The Chick Embryo Xenograft Model for Malignant Pleural Mesothelioma: A Cost and Time Efficient 3Rs Model for Drug Target Evaluation

Our plain-English summary of The Chick Embryo Xenograft Model for Malignant Pleural Mesothelioma: A Cost and Time Efficient 3Rs Model for Drug Target Evaluation, by S.E. Barnett, A. Herrmann, L. Shaw, et al (Cancers, 2022).

The study

• Authors: S.E. Barnett, A. Herrmann, L. Shaw, E.N. Gash, H. Poptani, J.J. Sacco, J.M. Coulson
• Journal: Cancers
• Published: 2022;14(23)
• Cited by: 11 articles, as counted by NIH iCite
• Read it: PubMed · Publisher · Free full text

Abstract

Malignant pleural mesothelioma (MPM) has limited treatment options and poor prognosis. Frequent inactivation of the tumour suppressors BAP1, NF2 and P16 may differentially sensitise tumours to treatments. We have established chick chorioallantoic membrane (CAM) xenograft models of low-passage MPM cell lines and protocols for evaluating drug responses. Ten cell lines, representing the spectrum of histological subtypes and tumour suppressor status, were dual labelled for fluorescence/bioluminescence imaging and implanted on the CAM at E7. Bioluminescence was used to assess viability of primary tumours, which were excised at E14 for immunohistological staining or real-time PCR. All MPM cell lines engrafted efficiently forming vascularised nodules, however their size, morphology and interaction with chick cells varied. MPM phenotypes including local invasion, fibroblast recruitment, tumour angiogenesis and vascular remodelling were evident. Bioluminescence imaging could be used to reliably estimate tumour burden pre- and post-treatment, correlating with tumour weight and Ki-67 staining. In conclusion, MPM-CAM models recapitulate important features of the disease and are suitable to assess drug targets using a broad range of MPM cell lines that allow histological or genetic stratification. They are amenable to multi-modal imaging, potentially offering a time and cost-efficient, 3Rs-compliant alternative to rodent xenograft models to prioritise candidate compounds from in vitro studies.

In plain English

This was a laboratory study developing a chick embryo chorioallantoic membrane (CAM) xenograft model for malignant pleural mesothelioma (MPM), a cancer the authors describe in their introduction as mainly caused by environmental exposure to asbestos. Ten human MPM cell lines, covering epithelioid, biphasic and sarcomatoid subtypes and combinations of BAP1, NF2 and P16 inactivation, were labelled with luciferase and ZsGreen and implanted on the CAM at embryonic day 7, with tumour nodules examined at day 14. All ten lines formed vascularised nodules; engraftment among surviving embryos ranged from 43% to 85%, and was 75% to 85% for epithelioid lines (mean 80%). Nodule size, shape, invasion, chick fibroblast infiltration and vascularisation differed between lines. Bioluminescence correlated with dissected tumour weight (Spearman r = 0.39, p = 0.0015, n = 63) and with Ki-67 score in three biphasic lines (Pearson r = 0.96, p = 0.0023). In one cell line only, MSTO-211H, MMP9 and VEGF transcripts rose significantly in CAM nodules (n = 6) compared with that line's 2D cultures (n = 3). The authors concluded the model offers a 3Rs-compliant complement to rodent xenografts.

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: experimental laboratory study developing and characterising a chick embryo chorioallantoic membrane (CAM) xenograft model using human mesothelioma cell lines
• Who was studied: Ten human malignant pleural mesothelioma cell lines (MSTO-211H from ATCC; MESO-7T, -8T, -12T, -23T, -29T and MPM#2, #24, #26, #34 from MesobanK UK), implanted on fertilised hen's eggs at the University of Liverpool
• Exposure: implantation of dual-labelled MPM cells (0.5, 1 or 2 million cells per egg; 2 million in the main experiments) onto the traumatised CAM at embryonic day 7; no anticancer drug was tested, only vehicle (PBS) yolk-sac injections
• Main results:
  • Among embryos surviving to E14, engraftment ranged from 43% to 85% across the ten cell lines, and from 75% to 85% for the epithelioid lines (mean 80%)
  • Mean dissected nodule weight was 5-7 mg for many cell lines, but 15-17 mg for MESO-8T, MESO-12T and MSTO-211H
  • Across individual xenografts from all cell lines, bioluminescence signal correlated with dissected tumour weight
  • In nodules from three biphasic cell lines (MPM#2, MPM#26, MSTO-211H), Ki-67 scores correlated closely with bioluminescence
  • In the MSTO-211H cell line only, MMP9 and VEGF transcript levels were increased in 3D CAM nodules (n = 6) compared with 2D in vitro cultures (n = 3)
  • Total vessel length around tumour-bearing CAM was lower than on control non-tumour-bearing CAM
  • In a vehicle-only (PBS) timeline experiment with MSTO-211H, 80% of embryos survived E10 to E14 and bioluminescence rose a mean of 4.40-fold (SD 1.82) in 7 tumours
  • Preliminary cisplatin experiments showed no demonstrable activity in the mesothelioma CAM xenografts
  • Histology and immunofluorescence showed alpha-SMA-positive chick fibroblast-like cells within the nodules, distinct from the ZsGreen-labelled MPM cells
• The authors' conclusion: The authors concluded that MPM-CAM xenografts can be efficiently established across histological subtypes and tumour suppressor backgrounds, mimic stromal and vascular interactions missing from cell culture, and can be monitored by bioluminescence imaging, potentially offering a time- and cost-efficient, 3Rs-compliant component of the drug development pathway
• Limitations the authors state:
  • The authors state the model cannot fully replace rodent or other higher organism models and is complementary
  • The authors state that, as with non-humanised rodent flank xenografts, CAM xenografts of cancer cell lines cannot be used to test immune modulators or inhibitors at E14
  • The authors note xenografts derived from cell lines established in 2D culture recapitulate original tumour histology less well than fresh patient-derived samples
  • The authors note repeated handling and luciferin injection for longitudinal imaging may reduce embryo survival
  • The authors note the usefulness of bioluminescence imaging could be limited by transduction efficiency

Topics

Medical terms in this study, explained in our Lexicon: Pleural Mesothelioma, Prognosis, BAP1, Histological Subtype, Fibroblast, Angiogenesis, Ki-67.

Related studies

• Methodology for generating chorioallantoic membrane patient-derived xenograft (CAM-PDX) models of pleural mesothelioma and performing preclinical imaging for the translation of cancer studies and drug screening — J. Schulze, S. Barnett, et al. (2025)
• Photodynamic drug delivery enhancement in tumours does not depend on leukocyte-endothelial interaction in a human mesothelioma xenograft model — Y. Wang, J.Y. Perentes, et al. (2012)
• Nitric Acid Leaching for Magnesium Extraction from Asbestos Ore Waste: From DoE to Predictive Modeling and Cost-Efficient Optimization — N.S. Ivanov, O.S. Kholkin, et al. (2025)
• Influence of clinical benefit and cost-effectiveness on reimbursement decisions for lung cancer drugs in Spain — P. Nieto-Gómez, C. Castaño-Amores, et al. (2026)
• Preclinical Efficacy and Proteomic Prediction of Molecular Targets for s-cal14.1b and s-cal14.2b Conotoxins with Antitumor Capacity in Xenografts of Malignant Pleural Mesothelioma — A. Luna-Nophal, F. Díaz-Castillo, et al. (2025)
• Evaluation of the cost-effectiveness of cytoreductive surgery and hyperthermic intraperitoneal chemotherapy (peritonectomy) at the St George Hospital peritoneal surface malignancy program — T.C. Chua, S. Martin, et al. (2010)

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