Supporting data for<b> </b><b>“</b><b>Activation of MAPK/AP-1 pathway mediates</b><b><i> </i></b><b><i>asxl1</i></b><b> </b><b>and</b><b><i> </i></b><b><i>IDH2</i></b>^{<i><strong>R172K</strong></i>}<b> </b><b>mutated leukemogenesis”</b>

<p dir="ltr">Co-occurring mutations of <i>asxl1</i> and <i>IDH2</i> are associated with aggressive acute myeloid leukemia (AML), yet the mechanisms underlying their synergy remain poorly understood. We generated a zebrafish model combining <i>asxl1</i> loss and <i>IDH2</i>^{<em>R172K</em>} mutation that recapitulates the clinicopathologic and molecular features of high-risk AML with differentiation blockade and reduced animal survival. The double mutant exhibited promoter hypermethylation of <i>tet2</i>, leading to its downregulation and global changes in methylation profiles. Genes pertaining to MAPK/AP-1 pathway were upregulated, leading to NADPH oxidase (NOX) expression and an increase in reactive oxygen species (ROS). Single-cell RNA-sequencing confirmed differentiation arrest in HSC-progenitor with evidence of activated ROS-MAPK/AP-1 signaling. The double mutants showed resistance to IDH2 inhibitor but were sensitive to ROS or DNA methylation targeting. <i>In silico</i> analysis of gene expression of human AML carrying co-mutation of <i>ASXL1</i>/<i>IDH2</i> also showed activation of the MAPK/AP-1 pathway. Our findings underscored an epigenetic-metabolic signaling circuit driving leukemogenesis and revealed novel therapeutic strategies for this AML subtype.</p>