Figure 6.

Modeling of patient response to chemotherapy with PDE5i using 3D-TGA. Sensitivity of close-to-patient cells was determined in 3D-TGA, with and without mesenchymal cell co-culture, after 4-day exposure to ECF and vardenafil (PDE5i) drug combinations

(A) Viability curves were generated and IC₅₀ values determined for a cohort of EAC patients’ ECF-treated 3D-TGAs, with (+) and without (−) hMSC support and the addition of PDE5i (n = 15 patient samples from 8 patients). The patient cancer cell clusters were classified as sensitive (green), borderline (orange), or resistant (red) by comparison of IC₅₀ values with the mean peak serum concentrations achieved in patients at the doses used in UK clinical practice. This is marked with an asterisk, where the IC₅₀ drop is significant (CI > 95%, p < 0.05). Tumor regression score (TRG) denotes the chemotherapy response of the patient’s tumor clinically (TRG1-3, sensitive; 4–5, non-responsive).

(B) Overall sensitivity of all the EAC patient samples co-cultured with hMSCs was determined for assays with and without the addition of PDE5i to ECF chemotherapy. Horizontal lines represent mean IC₅₀s.

(C) Gene expression profiling of CAF phenotypes by bulk RNA-seq of 3D-TGA samples from three patients (Oes4R, Oes5R, and Oes7R) with and without hMSC support, and corresponding parent tumors. Heatmap showing expression of 50 well-recognized CAF marker genes. Whereas Oes4R and Oes7R upregulated CAF marker gene expression when hMSC support was added, Oes5R failed to do so (column outlined in blue). See also Figure S2.