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Single-cell transcriptomics of adult gd T-cell lineage commitment highlights its TCR-instructive nature

ABSTRACT: This SuperSeries is composed of the SubSeries listed below.

ORGANISM(S): Mus musculus

PROVIDER: GSE184545 | GEO | 2022/01/01

REPOSITORIES: GEO

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Single-cell transcriptomics of adult gd T-cell lineage commitment highlights its TCR-instructive nature II

Project description:Single cell RNA-seq study of T lymphocyte differentiation WT and LAT deficient mice in order to deciphy gd T-cell lineage commitment.

2022-01-01 | GSE184544 | GEO

Single-cell transcriptomics of adult gd T-cell lineage commitment highlights its TCR-instructive nature I

Project description:Single cell RNA-seq study of T lymphocyte differentiation WT and LAT deficient mice in order to deciphy gd T-cell lineage commitment.

2022-01-01 | GSE184483 | GEO

Single-cell transcriptomics of adult gd T-cell lineage commitment highlights its TCR-instructive nature

Project description:Single-cell transcriptomics of adult gd T-cell lineage commitment highlights its TCR-instructive nature

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Single-cell transcriptomics of adult gd T-cell lineage commitment highlights its TCR-instructive nature II

Project description:Single-cell transcriptomics of adult gd T-cell lineage commitment highlights its TCR-instructive nature II

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Single-cell transcriptomics of adult gd T-cell lineage commitment highlights its TCR-instructive nature I

Project description:Single-cell transcriptomics of adult gd T-cell lineage commitment highlights its TCR-instructive nature I

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Single-cell transcriptomics uncovers an instructive T-cell receptor role in adult γδ T-cell lineage commitment

Project description:After entering the adult thymus, bipotent T-cell progenitors give rise to αβ- or γδ-T-cells. To determine whether the γδ T-cell receptor (TCR) has an instructive role in γδ-T-cell lineage commitment or only 'confirms' a pre-established γδ-Τ cell lineage state, we exploited mice lacking expression of LAT, an adaptor required for γδ TCR signaling. Although these mice showed a T-cell development block at the CD4-CD8- double-negative (DN) 3 stage, 0.3% of their DN3 cells expressed intermediate levels of γδ-TCR (further referred to as γδint) at their surface. Single-cell transcriptomics of LAT-deficient DN3 γδint cells demonstrated no sign of commitment to the γδ-T cell lineage, apart from γδ-TCR expression. Although the lack of LAT is thought to tightly block DN3 cell development, we unexpectedly found that 25% of LAT-deficient DN3 γδint cells were actively proliferating and progressed up to the DN4 stage. However, even those cells failed to turn on the transcriptional program associated with the γδ-T-cell lineage. Therefore, the γδ-TCR-LAT signaling axis builds upon a γδ-T-cell uncommitted lineage state to fully instruct adult γδ-T-cell lineage specification.

| S-SCDT-EMBOJ-2021-110023 | biostudies-other

Single-cell transcriptomics uncovers an instructive T-cell receptor role in adult γδ T-cell lineage commitment.

Project description:After entering the adult thymus, bipotent T-cell progenitors give rise to αβ or γδ T cells. To determine whether the γδ T-cell receptor (TCR) has an instructive role in γδ T-cell lineage commitment or only "confirms" a pre-established γδ Τ-cell lineage state, we exploited mice lacking expression of LAT, an adaptor required for γδ TCR signaling. Although these mice showed a T-cell development block at the CD4- CD8- double-negative third (DN3) stage, 0.3% of their DN3 cells expressed intermediate levels of γδ TCR (further referred to as γδint ) at their surface. Single-cell transcriptomics of LAT-deficient DN3 γδint cells demonstrated no sign of commitment to the γδ T-cell lineage, apart from γδ TCR expression. Although the lack of LAT is thought to tightly block DN3 cell development, we unexpectedly found that 25% of LAT-deficient DN3 γδint cells were actively proliferating and progressed up to the DN4 stage. However, even those cells failed to turn on the transcriptional program associated with the γδ T-cell lineage. Therefore, the γδ TCR-LAT signaling axis builds upon a γδ T-cell uncommitted lineage state to fully instruct adult γδ T-cell lineage specification.

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Dynamics of lineage commitment revealed by single-cell transcriptomics of differentiating embryonic stem cells.

Project description:Gene expression heterogeneity in the pluripotent state of mouse embryonic stem cells (mESCs) has been increasingly well-characterized. In contrast, exit from pluripotency and lineage commitment have not been studied systematically at the single-cell level. Here we measure the gene expression dynamics of retinoic acid driven mESC differentiation from pluripotency to lineage commitment, using an unbiased single-cell transcriptomics approach. We find that the exit from pluripotency marks the start of a lineage transition as well as a transient phase of increased susceptibility to lineage specifying signals. Our study reveals several transcriptional signatures of this phase, including a sharp increase of gene expression variability and sequential expression of two classes of transcriptional regulators. In summary, we provide a comprehensive analysis of the exit from pluripotency and lineage commitment at the single cell level, a potential stepping stone to improved lineage manipulation through timing of differentiation cues.

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Enforcement of ??-lineage commitment by the pre-T-cell receptor in precursors with weak ??-TCR signals.

Project description:Developing thymocytes bifurcate from a bipotent precursor into ??- or ??-lineage T cells. Considering this common origin and the fact that the T-cell receptor (TCR) ?-, ?-, and ?-chains simultaneously rearrange at the double negative (DN) stage of development, the possibility exists that a given DN cell can express and transmit signals through both the pre-TCR and ??-TCR. Here, we tested this scenario by defining the differentiation outcomes and criteria for lineage choice when both TCR-? and ??-TCR are simultaneously expressed in Rag2(-/-) DN cells via retroviral transduction. Our results showed that Rag2(-/-) DN cells expressing both TCRs developed along the ??-lineage, down-regulated CD24 expression, and up-regulated CD73 expression, showed a ??-biased gene-expression profile, and produced IFN-? in response to stimulation. However, in the absence of Inhibitor of DNA-binding 3 expression and strong ??-TCR ligand, ??-expressing cells showed a lower propensity to differentiate along the ??-lineage. Importantly, differentiation along the ??-lineage was restored by pre-TCR coexpression, which induced greater down-regulation of CD24, higher levels of CD73, Nr4a2, and Rgs1, and recovery of functional competence to produce IFN-?. These results confirm a requirement for a strong ??-TCR ligand engagement to promote maturation along the ?? T-cell lineage, whereas additional signals from the pre-TCR can serve to enforce a ??-lineage choice in the case of weaker ??-TCR signals. Taken together, these findings further cement the view that the cumulative signal strength sensed by developing DN cells serves to dictate its lineage choice.

| S-EPMC3992653 | biostudies-literature

Calcium dependent CAMTA1 in adult stem cell commitment to a myocardial lineage.

Project description:The phenotype of somatic cells has recently been found to be reversible. Direct reprogramming of one cell type into another has been achieved with transduction and over expression of exogenous defined transcription factors emphasizing their role in specifying cell fate. To discover early and novel endogenous transcription factors that may have a role in adult-derived stem cell acquisition of a cardiomyocyte phenotype, mesenchymal stem cells from human and mouse bone marrow and rat liver were co-cultured with neonatal cardiomyocytes as an in vitro cardiogenic microenvironment. Cell-cell communications develop between the two cell types as early as 24 hrs in co-culture and are required for elaboration of a myocardial phenotype in the stem cells 8-16 days later. These intercellular communications are associated with novel Ca(2+) oscillations in the stem cells that are synchronous with the Ca(2+) transients in adjacent cardiomyocytes and are detected in the stem cells as early as 24-48 hrs in co-culture. Early and significant up-regulation of Ca(2+)-dependent effectors, CAMTA1 and RCAN1 ensues before a myocardial program is activated. CAMTA1 loss-of-function minimizes the activation of the cardiac gene program in the stem cells. While the expression of RCAN1 suggests involvement of the well-characterized calcineurin-NFAT pathway as a response to a Ca(2+) signal, the CAMTA1 up-regulated expression as a response to such a signal in the stem cells was unknown. Cell-cell communications between the stem cells and adjacent cardiomyocytes induce Ca(2+) signals that activate a myocardial gene program in the stem cells via a novel and early Ca(2+)-dependent intermediate, up-regulation of CAMTA1.

| S-EPMC3371086 | biostudies-literature

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