Mentor Name: Brandon Hadland

This project focuses on CBFA2T3-GLIS2–driven acute megakaryoblastic leukemia (AMKL), one of the most aggressive forms of childhood leukemia. This leukemia originates before birth, when a chromosomal inversion creates the CBFA2T3-GLIS2 fusion and establishes a pre-leukemic state. Although children may remain healthy for months to years after birth, this early lesion can later evolve into full leukemia. Even after treatment, leukemia-initiating stem cells (LSCs) often survive by entering a dormant state, making them highly resistant to therapy and contributing to relapse. Despite their clinical importance, the processes that allow LSCs to emerge and persist remain poorly understood. To investigate these mechanisms, we use a CRISPR/Cas9-based approach to precisely recreate the natural chromosome 16 inversion that forms the CBFA2T3-GLIS2 fusion in human cord blood and pluripotent stem cell–derived blood progenitors. We pair this system with an endothelial coculture model that mimics the supportive microenvironment that protects LSCs from chemotherapy. This platform enables us to study how this leukemia develops, how LSCs interact with their surrounding niche, and why these interactions promote resistance. Building on our prior work identifying niche-derived signals that help LSCs survive treatment, this project will test targeted strategies to disrupt these survival pathways and increase LSC sensitivity to therapy. By integrating advanced leukemia modeling with single-cell transcriptomics, lineage tracing, and computational analyses, we aim to uncover therapeutic vulnerabilities in LSCs and create a pipeline to translate these findings into future treatments. During the summer, the ALSF POST intern will lead experiments evaluating combinations of inhibitors predicted to target key pathways required for LSC survival. These studies will use our established engineered leukemia models to identify synergistic therapeutic strategies with potential to guide future therapy development for children with AMKL.