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SnATAC Sequencing of the aging mouse heart to study chromatin remodeling on single cell level

ABSTRACT: To study chromatin remodelig of the aging mouse heart, hearts from young and gaed mice were taken and nuclei were isolated via FACS. SnATAC Seq was performed using the 10X Genomics kit. Michail Yekelchyk (AG Braun) analysed the data)

ORGANISM(S): Mus musculus

PROVIDER: GSE303973 | GEO | 2026/01/07

REPOSITORIES: GEO

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Project description:Background: A better understanding of the pancreatic adenocarcinoma (PDAC) immune microenvironment is critical to improving outcomes. Myeloid cells are of particular importance for PDAC progression. Circulating monocytes are recruited and hijacked by the tumor to become immune suppressive tumor-associated macrophages. Meanwhile, tissue resident macrophages are innately anti-inflammatory and pro-fibrotic. Open chromatin and transcription factor activity provide source and function data for myeloid cells. Thus, we explore single nuclear assay for transposase accessible chromatin (ATAC) sequencing (snATAC-Seq), a method to analyze open gene promoters and transcription factor binding, as an important means for discerning the myeloid composition in human PDAC tumors. Methods: Frozen pancreatic tissues (benign or PDAC) were digested into single nuclei suspensions. snATAC-Seq and sn-Multiome (ATAC + RNA) libraries were prepared using 10X Chromium technology and sequenced by an Illumina sequencer. Signac was used for preliminary analysis, clustering, and differentially accessible chromatin region identification. snMultiome data was used to validate cell annotations for the snATAC-Seq data. Results: Unique populations comprised the benign and tumor samples. Myeloid cell transcription factor activities were higher in tumor samples than benign pancreatic samples, enabling us to further stratify myeloid populations in PDAC tumors. GO enrichment analysis showed myeloid cells can activate immune response in tumors. Subcluster analysis revealed eight distinct myeloid populations. GO enrichment demonstrated unique functions of different myeloid populations, including IL-1b signaling (recruited monocytes) and intracellular protein transport (dendritic cells). Conclusions: These data suggest snATAC-Seq as a method for retrospective analysis of frozen human pancreatic tissues to identify myeloid populations. Understanding the source specific potential function of myeloid cells is important for patient prognosis in PDAC.

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SnATAC Sequencing of the aging mouse heart to study chromatin remodeling on single cell level

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