Validation of chemoresistance phenotypes in pleural mesothelioma across 2D, 3D, and in vivo models

Our plain-English summary of Validation of chemoresistance phenotypes in pleural mesothelioma across 2D, 3D, and in vivo models, by H. Shi, S.P. Selvamani, R. Zelei, et al (Scientific Reports, 2026).

The study

• Authors: H. Shi, S.P. Selvamani, R. Zelei, L. Zhuang, D. Wang, B. Johnson, V. Dharwal, D. Xu, Y. Wang, T. Rutland, S. Klebe, S. Kao, A. Linton, E. Hosseini-Beheshti, Y.Y. Cheng
• Journal: Scientific Reports
• Published: 2026;16(1)
• Cited by: 2 articles, as counted by NIH iCite
• Read it: PubMed · Publisher · Free full text

Abstract

Pleural mesothelioma (PM) is a highly aggressive cancer with limited treatment efficacy and poor prognosis. Conventional two-dimensional (2D) culture models fail to replicate the tumour microenvironment (TME), limiting their translational relevance. Here, we establish a three-dimensional (3D) spheroid model to investigate chemotherapy resistance across different PM subtypes. Compared to 2D cultures, 3D spheroids display enhanced resistance to cisplatin-pemetrexed, with elevated IC₅₀ values, reduced apoptosis, and altered cell cycle profiles. Seahorse metabolic analysis of 3D spheroids demonstrate a suppressed metabolic phenotype, characterised by reduced oxidative phosphorylation (OCR). However, the glycolytic capacity was not upregulated, consistent with the hypoxic and nutrient-limited conditions observed in mesothelioma lesions. In parallel, molecular profiling identifies subtype-specific miRNA signatures that closely align with patient-derived datasets. Proteomic analysis of 3D cultures identifies upregulation of PI3K/AKT and Notch/VEGF signalling, implicating these pathways in treatment resistance. Histological assessment of xenografts further confirms 3D model fidelity in capturing tumour fibrosis, necrosis, and response to therapy. These findings position the 3D spheroid system as a robust and physiologically relevant platform for modelling drug resistance and guiding therapeutic development in PM.

© 2026. The Author(s).

In plain English

This laboratory study of pleural mesothelioma cells did not examine asbestos exposure. The authors grew mesothelioma cells of the epithelioid, biphasic and sarcomatoid subtypes as flat layers (2D) or as three-dimensional clusters called spheroids (3D), then treated them with cisplatin-pemetrexed chemotherapy. Spheroids needed higher doses: the dose that halved viability was 1732 µM for sarcomatoid, 215.9 µM for epithelial and 145.2 µM for biphasic spheroids, compared with 9.87 to 14.64 µM in 2D cultures. Treatment raised early apoptosis in 2D cultures, but spheroids showed only a 5% increase, and a downward trend in their oxygen use was not statistically significant. In mice with abdominal MSTO tumours, spheroid-derived tumours did not shrink after chemotherapy. Responses were 12.5% in the 3D group versus 28.5% in the 2D group. Survival did not discernibly differ, but follow-up ended when all mice were euthanised at day 32. Spheroid-derived tumours showed more fibrosis. The authors concluded that 3D spheroids better mimic tumour behaviour seen in patients. They noted that the abdominal model differs from the pleural cavity and that no patient tumour samples were used.

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 study comparing 2D and 3D spheroid cultures of mesothelioma cells, plus a mouse xenograft experiment
• Who was studied: A panel of pleural mesothelioma cells from the three histological subtypes. Commercial lines were H2052, H2452, H28, H226, MSTO, MM05 and VMC23. Patient-derived cells established at the Asbestos and Dust Diseases Research Institute were 1157, 1137, 1180, 1506, 2174 and 2359. Cells 1157 (epithelioid), MSTO (biphasic) and 1137 (sarcomatoid) were used for the detailed tests. Female SCID mice were implanted intraperitoneally with MSTO cells.
• Exposure: No asbestos exposure was studied. The exposure was chemotherapy: cisplatin-pemetrexed (and, in cell culture, cisplatin, pemetrexed, vinorelbine and gemcitabine alone). Mice received four cycles of cisplatin 5 mg/kg and pemetrexed 200 mg/kg over 28 days.
• Main results:
  • The cisplatin-pemetrexed dose halving viability (IC50) in 3D spheroids was 1732 µM for sarcomatoid, 215.9 µM for epithelial and 145.2 µM for biphasic cells.
  • In 2D cultures, sensitivity was similar across subtypes. Table 1 gives IC50 values of 9.87 µM (epithelial), 12.87 µM (biphasic) and 14.64 µM (sarcomatoid).
  • After treatment, early apoptosis in 2D cultures rose from 11.8% to 31.5% (1157), 10.7% to 29.3% (MSTO) and 8.7% to 21.6% (1137). 3D spheroids showed only a 5% increase.
  • In non-epithelioid 3D spheroids, treatment significantly increased miR-21 (fold-change about 2.45) and miR-30e (fold-change about 1.94).
  • Treatment significantly lowered oxygen consumption rate in 2D cells.
  • In 3D spheroids, respiration showed a downward trend after treatment that was not statistically significant.
  • In mice, chemotherapy reduced the size of 2D-derived tumours but not 3D-derived tumours.
  • Average tumour weight was 0.34 g for 3D-derived tumours and 0.18 g for 2D-derived tumours.
  • The response to chemotherapy was 12.5% in mice implanted with 3D spheroids and 28.5% in mice implanted with 2D cells.
  • Survival showed no discernible difference between the two groups of mice. All mice were euthanised at day 32, and the authors stated that this absence of a difference reflects the study design rather than equivalent long-term outcomes.
  • On pathologist assessment, 57% of 3D-derived tumours showed a poor or no response to chemotherapy, while 71% of 2D-derived tumours showed an intermediate response.
  • Fibrosis was mild in 2D-derived tumours and moderate in 3D-derived tumours.
  • 3D-derived tumours, particularly after treatment, showed significant upregulation of proteins linked to the PI3K/AKT pathway and to the VEGF/Notch pathway.
• The authors' conclusion: The authors concluded that 3D spheroid models better mimic tumour behaviour seen in patients and reflect clinical resistance patterns more accurately than 2D cultures.
• Limitations the authors state:
  • The mouse tumours were grown in the abdomen (intraperitoneal). Drug distribution and stromal makeup there differ from the pleural cavity and may influence treatment responses.
  • All mice were euthanised at day 32, so survival was not followed beyond that point.
  • The findings were not directly validated in patient tumour samples, because tissue availability was limited.
  • Extracellular lactate levels were not directly measured.

Topics

The National Library of Medicine indexes this study under subjects our site covers: Mesothelioma.

Medical terms in this study, explained in our Lexicon: Pleural Mesothelioma, Prognosis, Chemotherapy, Cisplatin, Pemetrexed (Alimta), Apoptosis, Mesothelioma, Molecular Profiling.

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