Gene expression profile at single cell level of immune cells infiltrating iPS cell-derived teratoma
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ABSTRACT:
PROVIDER: PRJNA1172714 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR30986082.fastq.gz | Fastqsanger.gz | |||
| SRR30986083.fastq.gz | Fastqsanger.gz | |||
| SRR30986084.fastq.gz | Fastqsanger.gz | |||
| SRR30986085.fastq.gz | Fastqsanger.gz | |||
| SRR30986086.fastq.gz | Fastqsanger.gz |
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Project description:Currently, most cell or tissue transplantations using induced pluripotent stem cells (iPSCs) are anticipated to involve allogeneic iPSCs. However, the immunological properties of iPSCs in an allogeneic setting are not well understood. We previously established a mouse transplantation model of MHC-compatible/minor antigen-mismatched combinations, assuming a hypoimmunogenic iPSC-setting. Here, we found that iPSCs subcutaneously inoculated into MHC-compatible allogeneic host mice resisted rejection and formed teratomas without immunosuppressant administration. Notably, when skin grafts were transplanted onto hosts more than 40 days after the initial iPSCs inoculation, only the skin of the same strain as the initial iPSCs was engrafted. Therefore, donor-specific immune tolerance was induced by a single iPSC inoculation. Diverse analyses, including single-cell RNA-sequencing after transplantation, revealed an increase in regulatory T cell (Treg) population, particularly CD25+ CD103+ effector Tregs within the teratoma and skin grafts. The removal of Tregs or CD25+ cells suppressed the increase in effector Tregs and disrupted graft acceptance, indicating the importance of these cells in the establishment of immune tolerance. Within the teratoma, we observed an increase in TGF-β2 levels, suggesting an association with the increase in effector Tregs. Our results provide important insights for future applications of allogeneic iPSC-based cell or tissue transplantation.
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Project description:Human induced pluripotent stem (iPS) cells are capable of differentiating into derivatives of the three embryonic germ layers both in vitro and in vivo. To date the the molecular differences between teratoma-forming cells and non-teratoma-forming cells has not been analysed. A cell line, B1, bears typical ES cell-like morphology, expression of pluripotency-associated genes, and in vitro pluripotency capacity, but fails to form teratomas after subcutaneously injected into immune-deficient mice based on histological analysis. Besides histological analysis, we characterized the tumors derived from line B1, and teratomas derived from bona fida iPS and ES (line H1) cells respectively, using microarray-based gene expression analysis. The expression levels of pluripotency-associated markers in B1 cells were comparable to that in iPS and ES cells, while the complexity of tissue expression commitment was decreased upon spontaneous differentiation of B1 cells as compared to iPS and ES cells. Total RNA obtained from HFF1 (human foreskin fibroblast) cells, line B1, iPS-A4, iPS-B4 and ES (line H1) cells, and their derived tumors in immune-deficient mice.
2012-04-29 | E-GEOD-31970 | biostudies-arrayexpress
Project description:Human induced pluripotent stem (iPS) cells are capable of differentiating into derivatives of the three embryonic germ layers both in vitro and in vivo. To date the the molecular differences between teratoma-forming cells and non-teratoma-forming cells has not been analysed. A cell line, B1, bears typical ES cell-like morphology, expression of pluripotency-associated genes, and in vitro pluripotency capacity, but fails to form teratomas after subcutaneously injected into immune-deficient mice based on histological analysis. Besides histological analysis, we characterized the tumors derived from line B1, and teratomas derived from bona fida iPS and ES (line H1) cells respectively, using microarray-based gene expression analysis. The expression levels of pluripotency-associated markers in B1 cells were comparable to that in iPS and ES cells, while the complexity of tissue expression commitment was decreased upon spontaneous differentiation of B1 cells as compared to iPS and ES cells.
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Project description:In germ cell tumors (GCT), a growing mature teratoma during or after chemotherapy with decreasing tumor markers is defined as ´growing teratoma syndrome` (GTS) according to its first describer Logothetis et al. in 1982. Due to the small number of available cases worldwide, not much is known about this continuously growing tumor and its pathogenesis. Especially in cases with extensive and surgical uncontrollable tumor mass, specific therapeutic options and biomarkers early indicating presence of GTS are still lacking. In this study, GTS was stratified into a slow (< 0.5 cm / month), medium (0.5 – 1.5 cm / month) and rapid (> 1.5 cm / month) group based on the tumor growth rate. We analyzed the secretome of 3 GTS samples of each subgroup and 3 teratomas. The secreted proteins were isolated from ex vivo cultivated tissues and analyzed by liquid-chromatography coupled to mass spectrometry (LS-MS).
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Project description:Gene expression profile at single cell level of total tumor-infiltrating CD8+ T cells
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Project description:expression analysis of teratoma, grown from mouse embryonic stem cells that are Evi/Wls knockout cells and comparison with wildtype (control) tumors
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Project description:Three parthenogenetic induced pluripotent stem cell (PgHiPSCs) lines were generated from each of the ovarian teratoma cell lines (two distinct individuals). Two normal iPS cell lines were generated from normal fibroblasts. Three biological replicates of normal embryonic stem cells (H9, HESCs) were perfomed. We used microarrays to study the gene expression profiles of the PgHiPSCs, and compared the expression of genes to both embryonic and induced pluripotent stem cell, to identify paternally expressed genes that are down-regulated in the PgHiPSC lines. All parthenogenetic and normal iPS cell lines, were tested for pluripotency assays (inclusing, morphology, immuno stanings and qRT-PCR for known pluripotency markers, differentiation capacity in vivo and in vitro)
2011-05-31 | E-GEOD-27362 | biostudies-arrayexpress
Project description:Studying early human developmental processes requires access to a model system that is easy to perturb, either genetically or chemically, genetically tractable and recapitulates key features of organogenesis. We propose that the teratoma, a recognized standardused for for validating pluripotency in stem cells, could be a promising platform for modeling human development. Teratomas differentiate to all germ layers, have regions of complex tissue-like organization with internal vasculature, and are relatively easy to generate and thus accessible, especially in comparison to human fetal tissue. Teratomas, however, have not been characterized rigorously to determine their suitability to model human developmental processes the diversity and identities of the cell types present, and their level of maturity in comparison to human developmental cell types. We thus systematically analyzed, perturbed, and engineered human pluripotent stem cell (PSC)-derived teratomas. Usingused histology, RNA in situ hybridization, and single cell RNA-seq (scRNA-seq) of 179,632 cells across 23 teratomas generated from 4 cell lines, we observed to find that teratomas reproducibly contain approximatelyat least 20 cell types across all three germ layers, and the cell type heterogeneity between teratomas was comparable to that found in organoid systems. We found that a significant fraction of injected PSCs engrafted, suggestingused lentiviral barcoding to determine there is minimalno bottlenecking during teratoma formation. Additionally, we mapped our teratoma data to published human fetal data and showed that the teratoma gut and brain cell types correspond well to existing fetal gut and brain scRNA-seq datasetscells. We then performed a CRISPR-Cas9 knockout screen in teratomas targeting key genes known to be embryonic lethal, and found that TWIST1, RUNX1, ASCL1, CDX2, and KLF6 knockouts resulted in assessed shifts in lineage abundance. using scRNA-seq. We found that TWIST1, RUNX1, ASCL1, CDX2, and KLF6 knockouts resulted in reproducible shifts in lineage abundance consistent with known biology. All of these These knockouts had effects on cell types from multiple germ layers, showing that the teratoma can serve as a platform to studymodel all major human lineages of human development simultaneously. Additionally, we ran a CRISPR-Cas9 screen targetingWe also knocked out the genes underlying Rett, Pitt-Hopkins, and L1 Syndromes and identified cell type specific shifts in gene expression. Finally, we engineered a novel miRNA-regulated suicide gene circuit that enriches for tissues in the teratoma that express a specific miRNA that selectively depletes cells that do not express a tissue-specific endogenous miRNA, allowing us to enrich for desired lineages in the teratoma, which we demonstrated using miR-124 to enrich for neuro-ectoderm. Specifically, we used miR-124 to successfully enrich for neuro-ectoderm. Taken together, we believe the teratoma is a promising platform for modeling multi-lineage human development, functional genetic screening, and cellular engineering
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Project description:Three parthenogenetic induced pluripotent stem cell (PgHiPSCs) lines were generated from each of the ovarian teratoma cell lines (two distinct individuals). Two normal iPS cell lines were generated from normal fibroblasts. Three biological replicates of normal embryonic stem cells (H9, HESCs) were perfomed. We used microarrays to study the gene expression profiles of the PgHiPSCs, and compared the expression of genes to both embryonic and induced pluripotent stem cell, to identify paternally expressed genes that are down-regulated in the PgHiPSC lines.
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Project description:MicroRNA sequencing of slow and rapid growing teratoma. Teratoma without a growth trend were included as controls. In total, 9 samples were analyzed for their expressed microRNAs by sequencing. YST and EC tissues were identified from the biobank of the Department of Urology (University Hospital Düsseldorf). Total RNA has been isolated and subsequently microRNA sequencing has been performed.
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