A dog oviduct-on-a-chip model of serous tubal intraepithelial carcinoma

Marcia De Almeida Monteiro Melo Ferraz,Séverine Le Gac,Bastien Venzac,Nucharin Songsasen,Jennifer Beth Nagashima

doi:10.1038/s41598-020-58507-4

Abstract

Ovarian cancer is the fifth cause of cancer-related mortality in women, with an expected 5-year survival rate of only 47%. High-grade serous carcinoma (HGSC), an epithelial cancer phenotype, is the most common malignant ovarian cancer. It is known that the precursors of HGSC originate from secretory epithelial cells within the Fallopian tube, which first develops as serous tubal intraepithelial carcinoma (STIC). Here, we used gene editing by CRISPR-Cas9 to knock out the oncogene p53 in dog oviductal epithelia cultured in a dynamic microfluidic chip to create an in vitro model that recapitulated human STIC. Similar to human STIC, the gene-edited oviduct-on-a-chip, exhibited loss of cell polarization and had reduced ciliation, increased cell atypia and proliferation, with multilayered epithelium, increased Ki67, PAX8 and Myc and decreased PTEN and RB1 mRNA expression. This study provides a biomimetic in vitro model to study STIC progression and to identify potential biomarkers for early detection of HGSC.

Highlights

Ovarian cancer is the fifth cause of cancer-related mortality in women, with an expected 5-year survival rate of only 47%
Past research on High-grade serous carcinoma (HGSC) has used human tumor cells, but it has been hampered by the inefficiency of platting and sub-culturing of Fallopian tube epithelium, a process that involves a long period of fibroblast contamination reduction[7]
Here we introduce a novel fabrication methodology relying on partial curing of PDMS parts from 3D printed molds, followed by bonding to the intermediate thermoplastic membrane, using only an oven and a desiccator

Summary

Introduction

Ovarian cancer is the fifth cause of cancer-related mortality in women, with an expected 5-year survival rate of only 47%. We used gene editing by CRISPR-Cas[9] to knock out the oncogene p53 in dog oviductal epithelia cultured in a dynamic microfluidic chip to create an in vitro model that recapitulated human STIC. Past research on HGSC has used human tumor cells, but it has been hampered by the inefficiency of platting and sub-culturing of Fallopian tube epithelium, a process that involves a long period of fibroblast contamination reduction[7] These processes result in the selection of specific cell populations, which lack tumor molecular characteristics, mutations and intra-patient heterogeneity[7,8,9]. Organ-on-a-chip platforms are microscale advanced in vitro models that have brought novel capabilities to engineer levels of cell organization, differentiation and interaction that cannot be readily achieved by conventional static 2D cultures[32]. Gene editing of oviductal cells is limited in platforms like the EVATAR, which employ explants instead of cells layers[38]

Methods

Results

Conclusion