Project description:The homeostatic control mechanism of dendritic cells (DCs), including pDCs, cDC1s and cDC2s, is not fully elucidated. Transcriptome profiling of wildtype and TRIM33 conditional knockout mice revealed a key role of TRIM33 in maintaining the homeostasis of all DC subsets.
Project description:The development of dendritic cells (DCs) is regulated by complex transcriptional networks. DCs originate from the multipotent progenitors (MPPs) in the bone marrow, which could further give rise to common lymphoid progenitors (CLPs) and common dendritic cell progenitors (CDPs). Whereas CDPs, which could be further divided into CD115+ and CD115- populations, give rise to both conventional (cDC) and plasmacytoid DCs (pDCs), CLP is an addtional source of pDCs. Transcriptome profiling of wild type and Trim33-/- MPPs, CD115- CDPs, and CLPs revealed a critical role of TRIM33 in the development of DCs.
Project description:Transcriptional control of dendritic cell (DC) homeostasis has not been fully understood. Genome-wide binding analysis of TRIM33 in dendritic cells revealed a critical role in gene regulation for DC differentiation.
Project description:Transcriptional control of dendritic cell (DC) development has not been fully understood. TRIM33, a transcription co-factor was found to be crucial for transcription regulation during the development of DCs. Genome wide binding site analysis with CUT&Tag revealed co-localization of TRIM33 with CDK9 and Serine 2 phosphorylated RNA polymerase (S2 Pol II) in the common dendritic cell progenitors (CDPs).
Project description:Upon toll-like receptor (TLR) ligation, dendritic cells (DCs) undergo an activation process referred to as maturation. Over the course of maturation, global DC gene expression is severely altered, resulting in phenotypic changes including DC morphology, migration, immune stimulatory and inhibitory receptor expression, cytokine production, etc. We performed microarray analysis to investigate changes in gene expression patterns between wild type and MK2-deleted murine splenic DCs over time (0 - 24 hours) following TLR stimulation by LPS.
Project description:We analyzed the transcriptome differences of wild-type and ArhGEF1-deficient (Arhgef1-/-) type-2 conventional dendritic cells (cDC2s) in spleen. Wild-type or ArhGEF1-deficient (Arhgef1-/-) bone marrow cells were transferred into lethal irradiated CD45.1 mice. 8 weeks later, splenic cDC2s were purified and prepared for RNA-seq analysis. The gene expression profiles of CD97-, Gα13- and ArhGEF1-deficient cDC2s were highly similar, providing evidence that these molecules function in the same pathway.
Project description:The development of all dendritic cells (DCs) including antigen-presenting conventional DCs (cDCs) and cytokine-producing plasmacytoid DCs (pDCs) is controlled by the growth factor Flt3 ligand (Flt3L) and its receptor Flt3. We genetically dissected this common Flt3L-driven pathway of DCs differentiation using CRISPR/Cas9-based dropout screening in a Flt3L dependent progenitor cell line. Genome-wide screening identified multiple regulators of DC differentiation including the glycosylphosphatidylinositol transamidase complex and arginine methyltransferase Carm1, whose role was confirmed in vivo. It also showed that negative regulators of mTOR signaling, including the subunits of TSC and GATOR1 complexes, restricted progenitor growth but enabled DC differentiation, demonstrating that a TSC- and GATOR1-mediated shutdown of mTOR is required for the process. An orthogonal screen focused on transcriptional regulators showed that Trim33 (TIF-1g) is required for DC differentiation. Conditional targeting in vivo revealed an essential role of Trim33 in the development of all DCs, but not of monocytes or granulocytes. In particular, inducible deletion of Trim33 caused a rapid loss of all DC progenitors, pDCs and the cross-presenting cDC1 subset. The loss of Trim33 from DC progenitors caused spontaneous interferon and inflammatory response, aberrant induction of macrophage-specific genes, and failure to induce DC differentiation program. Collectively, these data elucidate common mechanisms that control Flt3L-driven differentiation of the entire DC lineage, and identify Trim33 as its essential regulator.
Project description:The homeostatic control of dendritic cells (DCs) is critical for appropriate immune responses, yet the regulatory mechanisms are not fully elucidated. Genome-wide mapping of TRIM33 binding sites in primary DCs revealed a role of this molecule in DC transcription regulation, and therefore homeostasis.
Project description:The transcriptome of splenic classical dendritic cells (cDCs) (CD3- CD19- CD45+ MHC-II+ CD11c+) was compared in Wild Type (WT) mice, Germ Free (GF) mice and mice deficient for the Ifnar1 gene (IFNAR). Cells were isolated and sorted by flow cytometry to high purity. The cells were stimulated ex vivo with PBS or LPS. The results show remarkable similarities between the transcriptome of splenic cDCs in GF and IFNAR mice.