Transcriptomic Response to Calcium in Normal Colon Organoids is Impacted by Colon Location and Sex.
Transcriptomic Response to Calcium in Normal Colon Organoids is Impacted by Colon Location and Sex.
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DOI：
10.1158/1940-6207.capr-22-0068
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发表时间：
2022-10-04
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观察性研究表明，补充钙可以预防结直肠癌。分层分析表明，这种保护作用可能因解剖亚位点和性别而异，但这些假设很难通过实验进行检验。在这里，我们将来自无关受试者（18名女性，18名男性）的正常结肠活检（21个右结肠，15个左结肠）的36个患者来源的类器官线暴露于中等（1.66 mM）或高（5.0 mM）浓度的钙72小时。我们进行了批量 RNA 测序来测量基因表达，并使用 CIBERSORTx 中的单细胞反卷积推断细胞组成。我们使用 DESeq2 中的广义线性模型测试了基因表达的显着差异。暴露于较高水平的钙与细胞组成的变化相关（P<0.05），最显着的是杯状细胞增加和干细胞数量减少，以及 485 个基因的差异表达（FDR<0.05）。我们发现，其中 40 个差异表达基因映射到通过结直肠癌全基因组关联研究确定的基因组位点，这表明补钙和遗传性结直肠癌风险之间存在潜在的生物学重叠。分层分析发现，来自右侧结肠和男性受试者的结肠类器官中的差异表达基因多于来自左侧结肠和女性受试者的结肠类器官。我们在一组匹配的左右结肠类器官 (n=4) 中使用定量 PCR 证实了其中一个基因 HSD17B2 存在更强的右侧效应。通过将我们的发现与遗传数据联系起来，我们提供了关于营养和遗传因素如何相互作用影响结直肠癌风险的新见解。
Observational studies indicate that calcium supplementation may protect against colorectal cancer. Stratified analyses suggest that this protective effect may differ based on anatomic subsite and sex, but these hypotheses have been difficult to test experimentally. Here, we exposed 36 patient-derived organoid lines derived from normal colon biopsies (21 right colon, 15 left colon) of unrelated subjects (18 female, 18 male) to moderate (1.66 mM) or high (5.0 mM) concentrations of calcium for 72 hours. We performed bulk RNA-sequencing to measure gene expression, and cell composition was inferred using single-cell deconvolution in CIBERSORTx. We tested for significant differences in gene expression using generalized linear models in DESeq2. Exposure to higher levels of calcium was associated with changes in cell composition (P<0.05), most notably increased goblet and reduced stem cell populations, and differential expression of 485 genes (FDR<0.05). We found that 40 of these differentially expressed genes mapped to genomic loci identified through colorectal cancer genome-wide association studies, suggesting a potential biologic overlap between calcium supplementation and inherited colorectal cancer risk. Stratified analyses identified more differentially expressed genes in colon organoids derived from right sided colon and male subjects than those derived from left sided colon and female subjects. We confirmed the presence of a stronger right-sided effect for one of these genes, HSD17B2 using quantitative PCR in a subset of matched right and left colon organoids (n=4). By relating our findings to genetic data, we provide new insights into how nutritional and genetic factors may interact to influence CRC risk.