Glucagon-Like Peptide-2 Stimulates S-Phase Entry of Intestinal Lgr5+ Stem Cells

Maegan E Chen; Setareh Malekian Naeini; Arjuna Srikrishnaraj; Daniel J Drucker; Zivit Fesler; Patricia L Brubaker

doi:10.1016/j.jcmgh.2022.02.011

Glucagon-Like Peptide-2 Stimulates S-Phase Entry of Intestinal Lgr5+ Stem Cells

Cell Mol Gastroenterol Hepatol. 2022;13(6):1829-1842. doi: 10.1016/j.jcmgh.2022.02.011. Epub 2022 Feb 23.

Authors

Maegan E Chen¹, Setareh Malekian Naeini¹, Arjuna Srikrishnaraj¹, Daniel J Drucker², Zivit Fesler¹, Patricia L Brubaker³

Affiliations

¹ Department of Physiology, Toronto, Ontario, Canada.
² Department of Medicine, University of Toronto, Toronto, Ontario, Canada.
³ Department of Physiology, Toronto, Ontario, Canada; Department of Medicine, University of Toronto, Toronto, Ontario, Canada. Electronic address: p.brubaker@utoronto.ca.

Abstract

Background & aims: Leucine-rich repeat-containing G-protein-coupled receptor-5 (Lgr5)+/olfactomedin-4 (Olfm4)+ intestinal stem cells (ISCs) in the crypt base are crucial for homeostatic maintenance of the epithelium. The gut hormone, glucagon-like peptide-2^1-33 (GLP-2), stimulates intestinal proliferation and growth; however, the actions of GLP-2 on the Lgr5+ ISCs remain unclear. The aim of this study was to determine whether and how GLP-2 regulates Lgr5+ ISC cell-cycle dynamics and numbers.

Methods: Lgr5-Enhanced green-fluorescent protein - internal ribosome entry site - Cre recombinase - estrogen receptor T2 (eGFP-IRES-creERT2) mice were acutely administered human Glycine² (Gly2)-GLP-2, or the GLP-2-receptor antagonist, GLP-2^3-33. Intestinal epithelial insulin-like growth factor-1-receptor knockout and control mice were treated chronically with human Gly2 (hGly2)-GLP-2. Cell-cycle parameters were determined by 5-Ethynyl-2'-deoxyuridine (EdU), bromodeoxyuridine, antibody #Ki67, and phospho-histone 3 labeling and cell-cycle gene expression.

Results: Acute hGly2-GLP-2 treatment increased the proportion of eGFP+EdU+/OLFM4+EdU+ cells by 11% to 22% (P < .05), without affecting other cell-cycle markers. hGly2-GLP-2 treatment also increased the ratio of eGFP+ cells in early to late S-phase by 97% (P < .001), and increased the proportion of eGFP+ cells entering S-phase by 218% (P < .001). hGly2-GLP-2 treatment induced jejunal expression of genes involved in cell-cycle regulation (P < .05), and increased expression of Mcm3 in the Lgr5-expressing cells by 122% (P < .05). Conversely, GLP-2^3-33 reduced the proportion of eGFP+EdU+ cells by 27% (P < .05), as well as the expression of jejunal cell-cycle genes (P < .05). Finally, chronic hGly2-GLP-2 treatment increased the number of OLFM4+ cells/crypt (P < .05), in an intestinal epithelial insulin-like growth factor-1-receptor-dependent manner.

Conclusions: These findings expand the actions of GLP-2 to encompass acute stimulation of Lgr5+ ISC S-phase entry through the GLP-2R, and chronic induction of Lgr5+ ISC expansion through downstream intestinal insulin-like growth factor-1 signaling.

Keywords: Cell Cycle; GLP-2; Intestine; Lgr5; Olfm4; Proliferation; S-Phase.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Glucagon-Like Peptide 2* / metabolism
Glucagon-Like Peptide 2* / pharmacology
Insulin-Like Growth Factor I* / metabolism
Insulin-Like Growth Factor I* / pharmacology
Intestinal Mucosa
Intestines
Mice
Receptors, G-Protein-Coupled / metabolism
Stem Cells / metabolism

Substances

Glucagon-Like Peptide 2
Lgr5 protein, mouse
Receptors, G-Protein-Coupled
Insulin-Like Growth Factor I

Abstract

Publication types

MeSH terms

Substances

Grants and funding