Fall of PARP3 restrains Lgr5⁺ intestinal stem cells proliferation and mucosal renovation in intestinal aging

The regeneration ability of intestinal epithelium is degenerated in aging. The determining factor is leucine-rich repeat-containing G-protein-coupled receptor 5-positive intestinal stem cells (Lgr5⁺ ISCs). Lgr5-EGFP (enhanced green fluorescence protein) knock-in in transgenic mice at three different ages (young group: 3-6 months; middle group: 12-14 months; old group: 22-24 months) were used to examined Lgr5⁺ ISCs at three different timepoints. The jejunum samples were collected for histology, immunofluorescence analysis, western blotting and PCR. In tissue, crypt depth, proliferating cells and Lgr5⁺ ISC numbers were increased in the middle group (12-14 months) and decreased in the old group (22-24 months). The number of proliferating Lgr5⁺ ISCs gradually decreased as the mice aged. In organoids, the budding number, projected area, and Lgr5⁺ ISC ratio decreased as the mice aged. The gene expression of poly (ADP-ribose) polymerase 3 (Parp3) and the protein expression of PARP3 were increased in the middle- and old-aged groups. PARP3 inhibitors slowed organoid growth in the middle group. In conclusion, PARP3 is upregulated in aging, and the inhibition of PARP3 reduces the proliferation of aging Lgr5⁺ ISCs.