Page 51 - Hormel Report 2023

Figure: Volcano plots illustrating differential gene expression across cell lines with distinct mutations and modifications, compared to the WT.
Figure: Heatmap illustrating the fold change in expression levels of the top 100 differentially expressed genes in different mutant cell lines, compared to the WT.
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The Role of APC Mutations in Melanoma Brain Metastasis
This project is currently funded by the Melanoma Research Alliance (Robinson) with a total award amount of $100,000. The research aims to explore the
role of dysregulated Wnt signaling in affecting the melanoma tumor immune microenvironment and metastasis.
Histone Methylation and Chromosome Instability in DIPG
This project is being funded through The Hormel Institute’s Internal Grants Program-Paint the Town Pink Award (Robinson), with an award amount of $50,000. The research focuses on understanding the effect of loss of K27 methylation on chromosomal mis-segregation in DIPG cells.
Defining the Relative Contribution of K27 Methylation and S31 Phosphorylation to Gliomagenesis in DIPG
This project has been awarded from the Department of Defense (Robinson), with a total funding amount of $556,454. The study aims to determine the specific contributions of K27 methylation loss and S31 phosphorylation to gliomagenesis in DIPG.
In summary, our lab has a diverse portfolio of both active and pending research projects that are generously funded by multiple prestigious awards. These endeavors underline our unyielding commitment to pushing the boundaries in our understanding of cancer biology.
ORCID iD: https://orcid.org/0000-0002-6585-0116
Defining the relative contribution of K27 methylation and S31 phosphorylation to gliomagenesis in DIPG.
Introduction: This study probes the complexity of gene expression in Diffuse Intrinsic Pontine Glioma (DIPG) cell lines, concentrating on mutations at Lysine 27 and Serine 31 on Histone H3.3, and the PRC2 complex knockout—key regulators in epigenetic processes. We've analyzed a spectrum of cell lines, each with distinct mutations and modifications. These were created using CRISPR HDR mutagenesis and include Wild Type (H3K27 WT), H3K27M, PRC2 Knockout in K27M, H3S31A mutation in H3K27 WT, and PRC2 Knockout in WT. Employing a range of analytical tools such as Volcano plots, Heatmaps, and Venn diagrams, we’ve discerned patterns and unique expression traits across these varied cellular conditions. Delving into the subtleties of these modifications and the ensuing epigenetic changes is crucial for unraveling
DIPG’s cellular intricacies and exploring potential epigenetic therapies.
PUBLICATION HIGHLIGHT:
• Day, C. A., Hinchcliffe, E. H., & Robinson, J. P. (2022). H3K27me3 in Diffuse Midline Glioma and Epithelial Ovarian Cancer: Opposing Epigenetic Changes Leading to the Same Poor Outcomes. Cells, 11(21),19.
Figure: Venn diagram showcasing the overlap of down-regulated genes among different cell lines compared to the WT, each exhibiting unique mutations and modifications.
Figure: Venn diagram showcasing the overlap of up- regulated genes among different cell lines compared to the WT, each exhibiting unique mutations and modifications.
Summary: Preliminary findings suggest a notable shift in gene expression in the various mutant and knockout cell lines compared to WT. The identified differentially expressed genes and their overlap provide insights into the potential biological pathways and processes affected by the mutations and knockouts.
Significance: H3.3 K27M mutations do not functionally mimic the loss of H3.3 K27 methylation. mportantly, there is a unique and shared set of genes that are regulated both by K27M mutations
and the loss of S31P phosphorylation.