{"doi":"10.1101/2025.05.15.653154","title":"Single-Cell RNA Sequencing and Inferred Protein Activity Analysis Reveal a Distinct Tumor Phenotype in Early-Onset Colorectal Cancer Patients","abstract":"Colorectal cancer (CRC) diagnosed before age 50 years (early-onset CRC, EO-CRC) is rising at an alarming rate, yet its molecular and microenvironmental drivers remain poorly understood. EO-CRC is highly heterogeneous, and while subtle differences from late-onset CRC (LO-CRC) have been reported, their full extent remains unresolved due to the limited scope of previous studies. Here, we integrate public data with in-house clinical samples profiled by single-cell RNA sequencing (scRNA-seq) and multiplex immunofluorescence (mIF) to compare EO-CRC and LO-CRC. Additionally, we employ gene regulatory network-based protein activity inference (VIPER), enabling a more precise characterization of key regulatory proteins driving tumor-stroma interactions. Our analysis reveals that EO-CRC and LO-CRC have a largely similar immune composition, challenging previous reports of an \"immune-cold\" phenotype in EO-CRC. However, we identify distinct stromal differences, including a significant enrichment of fibroblasts in EO-CRC. Notably, we define a previously unrecognized epithelial subpopulation in EO-CRC, marked by high expression of toll-like receptor 4 (TLR4) and C-C chemokine receptor type 5 (CCR5)-key mediators of inflammation-driven tumor progression and fibroblast recruitment. These findings suggest that EO-CRC may be driven by a tumor-intrinsic inflammatory phenotype with enhanced stromagenesis, providing new insights into potential mechanisms underlying its increasing incidence in young adults.","journal":"bioRxiv (Cold Spring Harbor Laboratory)","year":2025,"id":566091,"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":0,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9394,"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":266898,"name":"Aleksandar Obradović","orcid":"0000-0002-8009-0186","position":1,"is_corresponding":false},{"id":1469930,"name":"Luka Jovanović","orcid":"0000-0001-9402-7391","position":2,"is_corresponding":false},{"id":59320,"name":"Yosuke Mitani","orcid":"0000-0003-1603-0004","position":3,"is_corresponding":false},{"id":904149,"name":"Tianxia Li","orcid":"0000-0001-8735-7309","position":4,"is_corresponding":false},{"id":860515,"name":"Soobeom Lee","orcid":"0000-0001-7352-0654","position":5,"is_corresponding":false},{"id":1368999,"name":"Dechokyab De","orcid":null,"position":6,"is_corresponding":false},{"id":1470233,"name":"Casidhe-Nicole Bethancourt","orcid":null,"position":7,"is_corresponding":false},{"id":1013867,"name":"Alice E. Shin","orcid":"0000-0002-6474-8312","position":8,"is_corresponding":false},{"id":991031,"name":"Karen J. Dunbar","orcid":"0000-0003-0505-000X","position":9,"is_corresponding":false},{"id":261866,"name":"Francesco Cambuli","orcid":"0000-0002-8237-7121","position":10,"is_corresponding":false},{"id":307547,"name":"Christopher J. Lengner","orcid":"0000-0002-0574-5189","position":11,"is_corresponding":false},{"id":434621,"name":"Ning Li","orcid":"0000-0001-5997-5556","position":12,"is_corresponding":false},{"id":1470234,"name":"Caitlin Rogers","orcid":null,"position":13,"is_corresponding":false},{"id":1284149,"name":"Jennifer Ho","orcid":"0000-0002-3703-8251","position":14,"is_corresponding":false},{"id":1470235,"name":"Viktoria Jakubikova","orcid":null,"position":15,"is_corresponding":false},{"id":1470236,"name":"Emma Blystone","orcid":null,"position":16,"is_corresponding":false},{"id":581605,"name":"Yoanna S Pumpalova","orcid":"0000-0001-5520-9264","position":17,"is_corresponding":false},{"id":1127615,"name":"Beatrice Dionigi","orcid":null,"position":18,"is_corresponding":false},{"id":483,"name":"Sanja Vicković","orcid":"0000-0003-0985-9885","position":19,"is_corresponding":false},{"id":62844,"name":"Chia‐Wei Cheng","orcid":null,"position":20,"is_corresponding":false},{"id":59332,"name":"Joel Gabre","orcid":"0000-0002-5347-3341","position":21,"is_corresponding":false},{"id":1469929,"name":"David Cohen","orcid":"0000-0002-4108-6846","position":0,"is_corresponding":true}],"reference_count":39,"raw_metadata":null,"created_at":"2026-07-19T02:56:36.440192Z","pmid":"40462964","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":[]}