Gene Expression Profiling and Epigenetic Alterations in RAW 264.7 Cells Following Treatment with Lipopolysaccharide and Methylcobalamin (MCB) [ATAC-seq]

Methylcobalamin (MCB), an active form of Vitamin B12, demonstrates significant anti-inflammatory and neuroprotective properties by modulating pro-inflammatory cytokines such as TNF-α, IL-1β, and IL-6 (Chopra et al., 2013; Scalabrino et al., 2019). This modulation could potentially ameliorate the hyperinflammatory response known as the "cytokine storm" observed in COVID-19 patients, thereby reducing severity and improving clinical outcomes (Tanaka et al., 2020; Scalabrino, 2005). Beyond its immunomodulatory effects, methylcobalamin supports neuronal health, a critical factor given the neurological complications associated with COVID-19 (Scalabrino et al., 2017; Zhu et al., 2021). Although further clinical trials are necessary to confirm its efficacy, methylcobalamin presents a promising adjunctive therapy for managing the inflammatory and neurological symptoms of COVID-19. However, the precise molecular mechanisms underlying MCB's anti-inflammatory effects remain to be fully elucidated. In this study, we aim to investigate the transcriptional and epigenetic mechanisms through which MCB functions as an anti-inflammatory agent. To explore the transcriptomic and epigenetic mechanisms through which methylcobalamin (MCB) modulates the inflammatory response induced by lipopolysaccharide (LPS) in macrophages, we conducted RNA sequencing (RNA-seq), assay for transposase-accessible chromatin using sequencing (ATAC-seq), and Cleavage Under Targets and Tagmentation (CUT & Tag) on the RAW 264.7 cell line. Initially, we conducted a comprehensive gene expression profiling analysis using ATAC-seq data from WT RAW 264.7 cells treated with DMSO, LPS, LPS-MCB, and MCB for 12 hours. Comparative ATAC-seq analysis of RNA-seq data for DMSO, LPS, LPS-MCB and MCB group.