ASO-Based PKM Splice-Switching Therapy Inhibits Hepatocellular Carcinoma Growth

Area of Science:

• Biochemistry
• Molecular Biology
• Oncology

Background:

• The M2 pyruvate kinase (PKM2) isoform is upregulated in most cancers, driving the Warburg effect for energy metabolism.
• PKM2 is an alternative-splice isoform of the PKM gene and represents a potential therapeutic target for cancer.
• Antisense oligonucleotides (ASO) can switch PKM splicing from PKM2 to PKM1, inducing apoptosis in cancer cells.

Purpose of the Study:

• To explore the potential of ASO-based PKM splice switching as a targeted therapy for liver cancer (hepatocellular carcinoma, HCC).

Main Methods:

• Developed and tested constrained-ethyl (cEt)/DNA ASOs to induce PKM splice switching in HCC cells and mouse models.
• Assessed the impact of PKM isoform switching on pyruvate-kinase activity and glucose metabolism.
• Evaluated ASO efficacy in inhibiting tumor growth in orthotopic HCC xenograft and genetic HCC mouse models.

Main Results:

• A lead cEt/DNA ASO successfully induced PKM splice switching, inhibited growth of cultured HCC cells, and increased pyruvate-kinase activity.
• The lead ASO and a second ASO targeting a different site inhibited tumor growth in an orthotopic HCC xenograft mouse model.
• A mouse-specific ASO induced Pkm splice switching and inhibited tumorigenesis in a genetic HCC mouse model with no observable toxicity.

Conclusions:

• ASO-based PKM splice switching is a promising targeted therapy for HCC.
• This approach reverses the Warburg effect and inhibits tumor growth.
• Preclinical studies demonstrate the potential of ASO-based splicing therapy for HCC with a favorable safety profile.