LSD1 represses a neonatal/reparative gene program in adult intestinal epithelium

Abstract

30Intestinal epithelial homeostasis is maintained by adult intestinal stem cells, which, alongside 31 Paneth cells, appear after birth in the neonatal period. We aimed to identify new regulators of 32 neonatal intestinal epithelial development by testing a small library of epigenetic modifier 33 inhibitors in Paneth cell-skewed organoid cultures. We found that Lysine-specific 34 demethylase 1A (Kdm1a/Lsd1) is absolutely required for Paneth cell differentiation. Lsd1-35 deficient crypts, devoid of Paneth cells, are still able to form organoids without a requirement 36 of exogenous or endogenous Wnt. Mechanistically, we find that LSD1 represses genes that 37 are normally expressed in fetal and neonatal epithelium. This gene profile is similar to what is 38 seen in repairing epithelium, and indeed, we find that Lsd1-deficient epithelium has superior 39 regenerative capacities after irradiation injury. In summary, we found an important regulator 40 of neonatal intestinal development and identified a druggable target to reprogram intestinal 41 epithelium towards a reparative state. 42indicate PC + crypts based on presence of granular cells, asterisks indicate PCcrypts. D, Brightfield images and

Intestinal-epithelial LSD1 controls cytoskeletal-mediated cell identity including goblet cell effector responses required for gut inflammatory and infectious diseases

“…Yui et al identified that YAP/TAZ-mediated reprogramming of intestinal epithelium into a repair state is important in this disease model (Yui et al, 2018), and we found that Lsd1-deficient epithelium matches the phenotype of reparative epithelium ( Fig. 1, and (Zwiggelaar et al, 2020). We thus wanted to test whether Lsd1-deficient epithelium would be protective upon inflammatory damage, like it is upon damage induced by irradiation.…”

“…LSD1 can demethylate mono or dimethylated Histone 3 Lysine 4 (H3K4me1/2) that normally mark active or poised enhancers (Shi et al, 2004). Importantly, we previously found that LSD1-deficient small intestinal epithelium is in a continuous state of repair (Zwiggelaar et al, 2020).…”

“…We have recently shown that inhibition or deletion of the histone demethylase LSD1 renders the small intestinal epithelium in a continuous state of repair, and this state is beneficial for recovery after irradiation injury (Zwiggelaar et al, 2020). We generated mice that lack Lsd1 in the intestinal epithelium specifically by crossing Villin-Cre mice with Lsd1 f/f mice (conditional knock-out (cKO) mice) and found that these mice have a near complete deletion of Lsd1 in small intestinal epithelium (Zwiggelaar et al, 2020). In the current study, we focus on the colon, and here we also find crypts still expressing LSD1 in cKO mice (Fig.…”

“…Finally, IL-22 is known to be a crucial cytokine in the defense against pathogens, which among other responses is due to inducing epithelial repair (Lindemans et al, 2010), and by inducing antimicrobials such as REG3B and REG3g (Zheng et al, 2008). To test if inhibition of LSD1 would affect the IL-22induced antimicrobial responses, we co-treated WT organoids with a potent LSD1 inhibitor that phenocopies Lsd1 deletion (Zwiggelaar et al, 2020) and IL-22. We found that IL-22 induced STAT3 Tyr705 phosphorylation by western blot and confocal imaging, and robustly induced Reg3b and Reg3g by qPCR independently of LSD1 enzymatic activity (Fig.…”

“…In the intestinal epithelium, the importance of maintaining epigenetic marks is exemplified by the finding that loss of the polycomb repressor complex 2 (PRC2) leads to a rapid loss of various cell types including intestinal stem cells, which subsequently leads to loss of tissue integrity and mice become moribund (Chiacchiera et al, 2016;Jadhav, 2017;Koppens et al, 2016). We recently found an important role for the demethylase LSD1 in early-life intestinal epithelial development, including the differentiation of Paneth cells (Zwiggelaar et al, 2020). LSD1 can demethylate mono or dimethylated Histone 3 Lysine 4 (H3K4me1/2) that normally mark active or poised enhancers (Shi et al, 2004).…”