July 1, 2019
The drugs were added 1 hour after plating the organoids using the Tecan D300e Digital Dispenser (Tecan). Nutlin-3 (Cayman Chemical, catalog no. 10004372), Niraparib (Selleckchem, catalog no. S2741), AZD4547 (ApeXbio, catalog no. A8250), everolimus (LC Laboratories, catalog no. E4040), vemurafenib (Selleckchem, catalog no. S1267), and alpelisib (LC Laboratories, catalog no. A4477) were dissolved in DMSO. Cisplatin (Sigma, catalog no. C2210000), carboplatin (Sigma, catalog no. C2538), and cetuximab (obtained from hospital pharmacy) were dissolved in PBS containing 0.3% Tween-20, which was required to dispense these drugs using the HP printer. All wells were normalized for solvent used. DMSO percentage never exceeded 1%, PBS/Tween-20 percentage never exceeded 2%. Drug exposure was performed in triplicate for each concentration shown. For a layout of the drug screen, see Supplementary Fig. S12
Previous studies have described that tumor organoids can capture the diversity of defined human carcinoma types. Here, we describe conditions for long-term culture of human mucosal organoids. Using this protocol, a panel of 31 head and neck squamous cell carcinoma (HNSCC)-derived organoid lines was established. This panel recapitulates genetic and molecular characteristics previously described for HNSCC. Organoids retain their tumorigenic potential upon xenotransplantation. We observe differential responses to a panel of drugs including cisplatin, carboplatin, cetuximab, and radiotherapy in vitro. Additionally, drug screens reveal selective sensitivity to targeted drugs that are not normally used in the treatment of patients with HNSCC. These observations may inspire a personalized approach to the management of HNSCC and expand the repertoire of HNSCC drugs. SIGNIFICANCE: This work describes the culture of organoids derived from HNSCC and corresponding normal epithelium. These tumoroids recapitulate the disease genetically, histologically, and functionally. In vitro drug screening of tumoroids reveals responses to therapies both currently used in the treatment of HNSCC and those not (yet) used in clinical practice.See related commentary by Hill and D'Andrea, p. 828.This article is highlighted in the In This Issue feature, p. 813. Š2019 American Association for Cancer Research.
Driehuis, E; Kolders, S; Spelier, S; LĂľhmussaar, K; Willems, SM; Devriese, LA; de Bree, R; de Ruiter, EJ; Korving, J; Begthel, H; van Es, JH; Geurts, V; He, GW; van Jaarsveld, RH; Oka, R; Muraro, MJ; ViviĂŠ, J; Zandvliet, MMJM; Hendrickx, APA; Iakobachvili, N; Sridevi, P; Kranenburg, O; van Boxtel, R; Kops, GJPL; Tuveson, DA; Peters, PJ; van Oudenaarden, A; Clevers, H;
Journal: Cancer Discov Pages: 852-871