Vanderbilt University Medical Center, Nashville, Tennessee.
Vanderbilt University Medical Center and Tennessee Valley Healthcare System, U.S. Department of Veterans Affairs, Nashville.
Arthritis Rheumatol. 2024 May;76(5):684-695. doi: 10.1002/art.42782. Epub 2024 Jan 24.
High-density lipoprotein (HDL) has well-characterized anti-atherogenic cholesterol efflux and antioxidant functions. Another function of HDL uncharacterized in rheumatoid arthritis (RA) is its ability to transport microRNAs (miRNAs) between cells and thus alter cellular function. The study's purpose was to determine if HDL-miRNA cargo is altered and affects inflammation in RA.
HDL-microRNAs were characterized in 30 RA and 30 control participants by next generation sequencing and quantitative polymerase chain reaction. The most abundant differentially expressed miRNA was evaluated further. The function of miR-1246 was assessed by miRNA mimics, antagomiRs, small interfering RNA knockdown, and luciferase assays. Monocyte-derived macrophages were treated with miR-1246-loaded HDL and unmodified HDL from RA and control participants to measure delivery of miR-1246 and its effect on interleukin-6 (IL-6).
The most abundant miRNA on HDL was miR-1246; it was significantly enriched two-fold on HDL from RA versus control participants. HDL-mediated miR-1246 delivery to macrophages significantly increased IL6 expression 43-fold. miR-1246 delivery significantly decreased DUSP3 1.5-fold and DUSP3 small interfering RNA knockdown increased macrophage IL6 expression. Luciferase assay indicated DUSP3 is a direct target of miR-1246. Unmodified HDL from RA delivered 1.6-fold more miR-1246 versus control participant HDL. Unmodified HDL from both RA and control participants attenuated activated macrophage IL6 expression, but this effect was significantly blunted in RA so that IL6 expression was 3.4-fold higher after RA versus control HDL treatment.
HDL-miR-1246 was increased in RA versus control participants and delivery of miR-1246 to macrophages increased IL-6 expression by targeting DUSP3. The altered HDL-miRNA cargo in RA blunted HDL's anti-inflammatory effect.
高密度脂蛋白（HDL）具有明确的抗动脉粥样硬化胆固醇外排和抗氧化功能。HDL 在类风湿关节炎（RA）中尚未明确的另一个功能是其在细胞间运输 microRNAs（miRNAs）的能力，从而改变细胞功能。本研究旨在确定 HDL-miRNA 载运物是否发生改变并影响 RA 中的炎症。
通过下一代测序和定量聚合酶链反应对 30 名 RA 患者和 30 名对照参与者的 HDL 微小 RNA 进行了表征。进一步评估了最丰富的差异表达 miRNA。通过 miRNA 模拟物、反义寡核苷酸、小干扰 RNA 敲低和荧光素酶测定评估 miR-1246 的功能。用负载 miR-1246 的 HDL 和来自 RA 和对照参与者的未修饰 HDL 处理单核细胞衍生的巨噬细胞，以测量 miR-1246 的递呈及其对白细胞介素 6（IL-6）的影响。
HDL 上最丰富的 miRNA 是 miR-1246；它在 RA 患者的 HDL 上富集了两倍。HDL 介导的 miR-1246 向巨噬细胞的递呈使 IL6 表达增加了 43 倍。miR-1246 递呈使 DUSP3 减少 1.5 倍，DUSP3 小干扰 RNA 敲低增加了巨噬细胞 IL6 表达。荧光素酶测定表明 DUSP3 是 miR-1246 的直接靶标。与对照参与者的 HDL 相比，RA 的未修饰 HDL 递呈了 1.6 倍的 miR-1246。来自 RA 和对照参与者的未修饰 HDL 均减弱了活化巨噬细胞的 IL6 表达，但 RA 中的这种作用明显减弱，因此在用 RA 与对照 HDL 处理后，IL6 表达增加了 3.4 倍。
与对照参与者相比，RA 患者的 HDL-miR-1246 增加，向巨噬细胞递呈 miR-1246 通过靶向 DUSP3 增加了 IL-6 表达。RA 中改变的 HDL-miRNA 载运物削弱了 HDL 的抗炎作用。
