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Generation of Tumor Antigen-Specific iPSC-Derived Thymic Emigrants (iTE) using a Novel 3D Thymic Culture System

ABSTRACT: Generation of Tumor Antigen-Specific iPSC-Derived Thymic Emigrants (iTE) using a Novel 3D Thymic Culture System

PROVIDER: PRJNA414659 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR6184200_1.fastq.gz	Fastqsanger.gz
	SRR6184200_2.fastq.gz	Fastqsanger.gz
	SRR6184210_1.fastq.gz	Fastqsanger.gz
	SRR6184210_2.fastq.gz	Fastqsanger.gz
	SRR6184201_1.fastq.gz	Fastqsanger.gz

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Generation of Tumor Antigen-Specific iPSC-Derived Thymic Emigrants (iTE) using a Novel 3D Thymic Culture System

Project description:Induced Pluripotent Stem Cell (iPSC)-derived T cells may provide future therapies for cancer patients, but those generated by current methods, such as the OP9/DLL1 system, have shown abnormalities that pose major barriers for clinical translation. Our data indicate that these iPSC-derived CD8 single positive T cells are more similar to CD4+CD8+ double positive T cells than to mature naïve T cells, display markers of developmental arrest and exhibit characteristics of innate-like immune cells. We developed a 3D organ culture method which avoids these aberrant developmental fates, and generates a novel homogenous subset of CD8αβ+ antigen-specific T cells, iPSC-derived Thymic Emigrants (iTE). iTE exhibit phenotypic and functional similarities to naïve T cells both in vitro and in vivo, including an enormous capacity for expansion, memory formation and tumor suppression. These data illustrate the limitations of current methods and provide a blueprint for the generation of antigen-specific iTE with tremendous clinical potential.

2018-02-28 | GSE105110 | GEO

Generation of Tumor Antigen-Specific iPSC-Derived Thymic Emigrants Using a 3D Thymic Culture System.

Project description:Induced pluripotent stem cell (iPSC)-derived T cells may provide future therapies for cancer patients, but those generated by current methods, such as the OP9/DLL1 system, have shown abnormalities that pose major barriers for clinical translation. Our data indicate that these iPSC-derived CD8 single-positive T cells are more like CD4+CD8+ double-positive T cells than mature naive T cells because they display phenotypic markers of developmental arrest and an innate-like phenotype after stimulation. We developed a 3D thymic culture system to avoid these aberrant developmental fates, generating a homogeneous subset of CD8αβ+ antigen-specific T cells, designated iPSC-derived thymic emigrants (iTEs). iTEs exhibit phenotypic and functional similarities to naive T cells both in vitro and in vivo, including the capacity for expansion, memory formation, and tumor suppression. These data illustrate the limitations of current methods and provide a tool to develop the next generation of iPSC-based antigen-specific immunotherapies.

| S-EPMC5930030 | biostudies-literature

AIRE drives early life thymic interferons and changes thymic self-antigen expression [bulk RNA-seq]

Project description:AIRE drives early life thymic interferons and changes thymic self-antigen expression [bulk RNA-seq]

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Aire regulates transfer of antigen from mTEC to dendritic cells for induction of thymic tolerance

Project description:To investigate the role of Aire in thymic selection, we examined the cellular requirements for generation of ovalbumin (OVA)-specific CD4 and CD8 T cells in mice expressing OVA under the control of the rat insulin promoter (RIP). Aire-deficiency reduced the number of mature single positive (SP) OVA-specific CD4+ or CD8+ T cells in the thymus, independent of OVA expression. Importantly, it also contributed in two way to OVA-dependent negative selection depending on the T cell type. Aire-dependent negative selection of OVA-specific CD8 T cells correlated with Aire-regulated expression of OVA. By contrast, for OVA-specific CD4 T cells, Aire affected tolerance induction by a mechanism that operated independent of the level of OVA expression, controlling access of antigen presenting cells to mTEC-expressed OVA. This study supports the view that one mechanism by which Aire controls thymic negative selection is by regulating the indirect presentation of mTEC-derived antigens by thymic dendritic cells. It also indicates that mTEC can mediate tolerance by direct presentation of Aireregulated antigens to both CD4 and CD8 T cells. Total RNA was extracted from medullary thymic epithelial cells isolated from wildtype and Aire-deficient C57BL/6 mice for comparison of gene expression profiles.

2011-04-06 | E-GEOD-28393 | biostudies-arrayexpress

RNA-seq data from T cells in vitro

Project description:Comparison of athymic T cells generated in vitro with iPSC-derived recent thymic emigrant (iRTE) generated from 3D thymic culture and natural splenocytes.

2019-11-17 | GSE89989 | GEO

Gene expression profiling on 2D vs 3D iPSC colonies

Project description:Gene expression profiling on 2D vs 3D iPSC colonies

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Expression data from meduallar thymic epithelial cells and thymic or splenic B cells

Project description:Aire is a transcriptional regulator that induces promiscuous expression of thousands of tissue-restricted antigen (TRA) genes in medullary thymic epithelial cells (mTECs). While the target genes of Aire are well characterized, the transcriptional programs regulating its own expression remain elusive. We used Affymetrix microarrays to analyze the gene expression patterns of Aire expressing cells (mature mTECs and Thymic B cells) and compared them to control counterparts, namely immature mTECs, cortical Thymic epithelial cells and splenic B cells of tissue-restricted antigen (TRA) genes in medullary thymic epithelial cells (mTECs). While the target genes of Aire are well characterized, the transcriptional programs regulating its own expression remain elusive. We used Affymetrix microarrays to analyze the gene expression patterns of Aire expressing cells (mature mTECs and Thymic B cells) and compared them to control counterparts, namely immature mTECs, cortical Thymic epithelial cells and splenic B cells.

2016-12-12 | GSE89965 | GEO

Single cell RNA sequencing of thymic dendritic cells reveals molecular determinants of antigen transfer in the thymus

Project description:The presentation of self-antigens in the thymus by medullary thymic epithelial cells (mTECs) and dendritic cells (DCs) is crucial for the establishment of central tolerance. While mTECs produce and present self-antigens in an autonomous manner, DCs acquire them from mTECs by Cooperative Antigen Transfer (CAT). Although CAT represents a key process of central tolerance, molecular determinants which drive CAT are largely unknown. To reveal these determinants, we compared the transcriptomes of CAT-experienced and CAT-inexperienced DCs via scRNAseq. Specifically, we sequenced TdTOMATO+ and TdTOMATO– CD11c+ and CD11b+ cells from the thymus of Foxn1CreR26TdTOMATO mice. In this model, fluorescent TdTOMATO protein is exclusively produced by thymic epithelium, thus TdTOMATO expression in DCs (CD11c+ and CD11b+ cells) mark their previous experience of CAT. Using this approach we unraveled several candidate determinants of CAT. We focused on the gene determinant, Cldn1, encoding the tight junction protein Claudin 1, that was found to be highly expressed among the CAT-experienced type 1 lineage DCs. We performed further experiments that confirmed its role in driving CAT. In addition, this scRNAseq approach allowed us to redefine the heterogeneity and maturation states of thymic DCs.

2024-07-31 | E-MTAB-14319 | biostudies-arrayexpress

AIRE drives early life thymic interferons and changes thymic self-antigen expression [bulk RNA-seq]

Project description:Type I interferons (IFN-I) are transiently induced during infections, but were reported to be produced in the thymus at steady state. We found that developing thymocytes displayed an IFN-I signature that was mainly dependent on IFN-β. Using Ifnb1 reporter mouse strains, we observed IFN-β expression in a small population of medullary thymic epithelial cells (mTEC), which was AIRE-dependent and peaked at 2-3 weeks of age. To study the cellular response to thymic IFN, we used a Mx1gfp reporter mouse strain and found that the IFN response in T cells and myeloid cells was abrogated by IFNaR deficiency, whereas the response in B cells and dendritic cells was dependent on both IFNaR and IFNlR. Furthermore, single-cell RNAseq analysis revealed dramatic transcriptional changes in all thymic APCs in IFNaR/IFNlR deficient mice, and a loss of CCR7+ cDC1 cells. Together, these results show that steady state IFN-I and IFN-III signaling drives a gene-expression program in thymic APCs that shapes the thymic microenvironment.

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LC-MSMS of the MHC-I immunopeptidome of murine thymic epithelial cells

Project description:Antigen presentation by cortical and medullary thymic epithelial cells (cTEC and mTEC) ensures the formation of a self-restricted and self-tolerant T cell repertoire, respectively. As such, a broad diversity of self-antigens needs to be presented by mTEC to induce T cell’s self-tolerance. Even though the expression and antigen presentation of protein coding genes in mTEC has been abundantly described, little is known of the implication of allegedly noncoding regions of the genome to tolerance induction. In this study, we focused on transposable elements (TE), which have been shown to be highly expressed by mTEC.

2024-04-17 | PXD042241 | Pride

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