{"doi":"10.1016/j.celrep.2025.116006","title":"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, 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.","journal":"Cell Reports","year":2025,"id":545382,"datarank":0.0,"base_score":0.0,"endowment":0.0,"self_citation_contribution":0.0,"citation_network_contribution":0.0,"self_endowment_contribution":0.0,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":1,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.8847,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2025-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":420335,"name":"Awalpreet S. Chadha","orcid":"0000-0002-8072-2243","position":1,"is_corresponding":false},{"id":249177,"name":"Blake Frey","orcid":"0000-0003-3003-6904","position":2,"is_corresponding":false},{"id":1068067,"name":"Asif Elahi","orcid":null,"position":3,"is_corresponding":false},{"id":259111,"name":"Vidya Sagar Hanumanthu","orcid":"0000-0001-5744-1446","position":4,"is_corresponding":false},{"id":181723,"name":"Shanrun Liu","orcid":null,"position":5,"is_corresponding":false},{"id":659327,"name":"Andrew S. Goldsborough","orcid":null,"position":6,"is_corresponding":false},{"id":392054,"name":"P. Brent Ferrell","orcid":"0000-0003-1140-9154","position":7,"is_corresponding":false},{"id":277438,"name":"Maria B. Grant","orcid":"0000-0002-6470-0255","position":8,"is_corresponding":false},{"id":249182,"name":"Casey T. Weaver","orcid":"0000-0002-2180-1793","position":9,"is_corresponding":false},{"id":339192,"name":"Robert S. Welner","orcid":"0000-0003-2498-9469","position":10,"is_corresponding":false},{"id":339183,"name":"Seth D. Fortmann","orcid":"0000-0002-9554-8751","position":0,"is_corresponding":true}],"reference_count":54,"raw_metadata":null,"created_at":"2026-07-19T02:53:23.001995Z","pmid":"40674215","pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}