Polycomb Repressive Complex 2 Regulates Genes Necessary for Intestinal Microfold Cell (M Cell) Development

Abstract

Chromatin immunoprecipitation and globalQ8 run-on sequencing analysis of gut Microfold cells (M cells) showed 12 previously unknown novel transcription factors and, one of them, estrogen-related-receptor g, plays a critical role in M-cell differentiation. Lack of estrogen-relatedreceptor g showed an immature and nonfunctional M-cell phenotype. BACKGROUND & AIMS: Microfold cells (M cells) are immunosurveillance epithelial cells located in the Peyer's patches (PPs) in the intestine and are responsible for monitoring and transcytosis of antigens, microorganisms, and pathogens. Mature M cells use the receptor glycoprotein 2 (Gp2) to aid in transcytosis. Recent studies have shown transcription factors, Spi-B and Sox8 Q9 . are necessary for M-cell differentiation, but not sufficient. An exhaustive set of factors sufficient for differentiation and development of a mature Gp2þ M cell remains elusiveQ10. Our aim was to understand the role of polycomb repressive complex 2 (PRC2) as an epigenetic regulator of Mcell development. Estrogen-related-receptor g (Esrrg), identified as a PRC2-regulated gene, was studied in depth, in addition to its relationship with Spi-B and Sox8.METHODS: Comparative chromatin immunoprecipitation and global run-on sequencing analysis of mouse intestinal organoids were performed in stem condition, enterocyte conditions, and receptor activator of nuclear factor k B ligand-induced M-cell condition. Esrrg, which was identified as one of the PRC2-regulated transcription factors, was studied in wild-type mice and knocked out in intestinal organoids using Crispr Cas9 Q11. Sox8 null mice were used to study Esrrg and its relation to Sox8.RESULTS: chromatin immunoprecipitation and global run-on sequencing analysis showed 12 novel PRC2 regulated transcription factors, PRC2-regulated Esrrg is a novel M-cell-specific transcription factor acting on a receptor activator of nuclear factor kB ligand-receptor activator of nuclear factor kB-induced nuclear factor-kB pathway, upstream of Sox8, and necessary but not sufficient for a mature M-cell marker of Gp2 expression. CONCLUSIONS: PRC2 regulates a significant set of genes in M cells including Esrrg, which is critical for M-cell development and differentiation. Loss of Esrrg led to an immature M-cell phenotype lacking in Sox8 and Gp2 expression. Transcript profiling: the data have been deposited in the NCBI Gene Expression Omnibus database (GSE157629). (Cell Mol

PRC2 Regulated Atoh8 Is a Regulator of Intestinal Microfold Cell (M Cell) Differentiation

“…Previously in osteoclast, it was shown that BMP-induced Atoh8 regulated the RankL/OPG ratio indirectly via Runx2 to regulate osteoclast number and maintain bone volume in mice [20]. Our Gro-seq analysis of RankL organoids from our previous paper revealed that RankL signaling did lead to an upregulation of BMP2 and BMP6 but not Runx2 [19]. The BMP6 and BMP2 activation by RankL was confirmed by qPCR analysis (Supplementary Figure S3A).…”

“…However, other factors controlling M cell developments remain to be elucidated. For our analysis into M cell differentiation, we had previously performed a Chip-seq and Gro-seq of M cells and our data set revealed that PRC2 regulated 12 novel transcription factors (six silenced and six upregulated) in M cell development [19]. Atoh8 was one of the six transcription factors upregulated by the PRC2.…”

“…In our previous study, we sought to identify how PRC2 regulates M cell differentiation and discovered 12 transcription factors that were PRC2-regulated specifically in M cell differentiation (six upregulated and six silenced) [19]. Atoh8 turned up as one of the RankLinduced PRC2 regulated genes (log2 fold changes −3.15 RankL vs WENRC and -2.58 RankL vs ENRI) (Figure 1A).…”

“…To further understand the differentiation and development of M cells, we had previously performed Chip-seq and Gro-seq for M cells and discovered 12 transcription factors that were epigenetically regulated (six upregulated and six silenced) [19]. One of the genes upregulated in our analysis was Atoh8.…”

Immune Responses to Orally Administered Recombinant Lactococcus lactis Expressing Multi-Epitope Proteins Targeting M Cells of Foot-and-Mouth Disease Virus

“…M cells, which are epithelial cells, are localized in Peyer's patches (PPs) in the intestines, nasopharynx-related lymphoid tissue (NALT), isolated lymphoid follicles, and the appendix. These cells are responsible for the monitoring and transcytosis of antigens, microorganisms, and pathogens [18,19]. M cells play vital roles as gatekeepers of mucosal immunity, even though, in the intestinal tract, only 1 in 10 million epithelial cells is an M cell [20].…”