SSc, 30 patients diagnosed with autoimmune diseases, and 20 healthy subjects. In Phase II, we constructed a focused array with candidates identified antigens and used this to profile a much larger cohort comprised of serum samples. Finally, we used a western blot analysis to validate the serum of validated proteins with high signal values. Bioinformatics analysis allowed us to identify 113 candidate autoantigens that were significantly associated with SSc. This two-phase strategy allowed us to identify and validate anti-small nuclear ribonucleoprotein polypeptide A (SNRPA) as a novel SSc-specific serological biomarker. The observed positive rate of anti-SNRPA antibody in patients with SSc was 11.25%, which was significantly higher than that of any disease control group (3.33%) or healthy controls (1%). In conclusion, anti-SNRPA autoantibody serves as a novel biomarker for SSc diagnosis and may be promising for clinical applications.
系统性硬化症(SSc)是一种系统性自身免疫性疾病,可导致血管病变和组织纤维化。缺乏可靠的生物标志物一直是临床诊断该疾病的挑战。我们采用基于蛋白质阵列的方法来鉴定和验证 SSc 特异性自身抗体。第一阶段涉及使用人类蛋白质组微阵列(HuProt 阵列)对自身免疫进行分析,该阵列使用了 90 份血清样本:40 名 SSc 患者、30 名自身免疫性疾病患者和 20 名健康受试者。在第二阶段,我们使用鉴定出的候选抗原构建了一个聚焦阵列,并使用该阵列对包含更多血清样本的队列进行了分析。最后,我们使用 Western blot 分析对具有高信号值的验证蛋白的血清进行了验证。生物信息学分析使我们能够鉴定出 113 种与 SSc 显著相关的候选自身抗原。这种两阶段策略使我们能够鉴定和验证抗小核核糖核蛋白多肽 A(SNRPA)作为一种新型的 SSc 特异性血清学生物标志物。在 SSc 患者中观察到抗 SNRPA 抗体的阳性率为 11.25%,明显高于任何疾病对照组(3.33%)或健康对照组(1%)。总之,抗 SNRPA 自身抗体可作为 SSc 诊断的新型生物标志物,可能具有广阔的临床应用前景。