Project description:In our laboratory, we developed a new knock-in murine model to trace p16 cells ex vivo using EGFP as a reporter gene under the control of the endogenous promoter (exon 3) of the p16 locus from the CDKN2A gene (p16-Cre/Rosa26-mtmg). We showed previously that cells positive for p16-EGFP accumulated during natural aging. We previously reported (Grosse et al, 2020) that among this cells, liver endothelial cells and macrophages from liver and other tissues are the main population going into p16 activation. We are now analyzing the characteristics of p16High tissue resident peritoneal macrophages to understant the roll they have during adulhood and aging.
Project description:We report high-throughput profiling of gene expression from murine primary macrophages. We profile mRNA in control and endotoxin stimulated macrophages, and examine the effect of AHR ligand (SGA360) under inflammatory status. This study provides a framework for understanding transcriptional changes caused by SGA360 during activated inflammatory signaling .
Project description:The accumulation of senescent cells can drive many age-associated phenotypes and pathologies. Consequently, it has been proposed that removing senescent cells might extend lifespan. Here we generated two knock-in mouse models targeting the best-characterized marker of senescence, p16Ink4a. Using a genetic lineage tracing approach, we found that age-induced p16High senescence is a slow process that manifests around 10-12 months of age. The majority of p16High cells were vascular endothelial cells mostly in liver sinusoids (LSECs), and to lesser extent macrophages and adipocytes. In turn, continuous or acute elimination of p16High senescent cells disrupted blood–tissue barriers with subsequent liver and peri-vascular tissue fibrosis and health deterioration. Our data show that senescent LSECs are not replaced after removal and have important structural and functional roles in the aging organism. In turn, delaying senescence or replacement of senescent LSECs could represent a powerful tool in slowing down aging.
Project description:To examine the miRNA expression profile of the profibrogenic macrophages, we compared miRNA expression in untreated versus IL-4 or IL-13-treated macrophages using miRNA microarray.IL-4 and IL-13 treatment resulted in marked changes in 18 and 19 miRNAs in macrophages, respectively.(n=3, fold changes>2, P<0.05) Total RNA from human primary macrophages (n=3) treated with or without 20 ng/ml IL-4 or 20ng/ml IL-13 for 48 hr was isolated with a mirVana miRNA Isolation kit (Ambion, Foster, CA). The miRNA expression profile of the samples were examined using miRCURY LNAM-bM-^DM-" arrays (v18.0; covering 1921 human miRNAs, Exiqon ,Vedbaek, Denmark).
Project description:We used scRNA-seq analysis to investigate the expression profiles of p16high adipocyte progenitor cells compared with p16low adipocyte progenitor cells in the gonadal adipose tissue of aging mice.
Project description:To determine all ubiquitination modified proteins and corresponding ubiquitinated sites, we performed di-Gly LC/MS in mouse primary macrophages (MG132-treated).