Mesothelioma-associated fibroblasts enhance proliferation and migration of pleural mesothelioma cells via c-Met/PI3K and WNT signaling but do not protect against cisplatin

Our plain-English summary of Mesothelioma-associated fibroblasts enhance proliferation and migration of pleural mesothelioma cells via c-Met/PI3K and WNT signaling but do not protect against cisplatin, by A. Ries, D. Flehberger, A. Slany, et al (Journal of Experimental & Clinical Cancer Research, 2023).

The study

• Authors: A. Ries, D. Flehberger, A. Slany, C. Pirker, J.C. Mader, T. Mohr, K. Schelch, K. Sinn, B. Mosleh, M.A. Hoda, B. Dome, H. Dolznig, G. Krupitza, L. Müllauer, C. Gerner, W. Berger, M. Grusch
• Journal: Journal of Experimental & Clinical Cancer Research
• Published: 2023;42(1):27
• Cited by: 22 articles, as counted by NIH iCite
• Read it: PubMed · Publisher · Free full text

Abstract

Background: Pleural mesothelioma (PM) is an aggressive malignancy with poor prognosis. Unlike many other cancers, PM is mostly characterized by inactivation of tumor suppressor genes. Its highly malignant nature in absence of tumor driving oncogene mutations indicates an extrinsic supply of stimulating signals by cells of the tumor microenvironment (TME). Cancer-associated fibroblasts (CAFs) are an abundant cell type of the TME and have been shown to drive the progression of several malignancies. The aim of the current study was to isolate and characterize patient-derived mesothelioma-associated fibroblasts (Meso-CAFs), and evaluate their impact on PM cells.

Methods: Meso-CAFs were isolated from surgical specimens of PM patients and analyzed by array comparative genomic hybridization, next generation sequencing, transcriptomics and proteomics. Human PM cell lines were retrovirally transduced with GFP. The impact of Meso-CAFs on tumor cell growth, migration, as well as the response to small molecule inhibitors, cisplatin and pemetrexed treatment was investigated in 2D and 3D co-culture models by videomicroscopy and automated image analysis.

Results: Meso-CAFs show a normal diploid genotype without gene copy number aberrations typical for PM cells. They express CAF markers and lack PM marker expression. Their proteome and secretome profiles clearly differ from normal lung fibroblasts with particularly strong differences in actively secreted proteins. The presence of Meso-CAFs in co-culture resulted in significantly increased proliferation and migration of PM cells. A similar effect on PM cell growth and migration was induced by Meso-CAF-conditioned medium. Inhibition of c-Met with crizotinib, PI3K with LY-2940002 or WNT signaling with WNT-C59 significantly impaired the Meso-CAF-mediated growth stimulation of PM cells in co-culture at concentrations not affecting the PM cells alone. Meso-CAFs did not provide protection of PM cells against cisplatin but showed significant protection against the EGFR inhibitor erlotinib.

Conclusions: Our study provides the first characterization of human patient-derived Meso-CAFs and demonstrates a strong impact of Meso-CAFs on PM cell growth and migration, two key characteristics of PM aggressiveness, indicating a major role of Meso-CAFs in driving PM progression. Moreover, we identify signaling pathways required for Meso-CAF-mediated growth stimulation. These data could be relevant for novel therapeutic strategies against PM.

© 2023. The Author(s).

In plain English

This was a laboratory study of cells grown in dishes. It used patients' tumor tissue, and one patient's blood, but did not study asbestos exposure or test treatments or outcomes in patients. Researchers in Vienna grew fibroblast-like cells (a type of connective tissue cell) out of surgical tumor samples from three patients with pleural mesothelioma and called them Meso-CAFs. Genetic tests found none of the chromosome abnormalities seen in mesothelioma cells. Two mesothelioma cell lines, SPC212 and MSTO-211H, grew and moved significantly more when grown with Meso-CAFs than when grown alone. In a 3D collagen gel, the cancer cells showed almost no growth alone but robust growth with Meso-CAFs. Fluid from Meso-CAF cultures also stimulated growth. Of 12 inhibitor drugs, crizotinib, LY294002 and WNT-C59 significantly reduced this growth boost. Meso-CAFs gave no protection against the chemotherapy drug cisplatin. Erlotinib significantly affected cancer cells grown alone but not those grown with Meso-CAFs. The authors concluded that their laboratory study strongly suggests Meso-CAFs have a significant impact on mesothelioma progression. They noted that one fibroblast culture grew too slowly for many experiments.

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) cell-culture study: fibroblasts isolated from patient tumor tissue were characterized genetically and by protein analysis, then grown together with mesothelioma cell lines in 2D and 3D co-cultures
• Who was studied: Fibroblast cultures (Meso109F, Meso125F, VMC59F) grown from surgical tumor specimens of three pleural mesothelioma patients operated on at the Medical University of Vienna. These were combined with the human pleural mesothelioma cell lines SPC212 and MSTO-211H. Normal lung fibroblasts (MRC-5, Wi-38) and a colon cancer fibroblast (CAF-3) were used for comparison. Blood from the patient who donated Meso109F was used for genetic comparison. Dates of sample collection are not given.
• Exposure: No asbestos exposure was studied. The experimental exposures were growth together with Meso-CAFs (1 tumor cell to 100 fibroblasts) or in fluid from Meso-CAF cultures, 12 small-molecule signaling inhibitors, cisplatin (1, 3 and 10 μM in 2D cultures) and pemetrexed (0.3 to 10 μM).
• Main results:
  • Array testing showed no chromosomal abnormalities in the Meso-CAF cultures (Meso109F, Meso125F, VMC59F), unlike the mesothelioma cell lines.
  • In 2D co-culture, Meso109F or Meso125F significantly stimulated growth of both SPC212 and MSTO-211H mesothelioma cells.
  • Co-culture with Meso109F or Meso125F significantly increased the distance migrated by both mesothelioma cell lines.
  • In 3D collagen gels, both mesothelioma cell lines showed almost no growth without Meso-CAFs but robust growth with Meso109F or Meso125F.
  • Conditioned medium (culture fluid) from Meso109F and Meso125F significantly stimulated growth of SPC212 or MSTO-211H cells over 30 hours.
  • Of 12 inhibitors tested, crizotinib (c-Met), LY294002 (PI3K) and WNT-C59 (WNT signaling) significantly reduced the growth stimulation by Meso-CAFs in co-culture. They did not significantly affect mesothelioma cells grown alone.
  • The EGFR inhibitor erlotinib significantly affected mesothelioma cells grown alone but not mesothelioma cells grown with Meso-CAFs.
  • Co-culture with Meso109F or Meso125F did not protect SPC212 or MSTO-211H cells against cisplatin, in either 2D or 3D models.
  • Pemetrexed did not significantly inhibit SPC212 cells at any tested concentration. MSTO-211H cells were significantly inhibited at 1 μM and above, and this effect was slightly reduced in co-culture with Meso-CAFs.
  • Meso109F carried DNA variants of uncertain significance. The FANCL and MSH3 variants were also found in the patient's blood, identifying them as germline (inherited) variants; the EP300 and NTRK3 variants were not found in the blood.
• The authors' conclusion: The authors concluded that their study strongly suggests that Meso-CAFs have a significant impact on pleural mesothelioma progression and sheds light on the signaling pathways responsible for their growth-promoting function.
• Limitations the authors state:
  • Because VMC59F cells proliferated slowly, only a limited number of experiments could be performed with them.
  • Only one of the two mesothelioma cell lines was sensitive to pemetrexed, so the moderate protective effect of Meso-CAFs against pemetrexed must be interpreted with caution.
  • Differences in methods between the published studies used to compare Meso-CAF secreted proteins with those of other cancer fibroblasts could have contributed to the differences found.
  • The candidate marker genes were identified from bulk mRNA data from cultured cells and still need evaluation in patient tissue.

Topics

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

Medical terms in this study, explained in our Lexicon: Mesothelioma, Fibroblast, Pleural Mesothelioma, Cisplatin, Prognosis, Tumor Suppressor Gene, Tumor Microenvironment, Pemetrexed (Alimta).

Related studies

• Primary and hTERT-Transduced Mesothelioma-Associated Fibroblasts but Not Primary or hTERT-Transduced Mesothelial Cells Stimulate Growth of Human Mesothelioma Cells — A. Ries, A. Slany, et al. (2023)
• Fibroblast growth factor receptor inhibition is active against mesothelioma and synergizes with radio- and chemotherapy — K. Schelch, M.A. Hoda, et al. (2014)
• Inhibition of YB-1 phosphorylation enhances cisplatin activity and disrupts cell division in pleural mesothelioma — K. Schelch, N. Maach, et al. (2025)
• 2-Deoxy-glucose Enhances the Effect of Cisplatin and Pemetrexed in Reducing Malignant Pleural Mesothelioma Cell Proliferation But Not Spheroid Growth — I. Gerogianni, E. Pitaraki, et al. (2019)
• Pirfenidone decreases mesothelioma cell proliferation and migration via inhibition of ERK and AKT and regulates mesothelioma tumor microenvironment in vivo — C. Li, V. Rezov, et al. (2018)
• Modulation of Calretinin Expression in Human Mesothelioma Cells Reveals the Implication of the FAK and Wnt Signaling Pathways in Conferring Chemoresistance towards Cisplatin — J. Wörthmüller, V. Salicio, et al. (2019)

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