Wnt/β-catenin/HNF4α feedback loop facilitates colorectal tumorigenesis and malignancy

Abstract Background: The Wnt/β-catenin signaling pathway is a central regulator of colorectal cancer (CRC) development, yet its downstream targets and mechanistic contributions to tumorigenesis remain poorly defined. Hepatocyte nuclear factor 4 alpha (HNF4α), a transcription factor primarily studied in liver function and hepatocarcinogenesis, has unclear roles in CRC. This study investigates the interplay between HNF4α and Wnt/β-catenin signaling in colorectal carcinogenesis and explores its clinical relevance. Methods: Using bulk RNA sequencing (RNA-seq), single-cell RNA sequencing (scRNA-seq), in vitro and in vivo CRC models, and clinical tumor samples, we assessed HNF4α expression and its regulation by Wnt/β-catenin signaling. Transcriptional activation of HNF4α was evaluated via luciferase reporter assays and chromatin immunoprecipitation. Clinical correlations between HNF4α levels and Wnt/β-catenin activity were analyzed using immunohistochemistry, RNA sequencing, and Spearman’s rank correlation. Statistical significance was determined by Student’s t-test and ANOVA. Results: HNF4α was significantly overexpressed in CRC tissues compared to normal controls and significantly promoted tumor growth in subcutaneous xenograft models using nude mice. Mechanistically, HNF4α was transcriptionally activated by the Wnt/β-catenin/TCF7L1 axis, forming a positive feedback loop that amplified oncogenic Wnt signaling. Clinically, HNF4α expression strongly correlated with Wnt/β-catenin pathway activation in patient samples (r = 0.58, p < 0.0001). Functionally, HNF4α knockdown suppressed CRC cell proliferation and inhibited Wnt-driven tumorigenesis. Conclusions: This study identifies HNF4α as a novel downstream effector of the Wnt/β-catenin pathway and a critical driver of CRC progression. The Wnt/β-catenin/HNF4α feedback loop uncovered here provides mechanistic insights into colorectal carcinogenesis and highlights HNF4α as a potential therapeutic target. These findings may inform strategies to disrupt Wnt signaling hyperactivation in CRC.