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

Abstract

The M2 pyruvate kinase (PKM2) isoform is upregulated in most cancers and plays a crucial role in regulation of the Warburg effect, which is characterized by the preference for aerobic glycolysis over oxidative phosphorylation for energy metabolism. PKM2 is an alternative-splice isoform of the PKM gene and is a potential therapeutic target. Antisense oligonucleotides (ASO) that switch PKM splicing from the cancer-associated PKM2 to the PKM1 isoform have been shown to induce apoptosis in cultured glioblastoma cells when delivered by lipofection. Here, we explore the potential of ASO-based PKM splice switching as a targeted therapy for liver cancer. A more potent lead constrained-ethyl (cEt)/DNA ASO induced PKM splice switching and inhibited the growth of cultured hepatocellular carcinoma (HCC) cells. This PKM isoform switch increased pyruvate-kinase activity and altered glucose metabolism. In an orthotopic HCC xenograft mouse model, the lead ASO and a second ASO targeting a nonoverlapping site inhibited tumor growth. Finally, in a genetic HCC mouse model, a surrogate mouse-specific ASO induced Pkm splice switching and inhibited tumorigenesis, without observable toxicity. These results lay the groundwork for a potential ASO-based splicing therapy for HCC.

SIGNIFICANCE:

Antisense oligonucleotides are used to induce a change in PKM isoform usage in hepatocellular carcinoma, reversing the Warburg effect and inhibiting tumorigenesis.

• Authors: Wai Kit Ma, Dillon M Voss, Juergen Scharner, Ana S H Costa, Kuan-Ting Lin, Hyun Yong Jeon, John E Wilkinson, Michaela Jackson, Frank Rigo, C Frank Bennett, Adrian R Krainer
• Journal: Cancer research (Cancer Res) Vol. 82 Issue 5 Pg. 900-915 (Mar 01 2022) ISSN: 1538-7445 [Electronic] United States
• PMID: 34921016 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)
• Copyright: ©2021 American Association for Cancer Research.

Chemical References

• Oligonucleotides, Antisense
• Protein Isoforms
• Pyruvate Kinase

Topics

• Alternative Splicing
• Animals
• Carcinogenesis
• Carcinoma, Hepatocellular (genetics, therapy)
• Cell Line, Tumor
• Cell Transformation, Neoplastic (genetics)
• Glycolysis (genetics)
• Humans
• Liver Neoplasms (genetics, therapy)
• Mice
• Oligonucleotides, Antisense (genetics, pharmacology)
• Protein Isoforms (genetics)
• Pyruvate Kinase (genetics, metabolism)