Integration of phospho-signaling and transcriptomics in single cells reveals distinct Th17 cell fates

Summary: 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, an innovative platform that integra...

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Main Authors: Seth D. Fortmann, Awalpreet S. Chadha, Blake F. Frey, Asif Elahi, Vidya Sagar Hanumanthu, Shanrun Liu, Andrew Goldsborough, P. Brent Ferrell, Jr., Maria B. Grant, Casey T. Weaver, Robert S. Welner
Format: Article
Language:English
Published: Elsevier 2025-08-01
Series:Cell Reports
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Online Access:http://www.sciencedirect.com/science/article/pii/S2211124725007776
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Summary:Summary: 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, an innovative platform that integrates scRNA-seq and intracellular cellular indexing of transcriptomes and epitopes by sequencing 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 interleukin-2 (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.
ISSN:2211-1247