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Single-molecule DNA methylation reveals unique epigenetic identity profiles of T helper cells | LC 25


Biography

Chloe Goldsmith is an epigenetics researcher investigating the role of 5-methylcytosine (5mC) and 5-hydroxymethylcytosine (5hmC) in immune cell identity and function. Her work focuses on the influence of DNA modifications on immune responses in autoimmune diseases and cancer immunity. By exploring the epigenetic regulation of immune cells, Chloe aims to uncover mechanisms driving immune dysfunction and discover potential therapeutic targets for disease.

Abstract

Both identity and plasticity of CD4 T helper (Th) cells are regulated in part by epigenetic mechanisms. However, a method that reliably and readily profiles DNA base modifications is still needed to finely study Th cell differentiation.

Cytosine methylation in CpG context (5mCpG) and cytosine hydroxymethylation (5hmCpG) are DNA modifications that identify stable cell phenotypes, but their potential to characterize intermediate cell transitions has not yet been evaluated.

To assess transition states in Th cells, we developed a method to profile Th cell identity using Cas9-targeted single-molecule nanopore sequencing. Targeting as few as 10 selected genomic loci, we were able to distinguish major in vitro polarized murine T cell subtypes, as well as intermediate phenotypes, by their native DNA 5mCpG patterns. Moreover, by using off-target sequences, we were able to infer transcription factor activities relevant to each cell subtype.

Detection of 5mCpG and 5hmCpG was validated on intestinal Th17 cells escaping transforming growth factor β control, using single-molecule adaptive sampling. A total of 21 differentially methylated regions mapping to the 10-gene panel were identified in pathogenic Th17 cells relative to their non-pathogenic counterpart.

Hence, our data highlight the potential to exploit native DNA methylation profiling to study physiological and pathological transition states of Th cells.

Authors: Chloe Goldsmith

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