3-Dimensional Patient-Derived Lung Cancer Assays Reveal Resistance to Standards-of-Care Promoted by Stromal Cells but Sensitivity to Histone Deacetylase Inhibitors

Abstract

There is a growing recognition that current preclinical models do not reflect the tumor microenvironment in cellular, biological, and biophysical content and this may have a profound effect on drug efficacy testing, especially in the era of molecular-targeted agents. Here, we describe a method to directly embed low-passage patient tumor-derived tissue into basement membrane extract, ensuring a low proportion of cell death to anoikis and growth complementation by coculture with patient-derived cancer-associated fibroblasts (CAF). A range of solid tumors proved amenable to growth and pharmacologic testing in this 3D assay. A study of 30 early-stage non-small cell lung cancer (NSCLC) specimens revealed high levels of de novo resistance to a large range of standard-of-care agents, while histone deacetylase (HDAC) inhibitors and their combination with antineoplastic drugs displayed high levels of efficacy. Increased resistance was seen in the presence of patient-derived CAFs for many agents, highlighting the utility of the assay for tumor microenvironment-educated drug testing. Standard-of-care agents showed similar responses in the 3D ex vivo and patient-matched in vivo models validating the 3D-Tumor Growth Assay (3D-TGA) as a high-throughput screen for close-to-patient tumors using significantly reduced animal numbers. Mol Cancer Ther; 15(4); 753-63. Ó2016 AACR.

Individual patient oesophageal cancer 3D models for tailored treatment

“…This study and previous work published by our group [5] shows that the role of the stromal component of the TME in influencing the chemo-resistance is both drug- and individual tumour-dependent. This variation is a more accurate reflection of the diverse clinical outcomes to chemotherapy treatment, where OAC patients individually display variable magnitudes of response to individual drugs and the SOC therapy.…”

“…[5] Cellular metabolism and growth was assessed with an alamarBlue ® cell fluorescence assay (ThermoFisher Scientific, Loughborough, UK) and measured on a fluorescent plate reader (excitation 560 nm, emission 588 nm, Flex Station II, Molecular Devices).To enable separate assessment of the co-cultured cells within the 3D-TGA, hMSCs constitutively expressing the red fluorescent protein mCherry were generated (see online supplementary method S5). The mCherry-labelled hMSC component of the 3D-TGA was measured separately by the fluorescent plate reader (excitation 553 nm, emission 613 nm).…”

“…[5] Incorporating extracellular matrix and mesenchymal support restores both direct and paracrine tumour-stroma interactions which are known to influence drug resistance. [6, 7]…”

Antitumor activity of kinetochore-associated protein 2 siRNA against lung cancer patient-derived tumor xenografts

“…Their sensitivities to erlotinib were similar to that of subcutaneous tumors. These results were consistent with previous reports demonstrating the similarities of 3D-culture systems to subcutaneous tumor models using other lung cancer PDXs (41,42).…”

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

“…Patient-derived xenografts were developed using EAC biopsy specimens directly implanted into immuno-deficient mice as previously described 37 . Due to loss of the human stromal compartment in such models, human mesenchymal stem cells (hMSCs) were co-implanted with the xenograft and supplemented before treatment began.…”