Multi-lineage reprogramming and NOTCH-associated signaling remodeling characterize the aggressive biology of early-onset colorectal cancer
- 1Department of Gastrointestinal Surgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology
- 2Department of Health Toxicology, Key Laboratory for Environment and Health, School of Public Health, Tongji Medical College, Huazhong University of Science and Technology
- 3Department of Chronic and Noncommunicable Disease Prevention and Control, Jiaxing Center for Disease Control and Prevention
Abstract
Introduction: Early-onset colorectal cancer (EOCRC), defined as colorectal cancer diagnosed before 50 years of age, represents an emerging global health challenge with rapidly increasing incidence. Although EOCRC exhibits distinct clinical and molecular characteristics, the cellular ecosystem and mechanisms underlying its aggressive behavior remain incompletely understood.
Methods: Here, we integrated seven independent single-cell RNA sequencing cohorts comprising 473,939 high-quality cells from 174 colorectal cancer (CRC) patients (39 EOCRC and 135 average-onset CRC [AOCRC]) to construct a comprehensive cellular landscape across age groups. After Harmony-based integration, nine major cellular lineages were identified, and malignant epithelial cells were characterized using chromosome-level copy number variation inference. We systematically compared malignant and stromal compartments, followed by malignant-state characterization, trajectory analysis, and cell -cell communication modeling using Slingshot, CellChat, and NicheNet.
Results: EOCRC displayed extensive remodeling of the tumor ecosystem rather than isolated epithelial alterations. Malignant epithelial cells in EOCRC showed increased enrichment of inflammation-associated and Cycling (S/DNA damage response [DDR]) states. In parallel, proliferating endothelial cells and MKI67-positive proliferative cancer-associated fibroblasts were expanded in EOCRC, indicating stromal-state remodeling. Copy number variation analyses demonstrated that genomic instability differences between EOCRC and AOCRC were primarily explained by differential microsatellite instability (MSI) status distribution rather than age-related genomic alterations. Cell -cell communication analyses revealed selective remodeling of epithelial-stromal signaling networks, with recurrent involvement of NOTCH-associated mediators. NicheNet prioritized NOTCH-associated ligands (DLL4, JAG1) among candidate stromal-derived regulators of EOCRC transcriptional programs. A refined five-gene NOTCH-epithelial-mesenchymal transition (EMT) panel (HEY1, ZEB1, TWIST1, VIM, FN1) was associated with EOCRC-specific poor survival across two independent cohorts, and in silico perturbation further suggested that increased HEY1 activity may reinforce the proliferative state enriched in EOCRC.
Discussion: Together, our study establishes EOCRC as a multicellular ecosystem characterized by coordinated malignant, stromal, and signaling reprogramming. These findings identify NOTCH signaling as a potential age-specific prognostic association that warrants functional validation in EOCRC.
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BibTeX
@article{shi2026multilineage,
title = {{Multi-lineage reprogramming and NOTCH-associated signaling remodeling characterize the aggressive biology of early-onset colorectal cancer}},
author = {Hongda Shi and Li Qiao and Qianqian Su and Hui Gao and Yang Cao},
journal = {Frontiers in Oncology},
year = {2026},
month = oct,
doi = {10.3389/fonc.2026.1962237},
url = {https://www.frontiersin.org/articles/10.3389/fonc.2026.1962237/full},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - Multi-lineage reprogramming and NOTCH-associated signaling remodeling characterize the aggressive biology of early-onset colorectal cancer
AU - Hongda Shi
AU - Li Qiao
AU - Qianqian Su
AU - Hui Gao
AU - Yang Cao
PY - 2026
DA - 2026/10/22
JO - Frontiers in Oncology
DO - 10.3389/fonc.2026.1962237
UR - https://www.frontiersin.org/articles/10.3389/fonc.2026.1962237/full
AB - Introduction: Early-onset colorectal cancer (EOCRC), defined as colorectal cancer diagnosed before 50 years of age, represents an emerging global health challenge with rapidly increasing incidence. Although EOCRC exhibits distinct clinical and molecular characteristics, the cellular ecosystem and mechanisms underlying its aggressive behavior remain incompletely understood. Methods: Here, we integrated seven independent single-cell RNA sequencing cohorts comprising 473,939 high-quality cells from 174 colorectal cancer (CRC) patients (39 EOCRC and 135 average-onset CRC [AOCRC]) to construct a comprehensive cellular landscape across age groups. After Harmony-based integration, nine major cellular lineages were identified, and malignant epithelial cells were characterized using chromosome-level copy number variation inference. We systematically compared malignant and stromal compartments, followed by malignant-state characterization, trajectory analysis, and cell -cell communication modeling using Slingshot, CellChat, and NicheNet. Results: EOCRC displayed extensive remodeling of the tumor ecosystem rather than isolated epithelial alterations. Malignant epithelial cells in EOCRC showed increased enrichment of inflammation-associated and Cycling (S/DNA damage response [DDR]) states. In parallel, proliferating endothelial cells and MKI67-positive proliferative cancer-associated fibroblasts were expanded in EOCRC, indicating stromal-state remodeling. Copy number variation analyses demonstrated that genomic instability differences between EOCRC and AOCRC were primarily explained by differential microsatellite instability (MSI) status distribution rather than age-related genomic alterations. Cell -cell communication analyses revealed selective remodeling of epithelial-stromal signaling networks, with recurrent involvement of NOTCH-associated mediators. NicheNet prioritized NOTCH-associated ligands (DLL4, JAG1) among candidate stromal-derived regulators of EOCRC transcriptional programs. A refined five-gene NOTCH-epithelial-mesenchymal transition (EMT) panel (HEY1, ZEB1, TWIST1, VIM, FN1) was associated with EOCRC-specific poor survival across two independent cohorts, and in silico perturbation further suggested that increased HEY1 activity may reinforce the proliferative state enriched in EOCRC. Discussion: Together, our study establishes EOCRC as a multicellular ecosystem characterized by coordinated malignant, stromal, and signaling reprogramming. These findings identify NOTCH signaling as a potential age-specific prognostic association that warrants functional validation in EOCRC.
SN - 2234-943X
ER -