Project description:Asbestos-associated diseases remain a social burden worldwide. Our previous studies identified asbestos-induced iron-rich milieu for mesothelial cells with ceaseless macrophage ferroptosis. However, molecular mechanisms how this mutagenic milieu influences mesothelial cells have not been elucidated yet. Here, we propose a novel mechanism that extracellular vesicles (EVs) mediate asbestos-associated mutagenic factors to mesothelial cells. In a mice model of intraperitoneal crocidolite injection, mutagenic milieu highly expressed CD63, an exosomal marker. We then used a CD63-GFP labeled THP-1 macrophage model exposed to crocidolite/iron, which generated EVs under ferroptotic process. We observed that MeT-5A mesothelial cells can receive and internalize these EVs. Furthermore, we comprehensively analyzed the ferroptosis-dependent EVs (FedEVs) for transported proteins and identified ferritin heavy/light chains as major proteins. Therefore, we inferred that FedEVs transport iron from ferroptotic macrophages to mesothelial cells. RNA sequencing revealed that the mesothelial cells receiving higher amounts of the FedEVs were mitotic, especially at the S and G2/M phases, by the use of Fucci mesothelial cells. Nuclear 8-hydroxy-2`-deoxyguanosine and γ-H2AX were significantly increased in the recipient mesothelial cells after exposure to FedEVs. Collectively, we here demonstrate a novel mechanism that FedEVs act as a key mutagenic mediator by transporting iron, which contribute to asbestos-induced mesothelial carcinogenesis.
Project description:To characterize the mesothelial cell-mediated changes of gene expression in ovarian cancer cells, we performed RNA sequencing in OVCAR8 cells with and without mesothelial cell co-culture.
Project description:Characterization and Functional Analyses of Hepatic Mesothelial Cells in Mouse Liver Development. We used microarrays to detail the global programme of gene expression in mesothelial cells during mouse liver development. Mouse liver mesothelial cells were selected by a cell sorter at E12.5 and adult for RNA extraction and hybridization on Affymetrix microarrays. We obtained PCLP1+ cells and Mesothelin+ cells at each developmental stage in order to analyze expression profiles of mesothelial cells.
Project description:Characterization and Functional Analyses of Hepatic Mesothelial Cells in Mouse Liver Development. We used microarrays to detail the global programme of gene expression in mesothelial cells during mouse liver development.
Project description:RNA-sequencing analysis was carried out on ascetic fluid-isolated mesothelial cells from ovarian cancer patients compared to control human peritoneal mesothelial cells to identify a mesothelial-mesenchymal gene signature.
Project description:Bulk RNA-seq was performed on fibroblasts and mesothelial cells from 11 different healthy mouse tissues such as bone, epididymal, brown and inguinal fat pads, mesentery, omentum, pancreas, liver, lung, lymph node and spleen. This data was used to examine tissue specific gene expression programs of fibroblasts and mesothelial cells.
Project description:To analyze p57 function in mouse fetal liver, we surgically isolated mesothelial and submesothelial cells from E17 p57-/- and p57+/+ livers. Compared gene expression in E17 p57-/- and p57+/+ hepatic mesothelial and submesothelial cells.
Project description:Local factors produced in the tissue microenvironment play essential roles in promoting the ontogeny and phenotype of tissue resident macrophages (TRM). In the peritoneal cavity, large peritoneal macrophages (LPM) are the dominant TRMs that functionally mediate type 2 immunity, facilitate tissue repair of the mesothelium, and protect against peritoneal fibrosis. It is established that retinoic acid derived from the omentum induces transcription factor Gata6 expression in LPMs, which in turn regulates gene expression of factors that define peritoneal macrophages. It is still unclear whether retinoic acid is the sole local factor that regulates Gata6 expression in LPMs. Mesothelial cells line the entire peritoneal cavity and produce a protective, non-adhesive barrier against injury, at least in part by recruiting immune cells with secreted cytokines, such as M-CSF. We hypothesized that secreted factors from peritoneal mesothelial cells are also responsible for regulating LPM development including both ontogeny and function. Due to their immediate proximity to the peritoneal cavity, we propose that mesothelial cells can produce and secrete proteins into the peritoneum to maintain Gata6 expression by LPMs. To identify secreted factors that are highly and specifically expressed in mesothelial cells, we harvested primary mesothelial cells from 10-week-old C57BL/6 mice using FACS selection (CD45- PDPN+ GPM6a+). Total RNA was isolated from these cells and subjected to RNA-seq analysis after depletion of ribosomal RNA.