Sampson Leesa L, Davis Ashley K, Grogg Matthew W, Zheng Yi
Division of Experimental Hematology and Cancer Biology, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio, USA.
Division of Experimental Hematology and Cancer Biology, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio, USA
FASEB J. 2016 Mar;30(3):1263-75. doi: 10.1096/fj.15-278606. Epub 2015 Nov 30.
Intestinal stem cells (ISCs) drive small intestinal epithelial homeostasis and regeneration. Mechanistic target of rapamycin (mTOR) regulates stem and progenitor cell metabolism and is frequently dysregulated in human disease, but its physiologic functions in the mammalian small intestinal epithelium remain poorly defined. We disrupted the genes mTOR, Rptor, Rictor, or both Rptor and Rictor in mouse ISCs, progenitors, and differentiated intestinal epithelial cells (IECs) using Villin-Cre. Mutant tissues and wild-type or heterozygous littermate controls were analyzed by histologic immunostaining, immunoblots, and proliferation assays. A total of 10 Gy irradiation was used to injure the intestinal epithelium and induce subsequent crypt regeneration. We report that mTOR supports absorptive enterocytes and secretory Paneth and goblet cell function while negatively regulating chromogranin A-positive enteroendocrine cell number. Through additional Rptor, Rictor, and Rptor/Rictor mutant mouse models, we identify mechanistic target of rapamycin complex 1 as the major IEC regulatory pathway, but mechanistic target of rapamycin complex 2 also contributes to ileal villus maintenance and goblet cell size. Homeostatic adult small intestinal crypt cell proliferation, survival, and canonical wingless-int (WNT) activity are not mTOR dependent, but Olfm4(+) ISC/progenitor population maintenance and crypt regeneration postinjury require mTOR. Overall, we conclude that mTOR regulates multiple IEC lineages and promotes stem and progenitor cell activity during intestinal epithelium repair postinjury.
肠道干细胞(ISCs)驱动小肠上皮的稳态和再生。雷帕霉素机制性靶标(mTOR)调节干细胞和祖细胞的代谢,并且在人类疾病中经常失调,但其在哺乳动物小肠上皮中的生理功能仍不清楚。我们使用Villin-Cre在小鼠的肠道干细胞、祖细胞和分化的肠上皮细胞(IECs)中破坏了mTOR、Rptor、Rictor或Rptor和Rictor这两个基因。通过组织学免疫染色、免疫印迹和增殖测定分析突变组织以及野生型或杂合子同窝对照。使用总共10 Gy的辐射损伤肠上皮并诱导随后的隐窝再生。我们报告称,mTOR支持吸收性肠上皮细胞以及分泌性潘氏细胞和杯状细胞的功能,同时对嗜铬粒蛋白A阳性肠内分泌细胞的数量起负调节作用。通过构建额外的Rptor、Rictor和Rptor/Rictor突变小鼠模型,我们确定雷帕霉素机制性靶标复合物1是主要的IEC调节途径,但雷帕霉素机制性靶标复合物2也有助于回肠绒毛的维持和杯状细胞的大小。成年小肠隐窝细胞的稳态增殖、存活和经典无翅型(WNT)活性不依赖于mTOR,但Olfm4(+)肠道干细胞/祖细胞群体的维持以及损伤后的隐窝再生需要mTOR。总体而言,我们得出结论,mTOR调节多种IEC谱系,并在损伤后肠道上皮修复过程中促进干细胞和祖细胞的活性。