The Study of NSD2 Biochemical and Biological Activity Through Small-Molecule Profiling of Cell Lines

Abstract

Histone methyltransferase NSD2 (WHSC1/MMSET) is correlated with several cancers, but promising NSD2 inhibitors are yet to be developed for the clinic, alternative strategies should be studied parallel to probe development in targeting NSD2-driven cancer. In this thesis, I employ traditional biochemical, cellular biology methods and small molecule profiling to study a recurrent activating mutation of NSD2, E1099K. This study sought identify orthogonal proteins that can be targeted to inhibit the growth of E1099K-NSD2 cancers. The body of work describes biochemical and cellular approaches in studying E1099K-NSD2 that revealed E1099K-NSD2 to mono- and di- methylate NSD2 native substrate of H3K36, leading to transcriptional signatures similar to cell lines with overexpression of NSD2. To identify NSD2 E1099K-induced dependencies of cell lines, a panel of hematopoietic cancer cell lines, were profiled against a set of small molecules to establish cell viability in response to treatment. HDAC 1/2 inhibitors, were identified to have greater potency for killing E1099K- compared to WT-NSD2 cell lines. CRISPR technology was applied to engineer isogenic cell lines with different NSD2 genomic status and profiled them against the set of small-molecules. RNA-seq identified oncogenic pathway and PRC2 targets to be upregulated in E1099K-NSD2 clones. Further analysis of clones revealed a small molecule study on mycoplasma colonized cells that resulted in a brief study descibed at the end of the thesis.