Decoding a cancer-relevant splicing decision in the RON proto-oncogene using high-throughput mutagenesis

Area of Science:

• Molecular Biology
• Genetics
• Cancer Research

Background:

• Aberrant RNA splicing is a known driver of human diseases, particularly cancer.
• Understanding the regulatory mechanisms of alternative splicing is crucial for disease intervention.

Purpose of the Study:

• To investigate the cis-regulatory landscape controlling alternative splicing of MST1R (RON) exon 11.
• To identify mutations impacting RON exon 11 splicing and their correlation with cancer.
• To elucidate the role of heterogeneous nuclear ribonucleoprotein H (HNRNPH) in RON splicing.

Main Methods:

• High-throughput screening of randomly mutated minigenes.
• Mathematical modeling of splicing kinetics.
• Integrated analysis of iCLIP and synergy data.
• Correlation analysis with patient mutation data.

Main Results:

• Over 1000 mutations affecting RON exon 11 skipping were identified, leading to the pathological RON∆165 isoform.
• Identified mutations showed a correlation with alternative splicing patterns in cancer patients.
• Heterogeneous nuclear ribonucleoprotein H (HNRNPH) was identified as a key regulator of RON splicing in both healthy and cancerous tissues.
• Specific HNRNPH binding sites were pinpointed, and cooperative binding was shown to regulate RON exon 11 splicing.

Conclusions:

• The study decodes the cis-regulatory landscape of RON exon 11 splicing and its pathological implications in cancer.
• HNRNPH plays a critical role in regulating RON splicing, offering potential therapeutic targets.
• Findings provide insights into mutation-driven splicing dysregulation in cancer.