Department of Pharmacology and Pharmaceutical Sciences, School of Pharmacy, University of Southern California, Los Angeles, CA, United States.
Department of Ophthalmology, Roski Eye Institute, Keck School of Medicine, University of Southern California, Los Angeles, CA, United States.
Front Immunol. 2022 Mar 4;13:833254. doi: 10.3389/fimmu.2022.833254. eCollection 2022.
The tear miRNAome of the male NOD mouse, a model of ocular symptoms of Sjögren's syndrome (SS), was analyzed to identify unique miRNAs.
Male NOD mice, aged 12-14 weeks, were used to identify tear miRNAs associated with development of autoimmune dacryoadenitis. Age- and sex-matched male BALB/c mice served as healthy controls while age-matched female NOD mice that do not develop the autoimmune dacryoadenitis characteristic of SS were used as additional controls. Total RNA was isolated from stimulated tears pooled from 5 mice per sample and tear miRNAs were sequenced and analyzed. Putative miRNA hits were validated in additional mouse cohorts as well as in tears of SS patients versus patients with another form of dry eye disease, meibomian gland disease (MGD) using qRT-PCR. The pathways influenced by the validated hits were identified using Ingenuity Pathway Analysis.
In comparison to tears from both healthy (male BALB/c) and additional control (female NOD) mice, initial analy1sis identified 7 upregulated and 7 downregulated miRNAs in male NOD mouse tears. Of these, 8 were validated by RT-qPCR in tears from additional mouse cohorts. miRNAs previously implicated in SS pathology included mmu-miR-146a/b-5p, which were significantly downregulated, as well as mmu-miR-150-5p and mmu-miR-181a-5p, which were upregulated in male NOD mouse tears. All other validated hits including the upregulated miR-181b-5p and mmu-miR-203-3p, as well as the downregulated mmu-miR-322-5p and mmu-miR-503-5p, represent novel putative indicators of autoimmune dacryoadenitis in SS. When compared to tears from patients with MGD, miRNAs hsa-miR-203a-3p, hsa-miR-181a-5p and hsa-miR-181b-5p were also significantly increased in tears of SS patients.
A panel of differentially expressed miRNAs were identified in tears of male NOD mice, with some preliminary validation in SS patients, including some never previously linked to SS. These may have potential utility as indicators of ocular symptoms of SS; evaluation of the pathways influenced by these dysregulated miRNAs may also provide further insights into SS pathogenesis.
分析雄性 NOD 小鼠的泪液 miRNA 组，以鉴定独特的 miRNA。
使用 12-14 周龄雄性 NOD 小鼠鉴定与自身免疫性泪腺炎发展相关的泪液 miRNAs。年龄和性别匹配的雄性 BALB/c 小鼠作为健康对照，而不发展 SS 特征性自身免疫性泪腺炎的年龄匹配的雌性 NOD 小鼠作为额外对照。从每个样本中 5 只小鼠的刺激泪液中分离总 RNA，并对泪液 miRNA 进行测序和分析。使用 qRT-PCR 在另外的小鼠队列中以及 SS 患者与另一种干眼症，睑板腺疾病 (MGD) 患者的泪液中验证假定的 miRNA 命中。使用 Ingenuity Pathway Analysis 鉴定受验证命中影响的途径。
与来自健康对照 (雄性 BALB/c) 和另外对照 (雌性 NOD) 小鼠的泪液相比，初步分析鉴定出雄性 NOD 小鼠泪液中有 7 个上调和 7 个下调的 miRNA。其中，通过 RT-qPCR 在另外的小鼠队列中验证了 8 个 miRNA。先前与 SS 病理学相关的 miRNA 包括 mmu-miR-146a/b-5p，其表达显著下调，以及 mmu-miR-150-5p 和 mmu-miR-181a-5p，其在雄性 NOD 小鼠泪液中上调。所有其他验证的命中，包括上调的 miR-181b-5p 和 mmu-miR-203-3p，以及下调的 mmu-miR-322-5p 和 mmu-miR-503-5p，都是 SS 自身免疫性泪腺炎的新的潜在假定指标。与 MGD 患者的泪液相比，hsa-miR-203a-3p、hsa-miR-181a-5p 和 hsa-miR-181b-5p 也在 SS 患者的泪液中显著增加。
在雄性 NOD 小鼠的泪液中鉴定出一组差异表达的 miRNAs，在 SS 患者中有一些初步验证，包括一些以前从未与 SS 相关的 miRNA。它们可能具有作为 SS 眼部症状指标的潜在效用; 对这些失调的 miRNA 影响的途径进行评估也可能为 SS 的发病机制提供进一步的见解。
