Primary and hTERT-Transduced Mesothelioma-Associated Fibroblasts but Not Primary or hTERT-Transduced Mesothelial Cells Stimulate Growth of Human Mesothelioma Cells

Our plain-English summary of Primary and hTERT-Transduced Mesothelioma-Associated Fibroblasts but Not Primary or hTERT-Transduced Mesothelial Cells Stimulate Growth of Human Mesothelioma Cells, by A. Ries, A. Slany, C. Pirker, et al (Cells, 2023).

The study

• Authors: A. Ries, A. Slany, C. Pirker, J.C. Mader, D. Mejri, T. Mohr, K. Schelch, D. Flehberger, N. Maach, M. Hashim, M.A. Hoda, B. Dome, G. Krupitza, W. Berger, C. Gerner, K. Holzmann, M. Grusch
• Journal: Cells
• Published: 2023;12(15)
• Cited by: 4 articles, as counted by NIH iCite
• Read it: PubMed · Publisher · Free full text

Abstract

Pleural mesothelioma (PM) is an aggressive malignancy that develops in a unique tumor microenvironment (TME). However, cell models for studying the TME in PM are still limited. Here, we have generated and characterized novel human telomerase reverse transcriptase (hTERT)-transduced mesothelial cell and mesothelioma-associated fibroblast (Meso-CAF) models and investigated their impact on PM cell growth. Pleural mesothelial cells and Meso-CAFs were isolated from tissue of pneumothorax and PM patients, respectively. Stable expression of hTERT was induced by retroviral transduction. Primary and hTERT-transduced cells were compared with respect to doubling times, hTERT expression and activity levels, telomere lengths, proteomes, and the impact of conditioned media (CM) on PM cell growth. All transduced derivatives exhibited elevated hTERT expression and activity, and increased mean telomere lengths. Cell morphology remained unchanged, and the proteomes were similar to the corresponding primary cells. Of note, the CM of primary and hTERT-transduced Meso-CAFs stimulated PM cell growth to the same extent, while CM derived from mesothelial cells had no stimulating effect, irrespective of hTERT expression. In conclusion, all new hTERT-transduced cell models closely resemble their primary counterparts and, hence, represent valuable tools to investigate cellular interactions within the TME of PM.

In plain English

This laboratory study generated and characterized new human telomerase reverse transcriptase (hTERT)-transduced cell models for research on the tumour microenvironment of pleural mesothelioma. Primary pleural mesothelial cells (NP2) were isolated from a pneumothorax surgery specimen and mesothelioma-associated fibroblasts (Meso-CAFs; Meso109F, Meso125F) from pleural mesothelioma tumour tissue, all collected at the Medical University of Vienna. Stable hTERT expression was introduced by retroviral transduction, and the transduced derivatives were compared with their primary parent cells for morphology, doubling time, hTERT expression and telomerase activity, telomere length and proteomes. All transduced cells showed increased hTERT expression and telomerase activity and longer telomeres, with unchanged morphology; mean doubling times fell, significantly for NP2 (162.1 h to 43.4 h) and for Meso125F (180.5 h to 109.1 h). Proteomes of transduced and primary cells correlated closely (for example R = 0.83 for NP2-hT+ versus NP2 supernatant). Conditioned medium from primary and hTERT-transduced Meso-CAFs stimulated growth of the mesothelioma cell lines SPC212 and MSTO-211H, whereas conditioned medium from NP2, NP2-hT+ or Met5A produced no statistically significant growth stimulation, although the authors noted a slight trend for Met5A. Asbestos was mentioned only as background.

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: in vitro cell culture study generating and characterizing hTERT-transduced human cell models, with conditioned-medium growth assays
• Who was studied: Primary human pleural mesothelial cells (NP2) from a pneumothorax surgery specimen and mesothelioma-associated fibroblasts (Meso109F, Meso125F) from pleural mesothelioma patients, surgery at the Medical University of Vienna; comparison cell lines Met5A, MRC-5, Wi38, and mesothelioma lines MSTO-211H and SPC212
• Exposure: Retroviral transduction with hTERT-EGFP, and treatment of mesothelioma cells with conditioned media from the mesothelial cell and Meso-CAF cultures; no asbestos exposure was studied
• Main results:
  • Conditioned medium from both primary and hTERT-transduced Meso-CAFs stimulated growth of the mesothelioma cell lines SPC212 and MSTO-211H to the same extent, while conditioned medium from Met5A, NP2 or NP2-hT+ showed no significant growth stimulation
  • The authors noted a slight trend, but no significant growth increase, with Met5A conditioned medium
  • Doubling times were reduced in all three hTERT-transduced models, with statistical significance for NP2-hT+ and Meso125F-hT+ compared with their primary parents
  • Mean doubling times in hours were Meso109F 161.5, Meso109F-hT+ 114.4, Meso125F 180.5, Meso125F-hT+ 109.1, NP2 162.1, NP2-hT+ 43.4, Met5A 40.9, MRC-5 227.9, Wi38 181.3
  • Proteomes of hTERT-transduced mesothelial cells closely matched their primary parent: Pearson R 0.83 (supernatant), 0.87 (cytoplasmic) and 0.79 (nuclear) for NP2-hT+ versus NP2
  • Correlations were lower between NP2 and Met5A: 0.66 (supernatant), 0.71 (cytoplasmic) and 0.56 (nuclear)
  • Meso-CAF derivatives also matched their primary counterparts (R 0.83, 0.92, 0.89 for Meso109F-hT+ vs Meso109F; 0.88, 0.91, 0.87 for Meso125F-hT+ vs Meso125F)
  • All transduced cells showed strongly increased hTERT mRNA and telomerase activity and increased mean telomere lengths; Met5A had the highest telomerase activity but very short telomeres
  • Growth factors and proteases including CXCL12, IGF2, WNT5B and MMP2 were either found only in Meso-CAF conditioned medium or at significantly lower levels in mesothelial cell conditioned medium
• The authors' conclusion: The authors concluded that the new hTERT-transduced cell models are valuable tools for investigating cellular interactions involved in the development, progression and drug response of pleural mesothelioma
• Limitations the authors state:
  • The authors stated that the lower HGF levels in mesothelial cell secretomes were probably not sufficient to explain the difference in growth stimulation, and that several factors combined, rather than one molecule, likely mediate the effect
  • The authors noted that gene expression differences of Meso-CAFs compared with mesothelioma cells, normal lung fibroblasts and CAFs from other tumours were not explored in this study

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, Mesothelial Cells, Pleural Mesothelioma, Tumor Microenvironment, Pneumothorax.

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