Comprehensive analysis of full-length transcripts reveals novel splicing abnormalities and oncogenic transcripts in liver cancer

PLoS Genet. 2022 Aug 4;18(8):e1010342. doi: 10.1371/journal.pgen.1010342. eCollection 2022 Aug.

Abstract

Genes generate transcripts of various functions by alternative splicing. However, in most transcriptome studies, short-reads sequencing technologies (next-generation sequencers) have been used, leaving full-length transcripts unobserved directly. Although long-reads sequencing technologies would enable the sequencing of full-length transcripts, the data analysis is difficult. In this study, we developed an analysis pipeline named SPLICE and analyzed cDNA sequences from 42 pairs of hepatocellular carcinoma (HCC) and matched non-cancerous livers with an Oxford Nanopore sequencer. Our analysis detected 46,663 transcripts from the protein-coding genes in the HCCs and the matched non-cancerous livers, of which 5,366 (11.5%) were novel. A comparison of expression levels identified 9,933 differentially expressed transcripts (DETs) in 4,744 genes. Interestingly, 746 genes with DETs, including the LINE1-MET transcript, were not found by a gene-level analysis. We also found that fusion transcripts of transposable elements and hepatitis B virus (HBV) were overexpressed in HCCs. In vitro experiments on DETs showed that LINE1-MET and HBV-human transposable elements promoted cell growth. Furthermore, fusion gene detection showed novel recurrent fusion events that were not detected in the short-reads. These results suggest the efficiency of full-length transcriptome studies and the importance of splicing variants in carcinogenesis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alternative Splicing / genetics
  • Carcinogenesis / genetics
  • Carcinoma, Hepatocellular* / genetics
  • DNA Transposable Elements
  • Hepatitis B virus / genetics
  • High-Throughput Nucleotide Sequencing / methods
  • Humans
  • Liver Neoplasms* / genetics
  • RNA Splicing / genetics
  • Transcriptome / genetics

Substances

  • DNA Transposable Elements

Grants and funding

This work was supported by grant 18H02680 from the Japan Society for the Promotion of Science (JSPS) to AF, and by grant JP21km0908001 from Japan Agency for Medical Research and Development (AMED) to AF, and by grant from Takeda Science Foundation to AF. The funders had no role in study design, data collection, and analysis, decision to publish, or preparation of the manuscript.