Quantitative high throughput screening using a primary human three-dimensional organotypic culture predicts in vivo efficacy.

Methods Development
Paradigms and Technologies

Abstract

The tumour microenvironment contributes to cancer metastasis and drug resistance. However, most high throughput screening (HTS) assays for drug discovery use cancer cells grown in monolayers. Here we show that a multilayered culture containing primary human fibroblasts, mesothelial cells and extracellular matrix can be adapted into a reliable 384- and 1,536-multi-well HTS assay that reproduces the human ovarian cancer (OvCa) metastatic microenvironment. We validate the identified inhibitors in secondary in vitro and in vivo biological assays using three OvCa cell lines: HeyA8, SKOV3ip1 and Tyk-nu. The active compounds directly inhibit at least two of the three OvCa functions: adhesion, invasion and growth. In vivo, these compounds prevent OvCa adhesion, invasion and metastasis, and improve survival in mouse models. Collectively, these data indicate that a complex three-dimensional culture of the tumour microenvironment can be adapted for quantitative HTS and may improve the disease relevance of assays used for drug screening.

Authors

Kenny, Hilary A; Lal-Nag, Madhu; White, Erin A; Shen, Min; Chiang, Chun-Yi; Mitra, Anirban K; Zhang, Yilin; Curtis, Marion; Schryver, Elizabeth M; Bettis, Sam; Jadhav, Ajit; Boxer, Matthew B; Li, Zhuyin; Ferrer-Alegre, Marc; Lengyel, Ernst;

Keywords

  • Animals
  • Antineoplastic Agents/ chemistry
  • Antineoplastic Agents/ pharmacology
  • Benzophenanthridines/ chemistry
  • Benzophenanthridines/ pharmacology
  • Biguanides/ chemistry
  • Biguanides/ pharmacology
  • Cantharidin/ chemistry
  • Cantharidin/ pharmacology
  • Cell Adhesion/ drug effects
  • Cell Movement/ drug effects
  • Cell Proliferation/ drug effects
  • Coculture Techniques
  • Epithelial Cells/ drug effects
  • Epithelial Cells/ metabolism
  • Epithelial Cells/ pathology
  • Escin/ chemistry
  • Escin/ pharmacology
  • Extracellular Matrix/ drug effects
  • Extracellular Matrix/ metabolism
  • Female
  • Fibroblasts/ drug effects
  • Fibroblasts/ metabolism
  • Fibroblasts/ pathology
  • High-Throughput Screening Assays/ instrumentation
  • High-Throughput Screening Assays/ methods
  • Humans
  • Inhibitory Concentration 50
  • Isoquinolines/ chemistry
  • Isoquinolines/ pharmacology
  • Mice
  • Mice, Nude
  • Ovarian Neoplasms/ drug therapy
  • Ovarian Neoplasms/ metabolism
  • Ovarian Neoplasms/ pathology
  • Primary Cell Culture
  • Prochlorperazine/ chemistry
  • Prochlorperazine/ pharmacology
  • Tomatine/ chemistry
  • Tomatine/ pharmacology
  • Tumor Microenvironment/ drug effects
  • Xenograft Model Antitumor Assays

External Links