Antonczyk Aleksandra, Kluzek Katarzyna, Herbich Natalia, Boroujeni Mahdi Eskandarian, Krist Bart, Wronka Dorota, Karlik Anna, Przybyl Lukasz, Plewinski Adam, Wesoly Joanna, Bluyssen Hans A R
Human Molecular Genetics Research Unit, Institute of Molecular Biology and Biotechnology, Faculty of Biology, Adam Mickiewicz University, Poznan, Poland.
Laboratory of Mammalian Model Organisms, Institute of Bioorganic Chemistry, Polish Academy of Sciences, Poznan, Poland.
Front Pharmacol. 2025 Jan 7;15:1471182. doi: 10.3389/fphar.2024.1471182. eCollection 2024.
Cardiovascular diseases (CVDs) include atherosclerosis, which is an inflammatory disease of large and medium vessels that leads to atherosclerotic plaque formation. The key factors contributing to the onset and progression of atherosclerosis include the pro-inflammatory cytokines interferon (IFN)α and IFNγ and the pattern recognition receptor (PRR) Toll-like receptor 4 (TLR4). Together, they trigger the activation of IFN regulatory factors (IRFs) and signal transducer and activator of transcription (STAT)s. Based on their promoting role in atherosclerosis, we hypothesized that the inhibition of pro-inflammatory target gene expression through multi-IRF inhibitors may be a promising strategy to treat CVDs. Using comparative docking of multiple IRF-DNA-binding domain (DBD) models on a multi-million natural compound library, we identified the novel multi-IRF inhibitor, ALEKSIN. This compound targets the DBD of IRF1, IRF2, and IRF8 with the same affinity and simultaneously inhibits the expression of multiple IRF target genes in human microvascular endothelial cells (HMECs) in response to IIFNα and IFNγ. Under the same conditions, ALEKSIN also inhibited the phosphorylation of STATs, potentially through low-affinity STAT-SH2 binding but with lower potency than the known multi-STAT inhibitor STATTIC. This was in line with the common inhibition of ALEKSIN and STATTIC observed on the genome-wide expression of pro-inflammatory IRF/STAT/NF-κB target genes, as well as on the migration of HMECs. Finally, we identified a novel signature of 46 ALEKSIN and STATTIC commonly inhibited pro-atherogenic target genes, which was upregulated in atherosclerotic plaques in the aortas of high-fat diet-fed ApoEKO mice and associated with inflammation, proliferation, adhesion, chemotaxis, and response to lipids. Interestingly, the majority of these genes could be linked to macrophage subtypes present in aortic plaques in HFD-fed LDLR-KO mice. Together, this suggests that ALEKSIN represents a novel class of multi-IRF inhibitors, which inhibits IRF-, STAT-, and NF-κB-mediated transcription and could offer great promise for the treatment of CVDs. Furthermore, the ALEKSIN and STATTIC commonly inhibited pro-inflammatory gene signature could help monitor plaque progression during experimental atherosclerosis.
心血管疾病（CVDs）包括动脉粥样硬化，它是一种大中血管的炎症性疾病，会导致动脉粥样硬化斑块形成。促成动脉粥样硬化发生和发展的关键因素包括促炎细胞因子干扰素（IFN）α和IFNγ以及模式识别受体（PRR）Toll样受体4（TLR4）。它们共同触发干扰素调节因子（IRFs）以及信号转导和转录激活因子（STATs）的激活。基于它们在动脉粥样硬化中的促进作用，我们推测通过多IRF抑制剂抑制促炎靶基因表达可能是治疗心血管疾病的一种有前景的策略。通过在数百万种天然化合物库上对多个IRF-DNA结合域（DBD）模型进行比较对接，我们鉴定出了新型多IRF抑制剂ALEKSIN。该化合物以相同亲和力靶向IRF1、IRF2和IRF8的DBD，并同时抑制人微血管内皮细胞（HMECs）中多个IRF靶基因对IIFNα和IFNγ的响应表达。在相同条件下，ALEKSIN还抑制了STATs的磷酸化，可能是通过低亲和力的STAT-SH2结合，但效力低于已知的多STAT抑制剂STATTIC。这与ALEKSIN和STATTIC在促炎IRF/STAT/NF-κB靶基因的全基因组表达以及HMECs迁移上的共同抑制作用一致。最后，我们鉴定出了46个ALEKSIN和STATTIC共同抑制的促动脉粥样硬化靶基因的新特征，这些基因在高脂饮食喂养的ApoEKO小鼠主动脉粥样硬化斑块中上调，并与炎症、增殖、黏附、趋化作用以及对脂质的反应相关。有趣的是，这些基因中的大多数可能与高脂饮食喂养的LDLR-KO小鼠主动脉斑块中存在的巨噬细胞亚型有关。总之，这表明ALEKSIN代表了一类新型的多IRF抑制剂，它抑制IRF、STAT和NF-κB介导的转录，可能为心血管疾病的治疗带来巨大希望。此外，ALEKSIN和STATTIC共同抑制的促炎基因特征有助于在实验性动脉粥样硬化过程中监测斑块进展。
