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Integration of Phospho-Signaling and Transcriptomics in Single Cells Reveals Distinct Th17 Cell Fates

ABSTRACT: Single-cell RNA sequencing (scRNA-seq) provides the resolution and scale necessary to identify transcriptional programs but fails to capture post-transcriptional information critical to decipher signaling networks and cellular states. We present Vivo-seq, a novel platform that integrates single-cell RNA-seq and intracellular CITE-seq following cellular fixation with a deep eutectic solvent, which preserves multiple domains of biological information beyond RNA transcripts alone. Vivo-seq enables simultaneous capture of both transcriptional and phospho-signaling states in single cells. Applying this platform to developing T-helper 17 (Th17) cells, we find that simultaneous phosphorylation of ERK1/2 and c-FOS leads to maximal IL-2 and IL-17 production. Furthermore, we show that early IL-2 production imprints developing Th17 cells for enhanced maintenance or cytokine-dependent transdifferentiation during subsequent antigenic stimulation. By integrating transcriptional and phospho-signaling information at single-cell resolution, we identify a hyperactivated Th17 cellular state associated with early IL-2 production that has downstream consequences on functional plasticity.

ORGANISM(S): Mus musculus

PROVIDER: GSE297075 | GEO | 2025/07/12

REPOSITORIES: GEO

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CIP2A negatively regulates Th17 differentiation

Project description:Interleukin 17 (IL-17) producing T helper 17 (Th17) cells are critical drivers of pathogenesis in a variety of autoimmune and inflammatory diseases. Strategies to mitigate excessive Th17 response thus remain an attractive target for immunotherapies. Here we report that Cancerous Inhibitor of Protein Phosphatase 2A (CIP2A) regulates IL-17 production by Th17 cells in human and mouse. Using CIP2A knock-out (KO) mice and siRNA-mediated CIP2A silencing in human primary CD4+ T cells, we demonstrated that CIP2A silencing results in a significant increase in IL-17 production. Interestingly, CIP2A deficient Th17 cells were characterized by increased strength and duration of STAT3 (Y705) phosphorylation. Genome-wide gene expression profile as well as the p-STAT3 (Y705) interactome of CIP2A deficient Th17 cells identified that CIP2A regulates the strength of the interaction between Acylglycerol kinase (AGK) and STAT3, and thereby, modulates STAT3 phosphorylation as well as expression of IL-17 in Th17 cells. Our results uncover the physiological function of CIP2A in Th17 cells and provides new opportunities for therapeutic intervention in Th17 cell mediated diseases.

2020-05-26 | PXD010612 | Pride

Single cell transcriptomic studies of mouse colonic lamina propria cells from wildtype or RORgtS182A mutant mice

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Gene array for identifying molecules involved in IL-10 regulation during Th17 polarization

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