We are continuing our work examining the roles of PKCθ and Wnt signaling, as well as pursuing exciting new leads emanating from our human cord blood model, in establishing/maintaining leukemia stem cell activity in T-ALL.
In the PKCθ project, we have taken a biochemical approach to dissecting downstream pathways. We developed an analog-specific kinase variant of PKCθ, followed by discovery mode mass spectrometry (LC-ESI-MS/MS) to define direct kinase targets, and are pursuing functional validation of hits. This work is being done in collaboration with Gregg Morin at BCCA.
In the Wnt project, we are utilizing our integrated, real-time fluorescent Wnt reporter construct which labels a population of Wnt-active cells that is highly enriched for leukemia-initiating cell (LIC) activity, to define gene expression programs unique to leukemia stem cells. We are also dissecting the structural elements required for activating downstream gene expression programs using conditional Lef1 and Tcf7 mice obtained in collaboration with Howard Xue at the University of Iowa. We are also starting to extend this work into our synthetic human T-ALL model and in our sizable panel of patient-derived xenografts (PDX).
In the human cord blood model, we are able to produce sufficient pre-leukemia cells to support RNA-seq, ChIP-seq, ATAC-seq, and other biochemical assays to explore molecular mechanisms contributing to establishment of leukemia stem cell activity within human committed T-lineage progenitor cells.
*This work is currently funded by a Canadian Institutes of Health Research (CIHR) Project Scheme operating grant (2016-2021).