Lgr5-expressing stem cells are not the cells of origin of pyloric neuroendocrine carcinomas in mice.
In intestinal and pyloric epithelia, leucine-rich repeat-containing G protein-coupled receptor 5 (Lgr5)-expressing cells represent long-lived adult stem cells that give rise to all epithelial cell types, including endocrine cells. Ablation of the Apc gene in Lgr5-expressing cells leads to intestinal and pyloric adenomas. To assess whether all epithelial tumours of the gastrointestinal tract are derived from LGR5-positive stem cells, we crossed Lgr5-EGFP-IRES-creER(T2) mice, which express EGFP and Cre recombinase driven by the Lgr5 promoter, with CEA424-SV40-TAg mice, which develop pyloric neuroendocrine carcinomas of epithelial origin. In 19 day-old mice, single SV40 T antigen (TAg)-positive cells were identified preferentially at the the bases of pyloric glands, close to the stem cell compartment. However, contrary to previous publications describing subpopulations of LGR5-positive cells in gastrointestinal neoplasia, we could not detect Lgr5-EGFP-positive tumour cells in malignant lesions. The lack of expression of the Wnt target gene Lgr5 is probably not caused by suppression of Wnt signalling by TAg, since β-catenin-mediated Wnt signalling, as measured by the TOPflash assay, was not inhibited. To determine the cellular origin of CEA424-SV40-TAg tumours, we performed tracing experiments using Lgr5-EGFP-IRES-creERT2:CEA424-SV40-TAg:ROSA26-tdRFP mice. Following tamoxifen induction, it was possible to efficiently trace the progeny of Lgr5-expressing cells in gastrointestinal tissue via red fluorescent protein (RFP) expression. No RFP-positive tumour cells were detected, even when RFP gene activation occurred in 7 day-old mice well before the appearance of TAg-positive tumour cells. Hence, we conclude that Lgr5-expressing stem cells probably do not constitute the cells of origin in CEA424-SV40-TAg mice. Consequently, not all epithelial tumours in the pyloric region are initiated by transformation of LGR5-positive stem cells. Thus, additional long-lived LGR5-negative stem cells or progenitor cells with a low turnover rate might exist in the pyloric region, which could give rise to tumours.
Tumour Immunology Laboratory, LIFE Centre, University Hospital, Ludwig-Maximilians University, Munich, Germany.
Elena
E
Vetter
Tumour Immunology Laboratory, LIFE Centre, University Hospital, Ludwig-Maximilians University, Munich, Germany.
Mira
M
Kronast
Tumour Immunology Laboratory, LIFE Centre, University Hospital, Ludwig-Maximilians University, Munich, Germany.
Mariana
M
Tölge
Tumour Immunology Laboratory, LIFE Centre, University Hospital, Ludwig-Maximilians University, Munich, Germany.
Wolfgang
W
Zimmermann
D000818:Animals; D018278:Carcinoma, Neuroendocrine / Q000473:pathology*; D019070:Cell Lineage; D004195:Disease Models, Animal; D005455:Fluorescent Antibody Technique; D007150:Immunohistochemistry; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D014411:Neoplastic Stem Cells / Q000378:metabolism / Q000473:pathology*; D011708:Pylorus / Q000473:pathology; D043562:Receptors, G-Protein-Coupled / Q000096:biosynthesis*; D013274:Stomach Neoplasms / Q000473:pathology*
C493597:Lgr5 protein, mouse; D043562:Receptors, G-Protein-Coupled
C57BL/6 mice; LGR5; TAg; carcinoma; cell of origin; enteroendocrine; gastric; neuroendocrine; pylorus