N6-methyladenosine reader YTHDF3 regulates melanoma metastasis via its 'executor'LOXL3

Area of Science:

• Oncology
• Molecular Biology
• Epigenetics

Background:

• N6-methyladenosine (m6A) modification is implicated in various cancers.
• While m6A writers and erasers influence melanoma, the role of m6A readers like YTHDF proteins remains unclear.

Purpose of the Study:

• To investigate the role of the m6A reader YTH N6-methyladenosine RNA binding protein 3 (YTHDF3) in melanoma.
• To identify and characterize the downstream targets of YTHDF3 in melanoma progression.

Main Methods:

• Quantitative real-time PCR (RT-qPCR), Western blot, and immunohistochemistry were used to assess YTHDF3 and lysyl oxidase-like 3 (LOXL3) expression.
• In vitro and in vivo experiments evaluated the functional impact of YTHDF3 and LOXL3 on melanoma.
• Multi-omics analyses (RNA-seq, MeRIP-seq, RIP-seq) and mass spectrometry identified YTHDF3 targets.
• CRISPR-Cas13b-based epitranscriptome engineering confirmed YTHDF3-LOXL3 interaction.

Main Results:

• YTHDF3 significantly promotes melanoma cell metastasis both in vitro and in vivo.
• LOXL3 was identified as a key downstream target of YTHDF3, with YTHDF3 enhancing LOXL3 protein levels post-transcriptionally.
• Downregulation of LOXL3 inhibited melanoma metastasis, while its overexpression rescued the metastatic phenotype suppressed by YTHDF3 knockdown.

Conclusions:

• The YTHDF3-LOXL3 signaling pathway plays a critical role in melanoma metastasis.
• Targeting the YTHDF3-LOXL3 axis presents a promising therapeutic strategy for inhibiting melanoma progression.