Poster abstracts
Poster number 17 submitted by Chayan Bhattacharya
Epigenetic And LncRNA Regulation In Vascular Cells Under Cancer Therapy-Induced Stress
Chayan Bhattacharya (Center for Molecular Medicine and Genetics Wayne State University, Detroit, MI), Miguel Nieto-Hernandez (Center for Molecular Medicine and Genetics,Wayne State University, Detroit, MI), Sudarshan Anand (Knight Cancer Institute, Oregon Health and Science University, Portland, OR.), Cristina Espinosa-Diez (Center for Molecular Medicine and Genetics,Wayne State University, Detroit, MI)
Abstract:
Genotoxic stress from cancer treatments disrupts vascular homeostasis, and can lead to lasting changes in epigenetic modifications, such as DNA methylation and long-non-coding RNAs (lncRNAs), which influence vascular cell fate. We hypothesize that cancer therapy leads to chronic vascular dysfunction due to this epigenetic reprogramming, which may affect the expression of critical vascular health regulators like lncRNAs. To assess the differential DNA methylation under genotoxic stress, we subjected human vascular endothelial cells (HUVECs) to ionizing radiation, followed by reduced representation bisulfite sequencing (RRBS). Our data revealed differentially hypomethylated and hypermethylated gene promoters, including 5% from lncRNA genes. To assess the role of these lncRNAs in vascular function, we employed an in-silico pipeline to identify novel lncRNA candidates, considering factors such as the number of isoforms, their coding potential, conservation across mammals, neighboring genomic location, and basal expression in blood vessels and vascularized tissues. From an initial pool of 52 lncRNAs potentially altered by DNA methylation in response to ionizing radiation, we identified 5 lncRNA candidates that met our selection criteria. To validate the potential changes in expression driven by DNA methylation alterations, we performed RT-qPCR for these lncRNA candidates in endothelial cells treated with doxorubicin, a genotoxic stressor. We identified three lncRNA candidates— DIO3OS, LINC00354, and RP5-998H6—in response to genotoxic stress that changed expression in response to doxorubicin, suggesting that they can be integral components of a broader regulatory network that helps endothelial cells adapt to and survive DNA damage. Our findings suggest that cancer therapies induce significant epigenetic reprogramming in vascular endothelial cells, particularly affecting lncRNAs. Future experiments will further validate the functional roles of these lncRNA candidates through gain and loss of function studies.
Keywords: LncRNAs, Vascular Biology, genotoxic stress