posted on 2025-08-28, 10:20authored byEunhee Yi, Amit D. Gujar, Dacheng Zhao, Kentaro Suina, Xue Jin, Katharina Pardon, Qinghao Yu, Larisa Kagermazova, Emmanuel E. Korsah, Noah A. Dusseau, Jef D. Boeke, Anton G. Henssen, Roel G.W. Verhaak
<p>Lentiviral transduction causes the depletion of cancer cells with extrachromosomally amplified oncogenes. <b>A,</b> Oncogene amplification status before and after lentiviral transduction. Parental PC3 cells and the transduced PC3 cells that underwent three cycles of lentiviral transduction followed by antibiotics selection were synchronized at metaphase and processed for FISH analysis. MYC probe (red) and chromosomal control probe (green) were used. The number of cells containing <i>MYC</i> amplification was quantified (<i>n</i> = 41). Bar = 10 μm. The lentiviral genome size for each cycle is indicated, along with the name of the antibiotics used in that cycle. Cells were transduced with lentivirus for 48 hours and subjected to antibiotics selection for 1–2 weeks. <b>B,</b> Graphical summary and statistical test of FISH analysis (<i>χ</i><sup>2</sup> test, <i>P</i> < 0.0001). <b>C,</b> Oncogene amplification status before and after lentiviral transduction. The HeLa cell line that acquired methotrexate resistance (HeLa-MTX-Res) underwent two cycles of lentiviral transduction followed by antibiotics selection. Then, the cells were synchronized at metaphase and processed for FISH analysis. A DHFR probe (red) was used. The number of cells containing <i>DHFR</i> amplification was quantified (<i>n</i> = 33). Bar = 10 μm. The lentiviral genome size for each cycle is indicated, along with the names of the antibiotics used in that cycle. Cells were transduced with lentivirus for 48 hours and subjected to antibiotics selection for 1–2 weeks. <b>D,</b> Graphical summary and statistical test of FISH analysis (<i>χ</i><sup>2</sup> test, <i>P</i> = 0.0006).</p>
Funding
Elsa U. Pardee Foundation (EUPF)
National Cancer Institute (NCI)
United States Department of Health and Human Services
Extrachromosomal DNA (ecDNA), a major focal oncogene amplification mode found across cancer, has recently regained attention as an emerging cancer hallmark, with a pervasive presence across cancers. With technical advancements such as high-coverage sequencing and live-cell genome imaging, we can now investigate the behaviors and functions of ecDNA. However, we still lack an understanding of how to eliminate ecDNA. We observed depletion of cells containing ecDNA during lentiviral but not transposon-based transduction, whereas we sought to investigate the mechanism of ecDNA behavior. This discovery may provide critical information on utilizing a lentiviral system in emerging ecDNA research. Additionally, this observation suggests specific sensitivities for cells with ecDNA.
ecDNA is an essential factor in cancer progression. We found that a group of cancer cells with ecDNA is selectively depleted after lentiviral infection. This finding provides promise for ecDNA-specific targeting, suggests the need for caution in using lentivirus, and offers alternative ways to study ecDNA.