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Dynamic interplay between IL1 and WNT pathways in regulating dermal adipocyte lineage cells during skin development and wound regeneration

ABSTRACT: In brief, skin resident cells were isolated from dorsal skin of mice and 10x Genomics single cell RNAseq was applied to identify distinct cell populations. Low quality cells and outliers were discarded, and only ～24,409 viable cells were used for downstream analysis. Unsupervised clustering and gene expression were visualized with the Seurat 2.0 on R studio, and assignment of cell clusters was based on expression of validated marker genes. Subsequent in-depth analysis was assisted by GENE DENOVO Inc (Guangzhou, China). Overall, we found that a specific dermal fibroblast population, with high DPP4, Sca1 and WNT2 expression, has robust capacity for adipocyte differentiation and acts key role in maintaining dermis homeostasis.

ORGANISM(S): Mus musculus

PROVIDER: GSE189241 | GEO | 2023/06/10

REPOSITORIES: GEO

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Dynamic interplay between IL1 and WNT pathways in regulating dermal adipocyte lineage cells during skin development and wound regeneration

Project description:Cells were isolated from wounded skin tissue and administered into single cell RNA transcriptome sequencing to deconstruct cell hierarchy of dermal fibroblast population and underlying regulating mechanisms. In brief, 7~9 weeks old male C57BL/6 mice were anesthetized and one full-thickness wound (1.0 cm x 0.5 cm in size) was generated on shaved dorsal skin. Animals were sacrificed at indicated intervals after and wound bed tissues were collected for cell isolation and subsequent 10x Genomics single cell RNAseq. Sequencing was performed on Illumina Novaseq6000 platform (Illumina). The generated raw gene expression matrix was converted into Seurat objects using the R package Seurat v3.0. After low quality cells and outliers were discarded, 10233 cells from non-lesional skin, 7807 cells from wound beds at post-wounding day 2 (PWD2), and 4925 cells from wound tissue at PWD7 were used for downstream analysis. Next, unsupervised clustering and gene expression were visualized with the Seurat 3.0 on R studio, and assignment and identification of cell clusters was based on the expression of validated marker genes. Subsequent in-depth analysis was assisted by GENE DENOVO Inc (Guangzhou, China). Together, our study has identified a specific dermal fibroblast population with high DPP4, Sca1 and WNT2 expression, which is committed into adipocyte upon wounding, and define the differentiation trajectory of this adipocyte precursor during skin regeneration.

2023-06-10 | GSE196800 | GEO

Deciphering the age-dependent changes of pulmonary fibroblasts in mice by single-cell transcriptomics

Project description:In brief, lung resident cells were isolated from lung of mice and 10x Genomics single cell RNAseq was applied to identify distinct cell populations. Low quality cells and outliers were discarded, and only ～22,826 viable cells were used for downstream analysis. These included ~5,237 cells – NB(10days mice); ~4,804 cells – Adult-M(2months mice); ~6,561cells-SD-10M (10month mice) ; ~6,224cells-SD-1d5Y (18month mice). Unsupervised clustering and gene expression were visualized with the Seurat 2.0 on R studio, and assignment of cell clusters was based on expression of validated marker genes. Subsequent in-depth analysis was assisted by GENE DENOVO Inc (Guangzhou, China). Overall, we found that a specific lung fibroblast population, We present a single-cell transcriptomic atlas encompassing lung tissues derived from mice at various stages of aging, uncovering substantial shifts in fibroblast differentiation and immune regulation throughout the aging process.

2023-11-21 | GSE237757 | GEO

Identification of a mesenchymal progenitor cell hierarchy in adipose tissue (Bulk RNA-seq)

Project description:The healthy growth of adipose tissue depends on the capacity of progenitor cells to undergo denovo adipogenesis. However, the cellular hierarchy and mechanisms governing adipocyteprogenitor differentiation are incompletely understood. Here, we identify a lineage hierarchy25 consisting of distinct mesenchymal cell types present in mouse and human adipose tissue. Cellsmarked by Dpp4 expression are highly proliferative, multipotent progenitors that give rise toIcam1+ committed pre-adipocytes and a related adipogenic population marked by Clec11a andCd142 expression. TGFβ maintains DPP4+ cell identity and inhibits adipogenic commitment ofDPP4+ and CD142+ cells. Intriguingly, DPP4+ progenitors reside in the reticular interstitium that30 envelope many organs including adipose depots. Altogether, this study defines the adipose lineagehierarchy and identifies a new anatomical niche for multipotent mesenchymal progenitors.

2019-05-01 | GSE128891 | GEO

Identification of a mesenchymal progenitor cell hierarchy in adipose tissue (scRNA-seq)

2019-05-01 | GSE128890 | GEO

Cancer associated fibroblast-derived WNT2 increases tumor angiogenesis in colon cancer

Project description:WNT2 is important for placenta vascularization and acts as a pro-angiogenic factor for liver and other endothelial cells (ECs). WNT2 induction has been shown in many carcinomas and is associated with tumor progression. In colorectal cancer (CRC) WNT2 is selectively elevated in cancer associated fibroblasts (CAFs), leading to increased invasion and metastasis. However, if there is a role for WNT2 in colon cancer angiogenesis has not been addressed so far. Here, we demonstrate that WNT2 enhances EC migration and invasion, while it induces ß catenin dependent signaling in only a small subset of HUVECs. We show that siRNA-mediated knockdown of WNT2 in CAFs reduced the growth of vessel-like structures significantly in a co-culture assay, while the overexpression of WNT2 in skin fibroblasts otherwise being devoid of WNT2 led to increased angiogenesis in vitro. In a xenograft model, overexpression of WNT2 in HCT116 led to enhanced tumor volume and vessel density. Moreover, WNT2 expression correlates with vessel markers in human CRC. Secretome profiling of CAFs revealed that proteins related to angiogenesis and extracellular matrix (ECM) remodeling are regulated by WNT2. Thus, stroma-derived WNT2 positively affects angiogenesis in CRC by shifting the balance towards pro-angiogenic signals.

2020-05-26 | PXD014019 | Pride

Repurposing DPP4-Inhibition to Improve Hair Follicle Activation and Regeneration

Project description:Skin-injury and several diseases elicit fibrosis and induce hair follicle (HF)-growth arrest and loss. Resulting alopecia and disfiguration represent a severe burden for patients both physically and psychologically. Reduction of pro-fibrotic factors such as DPP4 might be a strategy to tackle this issue. We demonstrate DPP4-overrepresentation in settings with HF-growth arrest (telogen), HF-loss and non-regenerative wound areas in mice skin and human scalp. Topical DPP4-inhibition (DPP4i) with FDA/EMA-approved Sitagliptin (Sit) on preclinical models of murine HF-activation/regeneration results in accelerated anagen-progress, while treatment of wounds with Sit results in reduced expression of fibrosis markers, increased induction of anagen around wounds, and HF-regeneration in the wound center. These effects are associated with higher expression of Wnt-target Lef1, known to be required for HF-anagen (HF-activation)/regeneration. Sit-treatment decreases pro-fibrotic signaling in the skin, induce a differentiation trajectory of HF-cells, and activate Wnt-targets related to HF-activation/growth but not those supporting fibrosis. Taken together, our study demonstrates a role for DPP4 in HF biology and shows how DPP4i, currently used as oral medication to treat diabetes, could be repurposed into a topical treatment agent to potentially reverse HF-loss in alopecia and after injury.

2023-05-02 | GSE230514 | GEO

Identification of Wnt2 targets in dendritic cell

Project description:Purpose: Next-generation sequencing (NGS) has revolutionized systems-based analysis of cellular pathways. The goals of this study are to identify the key moleculars or pathways which are changed in dendritic cells undergoing the treatment of CAF-secreted Wnt2. Methods: mRNA profiles of mouse dendritic cells (mDCs) with treatment of Wnt2, CAF culture medium (CAF.CM), or CAF culture medium + Wnt2 Ab(CAF.CM+Wnt2 Ab), were generated by deep sequencing, in triplicate, using Illumina. The sequence reads that passed quality filters were analyzed at the transcript isoform level with two methods: Burrows–Wheeler Aligner (BWA) followed by ANOVA (ANOVA) and TopHat followed by Cufflinks. The overlap changes of mRNA profiles in Wnt2-treated mDCs and CAF.CM-treated mDCs compared with control mDCs, revealed the possible changed cellular pathways induced by CAF-secreted Wnt2. The analysis of the mRNA profiles which were changed after the blocking of Wnt2 activities in CAF.CM would be used to comfirmed the changed cellular pathways induced by CAF-secreted Wnt2. Results: Using an optimized data analysis workflow, we found that 768 transcripts and 2068 transcripts were significantly changed in Wnt2-treated DCs and CAF.CM-treated respectively compared with untreated DCs. And 393 changed transcripts overlapped in Wnt2-treated DCs and CAF.CM-treated compared with untreated DCs. Moreover, treatment of CAF.CM with Wnt2 Ab induced differential expression of 497 genes. Conclusions: Our study represents the first detailed analysis of Wnt2-induced changes of DC transcriptomes, with biologic replicates, generated by RNA-seq technology. The optimized data analysis workflows reported here should provide a framework for comparative investigations of Wnt2- or CAF.CM- induecd expression profiles.

2022-01-26 | GSE154420 | GEO

Changes in gene expression in primary human white preadipocytes in response to knockdown of DPP4

Project description:Transcriptional profiling of primary human white preadipocytes after infection with lentiviral vector either containing shRNA directed against dipeptidypeptidase 4 (DPP4) or unspecific shRNA as negative control Treatment of cells with DPP4 shRNA or control shRNA was done in 4 biological replicates, the first sample of DPP4 KD was hybridized against the first sh-control sample, the second DPP4 smple against the second control sample and so on

2015-11-24 | E-GEOD-75328 | biostudies-arrayexpress

Hepatocyte-derived DPP4 regulates portal GLP-1 bioactivity, glucose production and its absence alters liver disease progression (F480+ CELLS)

Project description:Elevated circulating dipeptidyl-peptidase 4 (DPP4) is a biomarker for liver disease, but its involvement in gluconeogenesis and metabolic associated fatty liver disease (MAFLD) progression remains unclear. Here we identified that DPP4 in hepatocytes but not Tie2+ endothelial cells regulates the local bioactivity of incretin hormones and gluconeogenesis. However, the complete absence of DPP4 (Dpp4-/-) in aged mice with metabolic syndrome accelerates liver fibrosis without altering dyslipidemia and steatosis. Analysis of transcripts from the livers of Dpp4-/- mice displayed enrichment for inflammasome, p53, and senescence programs compared to littermate controls. High-fat high-cholesterol (HFHC)-feeding decreased Dpp4 expression in F4/80+ cells, with only minor changes in immune signaling.

2023-01-31 | GSE218766 | GEO

Hepatocyte-derived DPP4 regulates portal GLP-1 bioactivity, glucose production and its absence alters liver disease progression (LIVER)

2023-01-31 | GSE218765 | GEO

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