ABSTRACT
LGR5 marks resident adult epithelial stem cells at the gland base in the mouse pyloric stomach1, but the identity of the equivalent human stem cell population remains unknown owing to a lack of surface markers that facilitate its prospective isolation and validation. In mouse models of intestinal cancer, LGR5+ intestinal stem cells are major sources of cancer following hyperactivation of the WNT pathway2. However, the contribution of pyloric LGR5+ stem cells to gastric cancer following dysregulation of the WNT pathway-a frequent event in gastric cancer in humans3-is unknown. Here we use comparative profiling of LGR5+ stem cell populations along the mouse gastrointestinal tract to identify, and then functionally validate, the membrane protein AQP5 as a marker that enriches for mouse and human adult pyloric stem cells. We show that stem cells within the AQP5+ compartment are a source of WNT-driven, invasive gastric cancer in vivo, using newly generated Aqp5-creERT2 mouse models. Additionally, tumour-resident AQP5+ cells can selectively initiate organoid growth in vitro, which indicates that this population contains potential cancer stem cells. In humans, AQP5 is frequently expressed in primary intestinal and diffuse subtypes of gastric cancer (and in metastases of these subtypes), and often displays altered cellular localization compared with healthy tissue. These newly identified markers and mouse models will be an invaluable resource for deciphering the early formation of gastric cancer, and for isolating and characterizing human-stomach stem cells as a prerequisite for harnessing the regenerative-medicine potential of these cells in the clinic.
Subject(s)
Aquaporin 5/metabolism , Carcinogenesis/pathology , Neoplastic Stem Cells/pathology , Stomach Neoplasms/pathology , Stomach/pathology , Animals , Biomarkers/metabolism , Humans , Mice , Neoplastic Stem Cells/metabolism , Pylorus/pathology , Receptors, G-Protein-Coupled/metabolism , Wnt Signaling PathwayABSTRACT
The daily renewal of the corpus epithelium is fuelled by adult stem cells residing within tubular glands, but the identity of these stem cells remains controversial. Lgr5 marks homeostatic stem cells and 'reserve' stem cells in multiple tissues. Here, we report Lgr5 expression in a subpopulation of chief cells in mouse and human corpus glands. Using a non-variegated Lgr5-2A-CreERT2 mouse model, we show by lineage tracing that Lgr5-expressing chief cells do not behave as corpus stem cells during homeostasis, but are recruited to function as stem cells to effect epithelial renewal following injury by activating Wnt signalling. Ablation of Lgr5+ cells severely impairs epithelial homeostasis in the corpus, indicating an essential role for these Lgr5+ cells in maintaining the homeostatic stem cell pool. We additionally define Lgr5+ chief cells as a major cell-of-origin of gastric cancer. These findings reveal clinically relevant insights into homeostasis, repair and cancer in the corpus.
Subject(s)
Cell Proliferation , Cell Transformation, Neoplastic/metabolism , Chief Cells, Gastric/metabolism , Neoplastic Stem Cells/metabolism , Parietal Cells, Gastric/metabolism , Receptors, G-Protein-Coupled/genetics , Receptors, G-Protein-Coupled/metabolism , Regeneration , Stomach Neoplasms/metabolism , Animals , Biomarkers/metabolism , Cell Lineage , Cell Proliferation/drug effects , Cell Transformation, Neoplastic/genetics , Cell Transformation, Neoplastic/pathology , Chief Cells, Gastric/drug effects , Chief Cells, Gastric/pathology , Gene Expression Regulation , Genotype , Humans , Mice, Transgenic , Neoplastic Stem Cells/pathology , Organoids , Parietal Cells, Gastric/drug effects , Parietal Cells, Gastric/pathology , Phenotype , Regeneration/drug effects , Stomach Neoplasms/genetics , Stomach Neoplasms/pathology , Tamoxifen/toxicity , Tissue Culture Techniques , Wnt Signaling PathwayABSTRACT
The ovary surface epithelium (OSE) undergoes ovulatory tear and remodelling throughout life. Resident stem cells drive such tissue homeostasis in many adult epithelia, but their existence in the ovary has not been definitively proven. Lgr5 marks stem cells in multiple epithelia. Here we use reporter mice and single-molecule fluorescent in situ hybridization to document candidate Lgr5(+) stem cells in the mouse ovary and associated structures. Lgr5 is broadly expressed during ovary organogenesis, but becomes limited to the OSE in neonate life. In adults, Lgr5 expression is predominantly restricted to proliferative regions of the OSE and mesovarian-fimbria junctional epithelia. Using in vivo lineage tracing, we identify embryonic and neonate Lgr5(+) populations as stem/progenitor cells contributing to the development of the OSE cell lineage, as well as epithelia of the mesovarian ligament and oviduct/fimbria. Adult Lgr5(+) populations maintain OSE homeostasis and ovulatory regenerative repair in vivo. Thus, Lgr5 marks stem/progenitor cells of the ovary and tubal epithelia.
Subject(s)
Fallopian Tubes/cytology , Fallopian Tubes/metabolism , Ovary/cytology , Ovary/metabolism , Receptors, G-Protein-Coupled/genetics , Adult Stem Cells/metabolism , Animals , Animals, Newborn , Cell Lineage/genetics , Embryonic Stem Cells/metabolism , Epithelial Cells/cytology , Epithelial Cells/metabolism , Fallopian Tubes/growth & development , Female , Gene Expression , Genetic Markers , Homeostasis/genetics , Humans , In Situ Hybridization, Fluorescence , Mice , Mice, Transgenic , Models, Biological , Ovary/growth & development , Regeneration/genetics , TranscriptomeABSTRACT
Hyperhomocysteinemia (HHcy) is a metabolic disorder marked by an excess amount of the amino acid homocysteine (Hcy) in the blood stream. Hcy is a H(2)S precursor-formed from the metabolism of methionine. Elevated Hcy levels have been associated with higher blood pressure. However, the precise contribution of H(2)S to blood pressure in HHcy is not known. In the current study, we have examined a novel link between H(2)S, blood pressure and HHcy. Male Sprague-Dawley rats were injected with PAG, NaHS, L-NAME+PAG and saline. HHcy condition was induced by providing methionine (1 g/kg) in drinking water for 8 weeks. After 8 weeks, plasma Hcy and H(2)S were measured. The treated rats were anaesthetized with a mixture of ketamine hydrochloride and medetomidine. Blood pressures were measured by intra-carotid artery catheterization and to further investigate the immediate effect of NO and H(2)S, exogenous drugs namely NaHS, SNP, Ach and NA were administered. Plasma Hcy levels were higher in HHcy groups and this group exhibited hypertension. We observed high blood pressure at low levels of H(2)S and vice versa. Endogenous H(2)S in HHcy condition facilitated a mild decrease in MAP (Mean Arterial Pressure). Exogenous SNP (NO donor) showed a greater pressure decrease in HHcy group. The underlying mechanism is yet to be exploited. High levels of Hcy play an important role in the pathogenesis of hypertension. The results suggest that both endogenous and exogenous H(2)S may play a vital role in regulating blood pressure in HHcy.