Effect of depletion of C15ORF48/miR-147 on gene expression in primary colonocytes [HCT116]. Effect of depletion of C15ORF48/miR-147 on gene expression in primary colonocytes [HCT116]

Chronic inflammation is epidemiologically linked to the pathogenesis of gastrointestinal diseases, including inflammatory bowel disease (IBD) and colorectal cancer (CRC). However, our understanding of the molecular mechanisms controlling gut inflammation remains insufficient, hindering the development of targeted therapies for IBD and CRC. In this study, we uncovered C15ORF48/miR-147 as a novel negative regulator of gut inflammation, operating through the modulation of epithelial cell metabolism. C15ORF48/miR-147 encodes two molecular products, C15ORF48 protein and miR-147-3p microRNA, which are predominantly expressed in intestinal epithelium. C15ORF48/miR-147 ablation leads to gut dysbiosis and exacerbates chemically-induced colitis in mice. C15ORF48 and miR-147-3p work together to suppress colonocyte metabolism and inflammatory responses, and thus bridging mitochondrial metabolism and inflammation. Overall design: To define the molecular mechanism by which C15ORF48/miR 147 controls gut inflammation, we performed global transcriptome analysis of purified C15ORF48/miR 147-sufficient and deficient colonocytes from unchallenged and DSS challenged mice using RNA-seq.

慢性炎症与包括炎症性肠病（Inflammatory Bowel Disease, IBD）、结直肠癌（Colorectal Cancer, CRC）在内的胃肠道疾病的发病机制存在流行病学关联。然而，当前学界对调控肠道炎症的分子机制的认知仍存在不足，这阻碍了针对IBD与CRC的靶向治疗开发。
本研究中，我们鉴定出C15ORF48/miR-147作为一类新型肠道炎症负调控因子，其通过调控上皮细胞代谢发挥功能。C15ORF48/miR-147可编码两种分子产物：C15ORF48蛋白与miR-147-3p 微小RNA（microRNA），二者主要在肠上皮细胞中表达。敲除C15ORF48/miR-147会导致小鼠肠道菌群失调，并加重化学诱导型结肠炎。C15ORF48与miR-147-3p协同抑制结肠上皮细胞的代谢与炎症反应，从而搭建起线粒体代谢与炎症之间的桥梁。
整体实验设计：为阐明C15ORF48/miR-147调控肠道炎症的分子机制，我们采用RNA-seq技术，对未经造模及经葡聚糖硫酸钠（Dextran Sulfate Sodium, DSS）造模的小鼠体内纯化得到的C15ORF48/miR-147充足型与缺陷型结肠上皮细胞开展了全局转录组分析。