Organoid model of mammographic density displays a higher frequency of aberrant colony formations with radiation exposure

Abstract

Motivation: Aberrant three-dimensional (3D) colony organization of premalignant human mammary epithelial cells (HMECs) is one of the indices of dysplasia. An experiment has been designed where the stiffness of the microenvironment, in 3D culture, has been set at either low or high level of mammographic density (MD) and the organoid models are exposed to 50 cGy X-ray radiation. This study utilizes published bioinformatics tools to quantify the frequency of aberrant colony formations by the combined stressors of stiffness and X-ray exposure. One of the goals is to develop a quantitative assay for evaluating the risk factors associated with women with high MD exposed to X-ray radiation.

Results: Analysis of 3D colony formations indicate that high stiffness, within the range of high MD, and X-ray radiation have an approximately additive effect on increasing the frequency of aberrant colony formations. Since both stiffness and X-ray radiation are DNA-damaging stressors, the additive effect of these stressors is also independently validated by profiling activin A-secreted protein. Secretion of activin A is known to be higher in tissues with a high MD as well as tumor cells. In addition, we show that increased stiffness of the microenvironment also induces phosphorylation of γH2AX-positive foci. The study uses two HMECs derived from a diseased tissue (e.g. MCF10A) and reduction mammoplasty of normal breast tissue (e.g. 184A1) to further demonstrate similar traits in the frequency of aberrant colony organization.

Supplementary information: Supplementary data are available at Bioinformatics online.