RUVBL1/2 Blockade Targets YTHDF1 Activity to Suppress m6A-Dependent Oncogenic Translation and Colorectal Tumorigenesis

• Published in: Cancer research (Chicago, Ill.) Vol. 84; no. 17; pp. 2856 - 2872
• Main Authors: Chen, Danyu, Ji, Fenfen, Zhou, Qiming, Cheung, Henley, Pan, Yasi, Lau, Harry C.-H., Liang, Cong, Yang, Zhenjie, Huang, Pingmei, Wei, Qinyao, Cheung, Alvin H.-K., Kang, Wei, Chen, Huarong, Yu, Jun, Wong, Chi Chun
• Format: Journal Article
• Language: English
• Published: United States American Association for Cancer Research 04.09.2024
• Subjects: Adenosine - analogs & derivatives; Adenosine - metabolism; Animals; ATPases Associated with Diverse Cellular Activities - antagonists & inhibitors; ATPases Associated with Diverse Cellular Activities - genetics; ATPases Associated with Diverse Cellular Activities - metabolism; Carcinogenesis - genetics; Carrier Proteins - genetics; Carrier Proteins - metabolism; Cell Line, Tumor; Cell Proliferation; Colorectal Neoplasms - drug therapy; Colorectal Neoplasms - genetics; Colorectal Neoplasms - metabolism; Colorectal Neoplasms - pathology; DNA Helicases - genetics; DNA Helicases - metabolism; Drug Targets; Gastrointestinal Cancers; Gene Expression Regulation, Neoplastic; Gene Regulation; Humans; Mice; Mice, Nude; Oncogenes & Tumor Suppressors; Prognosis; Protein Biosynthesis; RNA-Binding Proteins - genetics; RNA-Binding Proteins - metabolism; Translational Cancer Biology; Xenograft Model Antitumor Assays
• ISSN: 0008-5472; 1538-7445; 1538-7445
• DOI: 10.1158/0008-5472.CAN-23-2081

The N6-methyladenosine (m6A) RNA-binding protein YTHDF1 is frequently overexpressed in colorectal cancer and drives chemotherapeutic resistance. To systematically identify druggable targets in colorectal cancer with high expression of YTHDF1, this study used a CRISPR/Cas9 screening strategy that revealed RUVBL1 and RUVBL2 as putative targets. RUVBL1/2 were overexpressed in primary colorectal cancer samples and represented independent predictors of poor patient prognosis. Functionally, loss of RUVBL1/2 preferentially impaired the growth of YTHDF1-high colorectal cancer cells, patient-derived primary colorectal cancer organoids, and subcutaneous xenografts. Mechanistically, YTHFD1 and RUVBL1/2 formed a positive feedforward circuit to accelerate oncogenic translation. YTHDF1 bound to m6A-modified RUVBL1/2 mRNA to promote translation initiation and protein expression. Coimmunoprecipitation and mass spectrometry identified that RUVBL1/2 reciprocally interacted with YTHDF1 at 40S translation initiation complexes. Consequently, RUVBL1/2 depletion stalled YTHDF1-driven oncogenic translation and nascent protein biosynthesis, leading to proliferative arrest and apoptosis. Ribosome sequencing revealed that RUVBL1/2 loss impaired the activation of MAPK, RAS, and PI3K-AKT signaling induced by YTHDF1. Finally, the blockade of RUVBL1/2 by the pharmacological inhibitor CB6644 or vesicle-like nanoparticle-encapsulated siRNAs preferentially arrested the growth of YTHDF1-expressing colorectal cancer in vitro and in vivo. Our findings show that RUVBL1/2 are potential prognostic markers and druggable targets that regulate protein translation in YTHDF1-high colorectal cancer. Significance: RUVBL1/2 inhibition is a therapeutic strategy to abrogate YTHDF1-driven oncogenic translation and overcome m6A dysregulation in colorectal cancer.