Individual patient oesophageal cancer 3D models for tailored treatment

Abstract

BackgroundA model to predict chemotherapy response would provide a marked clinical benefit, enabling tailored treatment of oesophageal cancer, where less than half of patients respond to the routinely administered chemotherapy.MethodsCancer cells were established from tumour biopsies taken from individual patients about to undergo neoadjuvant chemotherapy. A 3D-tumour growth assay (3D-TGA) was developed, in which cancer cells were grown with or without supporting mesenchymal cells, then subjected to chemo-sensitivity testing using the standard chemotherapy administered in clinic, and a novel emerging HDAC inhibitor, Panobinostat.RESULTSIndividual patients cancer cells could be expanded and screened within a clinically applicable timescale of 3 weeks. Incorporating mesenchymal support within the 3D-TGA significantly enhanced both the growth and drug resistance profiles of the patients cancer cells. The ex vivo drug response in the presence, but not absence, of mesenchymal cells accurately reflected clinical chemo-sensitivity, as measured by tumour regression grade. Combination with Panobinostat enhanced response and proved efficacious in otherwise chemo-resistant tumours.ConclusionsThis novel method of establishing individual patient oesophageal cancers in the laboratory, from small endoscopic biopsies, enables clinically-relevant chemo-sensitivity testing, and reduces use of animals by providing more refined in vitro models for pre-screening of drugs. The 3D-TGA accurately predicted chemo-sensitivity in patients, and could be developed to guide tailored patient treatment. The incorporation of mesenchymal cells as the stromal cell component of the tumour micro-environment had a significant effect upon enhancing chemotherapy drug resistance in oesophageal cancer, and could prove a useful target for future drug development.

Modulation of the tumour promoting functions of cancer associated fibroblasts by phosphodiesterase type 5 inhibition increases the efficacy of chemotherapy in human preclinical models of esophageal adenocarcinoma

“…The standard-of-care for locally advanced esophageal adenocarcinoma treatment includes neoadjuvant chemotherapy before resection. Cancer is confirmed by pre-treatment biopsy and this tissue can be used to grow the patient’s cancer epithelial cells ex vivo 30 . We have used cancer cells harvested at this time point, as well as matched cells established from post-neoadjuvant chemotherapy surgical specimens, to investigate the potential for PDE5i treatment to enhance the efficacy of standard-of-care chemotherapy (epirubicin, cisplatin and 5-FU; ECF).…”

“…We have used cancer cells harvested at this time point, as well as matched cells established from post-neoadjuvant chemotherapy surgical specimens, to investigate the potential for PDE5i treatment to enhance the efficacy of standard-of-care chemotherapy (epirubicin, cisplatin and 5-FU; ECF). Ex vivo cancer cells (confirmed by FACS) were grown with human mesenchymal stem cells (hMSC) in a 3D matrix to form a 3D-Tumor Growth Assay (3D-TGA) as previously described 30 (Figure 6A,B) . We have previously shown the importance of the stromal compartment in 3D-TGA as the ex vivo drug response in the presence, but not absence, of mesenchymal cells accurately reflected clinical chemo-sensitivity, as measured by tumor regression grade (TRG) 30 .…”

“…siRNA-mediated silencing of PDE5 was performed using INTERFERin transfection reagent and two commercially available siRNA sequences (detailed in supplementary methods). Endoscopic tumour biopsies (REC: 10/H0401/80) and fresh surgical specimens (REC: 08/H0403/37) were collected with informed consent from patients at Nottingham University Hospitals NHS Trust in 2014-15, and used in accordance with National Research Ethics Service approval to generate oesophageal cancer cell-lines (detailed in supplementary methods) as previously described 30 .…”

“…The individual patient’s epithelial cells were established, characterised, and then co-cultured in the 3D-TGA +/-human mesenchymal stem cells (hMSC) 30 . Following exposure to drug combinations at human tissue-relevant concentrations, the cell viability was assessed using the alamarBlue® assay (ThermoFisher Scientific, Loughborough, UK).…”

“…Viability curves were generated and IC 50 values calculated using GraphPad Prism 5 software (San Diego, CA, USA). These were compared with the mean peak serum concentration achieved in patients for each chemotherapeutic agent to evaluate chemotherapeutic response as previously reported 30 . For full details see supplementary methods.…”