Mitochondrial perturbation in the intestine causes microbiota-dependent injury and gene signatures discriminative of inflammatory disease
Ontology highlight
ABSTRACT: Single-cell RNA-sequencing (scRNA-seq) data was generated from colonic tissue from Hsp60Δ/ΔIEC and Hsp60fl/fl mice at day 8 enriched in intestinal epithelial cells (IECs).
Project description:In vitro differentiation of human pluripotent stem cells into functional retinal pigment epithelial (RPE) cells for cell based reparative therapy of age-related macular degeneration (AMD). In the present study, we identify CD140b, CD56, GD2 and CD184 as central cell surface markers to evaluate hPSC-RPE differentiation efficiency, as well as a potential tool for the enrichment of hPSC-RPE during and after differentiation. Using these markers together with single cell RNA sequencing to evaluate the differentiation process, we have established an efficient xeno-free and defined monolayer differentiation methodology where culture on supportive human recombinant laminin eliminates the need for manual selection, allowing large-scale production of pure hPSC-RPE.
Project description:Single cell transcriptome showed TGF-β expression is confined to PD-L1+ endothelium and M2 /lipofibroblast-like cells. Hence, superior effects of BA could be attributed to its ability to trap TGF-β to relevant PD-L1+ compartments. Mice were irradiated (photon) and one group of mice was treated with Bintrafusp. Lungs were subsequently dissociated and processed with the 10x genomics 3 prime v2 kit.
Project description:B-cell lymphoma/leukemia (BCL) 11B represents a major regulator of vital processes in post-thymic lymphocytes as well as of lymphocyte differentiation by blocking NK cell and promoting T cell development and is thus considered a \\"guardian of T cell fate\\". Interestingly, on the one hand, NK cell maturation correlates with increased expression of BCL11B, on the other hand, native or induced T cell populations characterized by multiple NK cell features showed reduced BCL11B expression. Both NK cells or T cells display unique and desired features for cellular therapies. NK cells are able to efficiently lyse cancerous or virally infected cells but can only be poorly expanded in vitro. In contrast, T cell cytotoxic activity is limited by T cell receptor (TCR) but in vitro expansion is easily achievable. We show that from buffy coats of healthy donors isolated and CRISPR/Cas9-mediated BCL11B knock-out CD8+ T cells stimulated with IL-15 acquired remarkable innate characteristics and combine spontaneous and antibody-dependent NK cell cytotoxicity with T cell capability for rapid in vitro expansion. The observed features of the induced innate CD8+ (iiT8) cells make them an interesting and promising tool for cellular therapy.
Project description:The Ets transcription factor, ERG, plays a central role in definitive hematopoiesis and its overexpression in acute myeloid leukemia is associated with a stem cell signature and bad prognosis. However, little is known about the underlying mechanism by which ERG causes leukemia. Therefore we sought to identify ERG targets that participate in development of leukemia by integration of expression arrays and Chromatin immunoprecipitation. Bone marrow was collected from three transgenic ERG mice harboring a myeloid leukemia and twelve wild type mice. Nine wild type bone marrow samples were lineage depleted and pooled into 3 samples (each one consisting of three lineage negative wild type bone marrows). The resulting 9 samples were used for RNA extraction and hybridization on affymetrix microarrays.
Project description:The naked mole-rat (NMR; Heterocephalus glaber) has recently gained considerable attention in the scientific community for its unique potential to unveil novel insights in the fields of medicine, biochemistry, and evolution. NMRs exhibit unique adaptations that include protracted fertility, cancer resistance, eusociality, and anoxia. This suite of adaptations is not found in other rodent species, suggesting that interrogating conserved and accelerated regions in the NMR genome will find regions of the NMR genome fundamental to their unique adaptations. However, the current NMR genome assembly has limits that make studying structural variations, heterozygosity, and non-coding adaptations challenging. We present a complete diploid naked-mole rat genome assembly by integrating long-read and 10X-linked read genome sequencing of a male NMR and its parents, and Hi-C sequencing in the NMR hypothalamus (N=2). Reads were identified as maternal, paternal or ambiguous (TrioCanu). We then polished genomes with Flye, Racon and Medaka. Assemblies were then scaffolded using the following tools in order: Scaff10X, Salsa2, 3d-DNA, Minimap2-alignment between assemblies, and the Juicebox Assembly Tools. We then subjected the assemblies to another round of polishing, including short-read polishing with Freebayes. We assembled the NMR mitochondrial genome with mitoVGP. Y chromosome contigs were identified by aligning male and female 10X linked reads to the paternal genome and finding male-biased contigs not present in the maternal genome. Contigs were assembled with publicly available male NMR Fibroblast Hi-C-seq data (SRR820318). Both assemblies have their sex chromosome haplotypes merged so that both assemblies have a high-quality X and Y chromosome. Finally, assemblies were evaluated with Quast, BUSCO, and Merqury, which all reported the base-pair quality and contiguity of both assemblies as high-quality. The assembly will next be annotated by Ensembl using public RNA-seq data from multiple tissues (SRP061363). Together, this assembly will provide a high-quality resource to the NMR and comparative genomics communities.
Project description:Aim: To assess the gene expression profiles of peripheral blood mononuclear cells obtained from palindromic rheumatism patients during flares and between flares. Samples were paired so we could assess the differences in gene expression profiles per a given patient during a flare and between flares. Method: PBMCs were extracted from whole blood and run on Illumina HT-12 v4 beadchips. The raw data from Illumina’s iScan software were read into R (version 3.4.4) and processed (background corrected, normalised and summarised) using the beadarray package (version 2.34.0). Linear models made using Limma (version 3.34.9) were used to assess differential expression of genes (DEGs).
Project description:NCAM1+ subpopulation of adult human kidney epithelial cells (hKEpC) can be specifically activated in vitro to stem-like cells that de-differentiate and recapitulate embryogenesis. We used microarrays to detail the global programme of gene expression in the NCAM1+ sub-population in comparison to NCAM1-.
Project description:NCAM1+ subpopulation of fetal human kidneyl cells are nephron progenitors We used microarrays to detail the global programme of gene expression in the NCAM1+ sub-population in comparison to total culture.
Project description:Over 250 million people suffer from schistosomiasis, a tropical disease caused by parasitic flatworms known as schistosomes. To unravel the transcriptional signatures for the larval stage of this parasite, we perform single-cell RNA sequencing on two-day old schistosomula. We described 15 populations and spatially validated our results using FISH.
Project description:Gene expression profiles of individual bone marrow cells were acquired by Drop-Seq. Subsets of bone marrow cells were isolated using magnetic cell sorting to enrich for putative skeletal stem cells.