Lens Epithelial Explants Treated with Vitreous Humor Undergo Alterations in Chromatin Landscape with Concurrent Activation of Genes Associated with Fiber Cell Differentiation and Innate Immune Response
Padula, Stephanie L.
Tangeman, Jared A.
Wagner, Brad D.
O’Connell, Michael J.
Jaquish, Tycho J.
Palko, Raye K.
Mantz, Courtney J.
Lovicu, Frank J.
Lens epithelial explants are comprised of lens epithelial cells cultured in vitro on their native basement membrane, the lens capsule. Biologists have used lens epithelial explants to study many different cellular processes including lens fiber cell differentiation. In these studies, fiber differentiation is typically measured by cellular elongation and the expression of a few proteins characteristically expressed by lens fiber cells in situ. Chromatin and RNA was collected from lens epithelial explants cultured in either un-supplemented media or media containing 50% bovine vitreous humor for one or five days. Chromatin for ATAC-sequencing and RNA for RNA-sequencing was prepared from explants to assess regions of accessible chromatin and to quantitatively measure gene expression, respectively. Vitreous humor increased chromatin accessibility in promoter regions of genes associated with fiber differentiation and, surprisingly, an immune response, and this was associated with increased transcript levels for these genes. In contrast, vitreous had little effect on the accessibility of the genes highly expressed in the lens epithelium despite dramatic reductions in their mRNA transcripts. An unbiased analysis of differentially accessible regions revealed an enrichment of cis-regulatory motifs for RUNX, SOX and TEAD transcription factors that may drive differential gene expression in response to vitreous.
This article was originally published in Cells. The version of record is available at: https://doi.org/10.3390/cells12030501
ATAC-seq , RNA-seq , lens , development , differentiation , explants , immune , motif , chromatin
Upreti, Anil, Stephanie L. Padula, Jared A. Tangeman, Brad D. Wagner, Michael J. O’Connell, Tycho J. Jaquish, Raye K. Palko, Courtney J. Mantz, Deepti Anand, Frank J. Lovicu, Salil A. Lachke, and Michael L. Robinson. 2023. "Lens Epithelial Explants Treated with Vitreous Humor Undergo Alterations in Chromatin Landscape with Concurrent Activation of Genes Associated with Fiber Cell Differentiation and Innate Immune Response" Cells 12, no. 3: 501. https://doi.org/10.3390/cells12030501