Optimization of tumor spheroid model in mesothelioma and lung cancers and anti-cancer drug testing in H2052/484 spheroids

Our plain-English summary of Optimization of tumor spheroid model in mesothelioma and lung cancers and anti-cancer drug testing in H2052/484 spheroids, by D.A.J. Gendre, E. Ameti, W. Karenovics, et al (Oncotarget, 2021).

The study

• Authors: D.A.J. Gendre, E. Ameti, W. Karenovics, N. Perriraz-Mayer, F. Triponez, V. Serre-Beinier
• Journal: Oncotarget
• Published: 2021;12(24):2375-2387
• Cited by: 23 articles, as counted by NIH iCite
• Read it: PubMed · Publisher · Free full text

Abstract

Advanced lung cancers and mesothelioma remain incurable diseases. Despite some promising new therapy strategies, predicting whether an individual patient will be sensitive to a given therapy is challenging. The purpose of this study is to establish and evaluate the efficiency of a three-dimensional spheroid model of human thoracic cancer in predicting the efficacy of drugs. Human mesothelioma and lung tumor spheroids were established from cell lines and primary cells derived from the patient. The growth kinetics and cell viability of microtumors were assessed using spheroid size and intracellular ATP level. The sensitivity of the mesothelioma spheroids to the cisplatin or cisplatin/pemetrexed combination was determined. We determined that studying the kinetics of the spheroid growth for 15 days after seeding 1000 cells/well in a 96-well plate was optimal. Monitoring the growth kinetic and intracellular ATP of spheroids allowed the identification of early changes in spheroid viability. Finally, we validated this model by measuring a dose-dependent reduction in the cell viability of mesothelioma H2052/484 spheroids treated with both first-line treatments, cisplatin and the cisplatin/pemetrexed combination. In conclusion, we have developed a three-dimensional spheroid model of thoracic tumor cells useful for tailoring the medical treatment to the specific characteristics of each patient.

Copyright: © 2021 Gendre et al.

In plain English

This was a laboratory study at the University Hospitals of Geneva. It grew small three-dimensional clumps of cancer cells, called spheroids, to see whether they could be used to measure how tumor cells respond to drugs. It did not study asbestos exposure. The authors used three malignant pleural mesothelioma cell lines (H2052, H2052/484 and H2452) and lung adenocarcinoma cells grown from surgical samples from two patients (LuCa1, LuCa61, LuCa62). The cell types differed in how tightly they formed spheroids. Seeding 1000 cells per well was enough to form a spheroid 100 μm across by day 3. Except for LuCa61, spheroids grew without falling apart from day 3 to day 15. Levels of ATP, a measure of living cells, rose with cell number and spheroid size. Drugs were tested only on H2052/484 mesothelioma spheroids. A 3-hour treatment with cisplatin or a cisplatin/pemetrexed combination reduced their growth and ATP levels in a dose-dependent way. The calculated IC50 was 1.21 µM for cisplatin and 3.18 µM/12.72 µM for the combination. The authors concluded that the model can be used to characterize anti-cancer drug effects on patient-derived cells.

We wrote this plain-English summary from the full article with the help of AI, and a second AI model checked each sentence against the article. It is not the authors' wording; the abstract and the article are their own account.

What the study did and found

• Design: Laboratory (in vitro) study developing and testing a three-dimensional tumor spheroid cell-culture model, including drug testing on spheroids
• Who was studied: Cultured cells, not people. Three human malignant pleural mesothelioma cell lines: H2052 and H2452, purchased from American Type Culture Collection, and H2052/484, obtained as described previously. Lung adenocarcinoma cells established in the authors' laboratory from surgically removed tumors of patients at the University Hospitals of Geneva: LuCa1 from a 79-year-old woman, and LuCa61 and LuCa62 from a 72-year-old man. The study was approved by the Swiss ethics committee in 2018.
• Exposure: No asbestos exposure was studied. The exposures were the chemotherapy drugs cisplatin (50, 100 or 200 μM) and a cisplatin/pemetrexed combination (50/200, 100/400 or 200/800 μM, a 1:4 ratio). These were applied for 3 hours to H2052/484 mesothelioma spheroids 3 days after seeding. Spheroids were then followed for 17 days, or for 24 hours in the IC50 experiments using cisplatin from 0 to 100 μM.
• Main results:
  • Mesothelioma H2052 and H2052/484 cells formed tight, regular spheroids 200 to 300 μm across after 13 days. H2452 mesothelioma cells and LuCa61 lung adenocarcinoma cells did not compact and stayed loosely aggregated.
  • Seeding 1000 cells per well was enough to form a spheroid with a diameter of 100 μm by day 3.
  • For all cell populations except LuCa61, spheroids increased in size without disaggregation from day 3 to day 15.
  • Adding a sonication step to the ATP test gave ATP readings about 2-fold higher for the lung adenocarcinoma cells. The ratios were 2.74 for LuCa1 (p = 0.0040), 1.81 for LuCa61 and 2.62 for LuCa62 (p = 0.0056). Readings were only slightly higher for the mesothelioma spheroids.
  • After 3 days, ATP levels in H2052/484 and LuCa1 spheroids were linearly related to the number of viable cells and to spheroid diameter. The signal saturated with spheroids formed from 25000 cells or more.
  • In H2052/484 mesothelioma spheroids, growth and ATP levels fell in a dose-dependent way with both cisplatin and the cisplatin/pemetrexed combination compared with untreated spheroids. The reduction started at 50 μM cisplatin and at 50 μM/200 μM of the combination.
  • Spheroids treated with 200 µM cisplatin/800 µM pemetrexed showed no increase in size or ATP level. The authors said this suggests these concentrations completely blocked growth and viability.
  • The authors stated that the combination seemed to affect growth and viability of H2052/484 spheroids more efficiently than cisplatin alone.
  • In H2052/484 spheroids, with ATP measured 24 hours after a 3-hour treatment, the calculated IC50 was 1.21 µM for cisplatin and 3.18 µM/12.72 µM for the cisplatin/pemetrexed combination.
• The authors' conclusion: The authors concluded that they had established a simple, reliable in vitro 3D tumor spheroid model that can be used to characterize the efficacy of anti-cancer drugs on primary cells from patients with mesothelioma or lung cancer. In this study, drug testing was done only on the H2052/484 mesothelioma cell line.

Topics

Medical terms in this study, explained in our Lexicon: Mesothelioma, Lung Cancer, Cisplatin, Pemetrexed (Alimta).

Related studies

• Narciclasine as a Novel Treatment for Lung Cancer and Malignant Pleural Mesothelioma: Insights from 3D Tumor Spheroid Models — S. Jiang, S. Berkcan, et al. (2025)
• Sound-based assembly of a microcapillary network in a saturn-like tumor model for drug testing — N. Di Marzio, P. Ananthanarayanan, et al. (2022)
• 3-Dimensional mesothelioma spheroids provide closer to natural pathophysiological tumor microenvironment for drug response studies — H. Shi, E.M. Rath, et al. (2022)
• Changes in global gene expression associated with 3D structure of tumors: an ex vivo matrix-free mesothelioma spheroid model — H. Kim, Y. Phung, et al. (2012)
• First-in-Human, Multicenter, Phase I Dose-Escalation and Expansion Study of Anti-Mesothelin Antibody-Drug Conjugate Anetumab Ravtansine in Advanced or Metastatic Solid Tumors — R. Hassan, G.R. Blumenschein, et al. (2020)
• Paclitaxel-loaded expansile nanoparticles enhance chemotherapeutic drug delivery in mesothelioma 3-dimensional multicellular spheroids — H. Lei, S.C. Hofferberth, et al. (2015)

Browse more research in Oncotarget, or return to medical research.