Mechanisms underlying aspirin chemoprevention of colorectal cancer remain unclear. Prior studies have been limited because of the inability of preclinical models to recapitulate human normal colon epithelium or cellular heterogeneity present in mucosal biopsies. To overcome some of these obstacles, we performed in vitro aspirin treatment of colon organoids derived from normal mucosal biopsies to reveal transcriptional networks relevant to aspirin chemoprevention. Colon organoids derived from 38 healthy individuals undergoing endoscopy were treated with 50 μmol/L aspirin or vehicle control for 72 hours and subjected to bulk RNA sequencing. Paired regression analysis using DESeq2 identified differentially expressed genes (DEG) associated with aspirin treatment. Cellular composition was determined using CIBERSORTx. Aspirin treatment was associated with 1,154 significant ( q < 0.10) DEGs prior to deconvolution. We provide replication of these findings in an independent population-based RNA-sequencing dataset of mucosal biopsies (BarcUVa-Seq), where a significant enrichment for overlap of DEGs was observed ( P < 2.2E−16). Single-cell deconvolution revealed changes in cell composition, including a decrease in transit-amplifying cells following aspirin treatment ( P = 0.01). Following deconvolution, DEGs included novel putative targets for aspirin such as TRABD2A ( q = 0.055), a negative regulator of Wnt signaling. Weighted gene co-expression network analysis identified 12 significant modules, including two that contained hubs for EGFR and PTGES2 , the latter being previously implicated in aspirin chemoprevention. In summary, aspirin treatment of patient-derived colon organoids using physiologically relevant doses resulted in transcriptome-wide changes that reveal altered cell composition and improved understanding of transcriptional pathways, providing novel insight into its chemopreventive properties. Prevention Relevance: Numerous studies have highlighted a role for aspirin in colorectal cancer chemoprevention, though the mechanisms driving this association remain unclear. We addressed this by showing that aspirin treatment of normal colon organoids diminished the transit-amplifying cell population, inhibited prostaglandin synthesis, and dysregulated expression of novel genes implicated in colon tumorigenesis.

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阿司匹林对患者来源的正常结肠类器官的全转录组体外影响


阿司匹林化学预防结直肠癌的机制仍不清楚。由于临床前模型无法重现人类正常结肠上皮或粘膜活检中存在的细胞异质性，先前的研究受到限制。为了克服其中一些障碍，我们对来自正常粘膜活检的结肠类器官进行了体外阿司匹林治疗，以揭示与阿司匹林化学预防相关的转录网络。来自接受内窥镜检查的 38 名健康个体的结肠类器官用 50 μmol/L 阿司匹林或载体对照处理 72 小时，并进行批量 RNA 测序。使用 DESeq2 的配对回归分析确定了与阿司匹林治疗相关的差异表达基因 (DEG)。使用 CIBERSORTx 确定细胞组成。在去卷积之前，阿司匹林治疗与 1,154 个显着 (q < 0.10) DEG 相关。我们在一个独立的基于群体的粘膜活检 RNA 测序数据集 (BarcUVa-Seq) 中复制了这些发现，其中观察到 DEG 重叠的显着富集 (P < 2.2E−16)。单细胞反卷积揭示了细胞组成的变化，包括阿司匹林治疗后转运放大细胞的减少（P = 0.01）。去卷积后，DEG 包括阿司匹林的新假定靶标，例如 TRABD2A (q = 0.055)，它是 Wnt 信号传导的负调节因子。加权基因共表达网络分析确定了 12 个重要模块，其中两个包含 EGFR 和 PTGES2 的中心，后者先前与阿司匹林化学预防有关。 总之，使用生理相关剂量的阿司匹林治疗患者来源的结肠类器官导致了转录组范围的变化，揭示了细胞组成的改变并提高了对转录途径的理解，为其化学预防特性提供了新的见解。预防相关性：大量研究强调了阿司匹林在结直肠癌化学预防中的作用，尽管驱动这种关联的机制仍不清楚。我们通过证明阿司匹林治疗正常结肠类器官减少了转运放大细胞群、抑制前列腺素合成以及与结肠肿瘤发生有关的新基因的表达失调来解决这个问题。