Vorinostat differentially alters 3D nuclear structure of cancer and non-cancerous esophageal cells

Abstract

The histone deacetylase (HDAC) inhibitor vorinostat has received significant attention in recent years as an ‘epigenetic’ drug used to treat solid tumors. However, its mechanisms of action are not entirely understood, particularly with regard to its interaction with the aberrations in 3D nuclear structure that accompany neoplastic progression. We investigated the impact of vorinostat on human esophageal epithelial cell lines derived from normal, metaplastic (pre-cancerous), and malignant tissue. Using a combination of novel optical computed tomography (CT)-based quantitative 3D absorption microscopy and conventional confocal fluorescence microscopy, we show that subjecting malignant cells to vorinostat preferentially alters their 3D nuclear architecture relative to non-cancerous cells. Optical CT (cell CT) imaging of fixed single cells showed that drug-treated cancer cells exhibit significant alterations in nuclear morphometry. Confocal microscopy revealed that vorinostat caused changes in the distribution of H3K9ac-marked euchromatin and H3K9me3-marked constitutive heterochromatin. Additionally, 3D immuno-FISH showed that drug-induced expression of the DNA repair gene MGMT was accompanied by spatial relocation toward the center of the nucleus in the nuclei of metaplastic but not in non-neoplastic cells. Our data suggest that vorinostat’s differential modulation of 3D nuclear architecture in normal and abnormal cells could play a functional role in its anti-cancer action.

High-Content Image-Based Single-Cell Phenotypic Analysis for the Testicular Toxicity Prediction Induced by Bisphenol A and Its Analogs Bisphenol S, Bisphenol AF, and Tetrabromobisphenol A in a Three-Dimensional Testicular Cell Co-culture Model

“…Nuclear morphological features have been suggested as useful biomarkers in various adverse cellular events (Eidet et al, 2014;Ikeguchi et al, 1999). Most eukaryotic cellular nuclei are generally round, oval-shaped and smooth, and the changes in nuclear morphology are observed in diverse developmental processes such as the nuclei of spermatocytes (Gan et al, 2013) as well as in pathological conditions such as aging and cancers (Haithcock et al, 2005;Nandakumar et al, 2016;Zink et al, 2004). Recent studies have found that HCA-based quantitative assessment of multiple nuclear parameters was found to be a sensitive marker for detecting early cytotoxic effects (Ramm et al, 2019).…”

Computer Vision and Image Processing: A Paper Review

“…The computer vision frameworks consisted two main groups, e.g., 3D morphological analysis and pixel optimization. The 3D morphological review has been a standard theory for computer image processing and pattern recognition, whereas pixel optimization is related to characterization of pixel morphology, including structural analysis and internal components for a better understanding of vector function [32]. Also, the approach should be performed on relatively large data sets covering many layers of geometrical composition.…”