Abstract 7013: Methyl-Micro-C: simultaneous high-resolution characterization of three-dimensional chromatin structure and the DNA methylome

Abstract

Abstract Understanding cancer biology allows for the determination of patient prognosis through biomarkers and predicting treatment sensitivity. For many cancer types, genetic changes have been largely characterized, but the contributions of epigenetic changes, which are also implicated in tumorigenesis, are not yet well understood. Three-dimensional chromatin architecture changes occur throughout tumor development, changing the states and interactions between cis-regulatory elements, overall inducing gene dysregulation. Chromatin structure changes have previously been interrogated using Hi-C, a technique which utilizes restriction enzyme-mediated digestion of chromatin. Although this method is useful, it cannot detect chromatin states of regulatory elements with high sensitivity and resolution due to the use of restriction enzymes that give multi-nucleosome-sized fragments. Micro-C instead uses MNase that preferentially digests accessible regulatory elements, resulting in mono, di, and tri-nucleosome fragments, providing higher-resolution chromatin interaction data, indirectly determining which regulatory elements are active and in proximity to one another. However, spatial proximity of genomic regions are only a piece of the chromatin structure epigenetic story. Partially methylated domains, which are gained or altered throughout tumor development, are reported to co-localize with topologically associating domains, suggesting that they can contribute to chromatin structure alterations. Whole methylome sequencing techniques can be used to determine methylation states of these domains and regulatory regions. Recently, enzyme-mediated methylation sequencing has been shown to induce less DNA damage than the canonical whole-genome bisulfite sequencing, providing a more accurate and comprehensive picture of the DNA methylome. Although DNA methylome data alone can provide information about epigenetic status within genomic regions, overlaying chromatin structure changes at these locations can further corroborate findings. In this way, chromatin structure and DNA methylome data together can provide a more complete picture of cancer epigenetics. However, it is very expensive to sequence at the depth necessary to gain useful information. To address this issue, we developed a technique called Methyl-Micro-C to simultaneously interrogate the DNA methylome while observing chromatin structure in the same sample. Methyl-Micro-C integrates the higher resolution Micro-C method and more comprehensive enzyme-mediated methylation sequencing method together to investigate both the chromatin interactions as well as DNA methylation patterns for the same sample simultaneously. Here, we use Methyl-Micro-C with prostate cancer cells to characterize 3D epigenomic mechanisms that drive prostate carcinogenesis. Citation Format: Claire Stevens, Leonardo Gonzalez-Smith, Huan Cao, Suhn K. Rhie. Methyl-Micro-C: simultaneous high-resolution characterization of three-dimensional chromatin structure and the DNA methylome [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 7013.