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142 Enhanced antitumoral activity of HER2-CAR-Ts in comparison to trastuzumab in a live cell imaging supported 3D assay
  1. Katharina Schaich1,
  2. Gemma Moiset2,
  3. Sophie Vermond2,
  4. Monique Hazenoot2,
  5. Kanstantsin Lashuk1,
  6. Eva Oswald1,
  7. Sanne Holt3 and
  8. Julia Schuler1
  1. 1Charles River Research Services Germany, Freiburg, Germany
  2. 2Charles River, Leiden, Netherlands
  3. 3Merus N.V., Leiden, Netherlands


Background Although most breast cancer patients are treated these days with a curative intention, there are still many patients progressing to metastatic disease. The advent of anti-HER2 therapy has prominently prolonged the time of disease progression and survival for those patients, where a decent proportion is suffering from a HER2+ tumor. Beside classical approaches via antibodies against HER2, the use of Chimeric Antigen Receptor T (CAR-T) cells also in a solid cancer context is getting more and more attention.

Methods In our study we evaluated the efficacy of Trastuzumab and HER2+ targeting CAR-T cells in a panel of human cancer cell lines (SK-OV3, Hs578T and JIMT-1) with different HER2 expression levels. The tumor cells were seeded in the presence or absence of different immune cells (PBMC, monocytes, NK cells or T cells) and cultured as 3D spheroids in a matrix-based system. The tumor growth and if applicable the invasion of the immune cells was measured via fluorescence-based live cell imaging. On the last experiment day, a metabolic read-out (CellTiter-Glo, CTG assay) was performed.

Results Trastuzumab inhibited the tumor growth in a dose-dependent manner in the HER2+ cell line SK-OV3. The efficacy was increased specifically when NK cells were added to the culture. The HER2 positive cell line JIMT-1 was resistant to Trastuzumab treatment, which is in line with published data. The model was derived from a Trastuzumab-refractory patient. Interestingly, the addition of NK cells induced a marked increase of activity in this model as well. The HER2 targeted CART cells eradicated the 3D spheroids of SK-OV3 as well as JIMT-1 in a dose-dependent manner. The untransduced control T cells did not influence the tumor growth at all. The HER2- cell line Hs578T served as a negative control in the Trastuzumab as well as in the CAR-T cell experiments, and proved to be resistant to any treatment in this study. Taken together the 3D live cell imaging platform proved to be a feasible tool for efficacy testing of biologics as well as cellular therapies.

Conclusions Our in house developed HER2 CAR-T cells proved to be specific and effective in eradicating the targeted cancer cells. The mechanism behind the modulated sensitivity of the HER2+ JIMT-1 cells against HER2-targeted treatment will help to shed some light in possible resistance mechanism and hopefully have some translational value for patients suffering from this disease.

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