Reprogramming of the m⁶A Epitranscriptome Drives Triptolide-Induced Reproductive Toxicity in HTR-8/SVneo Cells

Area of Science:

• Reproductive Biology
• Molecular Pharmacology
• Epigenetics

Background:

• Triptolide (TPL), derived from Tripterygium wilfordii Hook F (TwHF), exhibits diverse bioactivities but is limited by reproductive toxicity.
• The precise molecular mechanisms underlying TPL-induced reproductive toxicity are not well understood.

Purpose of the Study:

• To elucidate the molecular mechanism of TPL-induced trophoblast cell injury.
• To investigate the role of N⁶-methyladenosine (m⁶A) modification in TPL's reproductive toxicity.

Main Methods:

• Integrated analysis of MeRIP-seq and mRNA-seq data.
• Experimental validation using HTR-8/SVneo trophoblast cells.
• Molecular docking to assess TPL binding to m⁶A regulators.

Main Results:

• TPL significantly altered m⁶A modification and gene expression, with 1774 genes showing hypomethylation and mRNA upregulation.
• Affected genes were enriched in reproduction-related pathways like autophagy, DNA damage response, and apoptosis.
• TPL directly bound to m⁶A regulators, causing specific demethylation of genes such as E2F1 and PPP1CC.

Conclusions:

• TPL induces trophoblast cell injury and reproductive toxicity through a novel post-transcriptional mechanism involving disruption of m⁶A modification.
• Perturbation of m⁶A pathways affects essential trophoblast functions, contributing to observed toxicity.