Department of Public Health Sciences, Center for Public Health Genomics, University of Virginia, Charlottesville, VA, USA.
Department of Surgery, Center for Public Health Genomics, University of Virginia, Charlottesville, VA, USA.
Sci Rep. 2021 Jan 11;11(1):432. doi: 10.1038/s41598-020-80240-1.
Alcohol is a consistently identified risk factor for colon cancer. However, the molecular mechanism underlying its effect on normal colon crypt cells remains poorly understood. We employed RNA-sequencing to asses transcriptomic response to ethanol exposure (0.2% vol:vol) in 3D organoid lines derived from healthy colon (n = 34). Paired regression analysis identified 2,162 differentially expressed genes in response to ethanol. When stratified by colon location, a far greater number of differentially expressed genes were identified in organoids derived from the left versus right colon, many of which corresponded to cell-type specific markers. To test the hypothesis that the effects of ethanol treatment on colon organoid populations were in part due to differential cell composition, we incorporated external single cell RNA-sequencing data from normal colon biopsies to estimate cellular proportions following single cell deconvolution. We inferred cell-type-specific changes, and observed an increase in transit amplifying cells following ethanol exposure that was greater in organoids from the left than right colon, with a concomitant decrease in more differentiated cells. If this occurs in the colon following alcohol consumption, this would lead to an increased zone of cells in the lower crypt where conditions are optimal for cell division and the potential to develop mutations.
酒精是结肠癌的一个明确的危险因素。然而，其对正常结肠隐窝细胞影响的分子机制仍知之甚少。我们采用 RNA 测序技术，在源自健康结肠的 3D 类器官系（n = 34）中评估乙醇暴露（0.2%体积比）的转录组反应。配对回归分析鉴定出 2162 个对乙醇有差异表达的基因。按结肠位置分层，源自左结肠的类器官比源自右结肠的类器官鉴定出更多数量的差异表达基因，其中许多与细胞类型特异性标志物相对应。为了验证乙醇处理对结肠类器官群体的影响部分归因于细胞组成的差异的假设，我们整合了来自正常结肠活检的外部单细胞 RNA 测序数据，以估计单细胞去卷积后细胞的比例。我们推断出细胞类型特异性的变化，并观察到乙醇暴露后过渡扩增细胞增加，左结肠的增加幅度大于右结肠，而分化程度更高的细胞则相应减少。如果这种情况发生在饮酒后的结肠中，这将导致下部隐窝中细胞分裂条件最佳且有潜力发生突变的区域增加。
