This project will explore the differences between DNA methylation, a process by which the DNA building blocks are modified, in cancer cells and in healthy cells. DNA methylation affects gene expression and plays a key role in tumorigenesis. By analyzing many chronic lymphocytic leukemia (CLL) patient samples, our group has uncovered pervasive disordered methylation: high variability of DNA methylation across diverse regions of DNA. This is not seen in cells from healthy people. We have previously shown that higher methylation variability in CLL cells is associated with poorer clinical outcomes, suggesting that a better understanding of this phenomenon may lead to new clinical interventions. Interestingly, we also observed that these changes are present early in cancer development, suggesting that disordered methylation may be a central driver leading to cancer development. These findings provide the foundation for my research, which aims to decipher the impact of disordered methylation on the development, onset, and severity of CLL.
To pursue these aims, I will employ new mouse models of CLL reflecting key CLL driver mutations first identified in our lab. I will also utilize state-of-the-art technologies such as single-cell RNA-sequencing and single-cell methylation profiling, to determine the chronologic origin of disordered methylation with respect to cancer diagnosis and progression. I have demonstrated that mouse CLL cells, like their human counterpart, present with disordered methylation. This provides an exciting and amenable disease context in which I can monitor mice over the course of disease. Furthermore, I aim to elucidate the cellular pathways affecting disordered methylation. Using CRISPR-mediated gene-editing tools, I will target genes that might affect disordered methylation, such as the DNMT3 and TET families, and monitor the development of CLL. Results from these studies will enhance our growing understanding of CLL development as well as the development of other cancers that demonstrate disordered methylation. Ultimately, our findings may inform the development of new tools to predict cancer development in healthy individuals and identify new therapeutic targets for blood cancer patients.