Project Summary/Abstract Candidate: I am a postdoctoral fellow in the lab of Dr. Ross Levine in the Human Oncology & Pathogenesis Program at Memorial Sloan Kettering Cancer Center (MSKCC). My PhD studies allowed me to hone the technical and experimental skills required for interrogating clinically tractable molecular dependencies in cancer cells. My current research focuses on the generation of models of acute myeloid leukemia (AML) evolution to be used in the discovery of novel molecular dependencies and potential therapeutic targets. To that end, I have broadened my capabilities with CRISPR editing to generate an inducible AML model that allows for temporal control of mutagenesis and optimized a single cell DNA sequencing technique to evaluate the clonal framework of AML. My proposed research will build upon these initial studies to develop a new suite of sequential mutagenesis models of AML. These models developed in the K99 phase of this grant will serve as tools for the discovery of essential proteins/pathways for leukemic cells. My long-term career goal is to lead an independent research group focused on the identification and characterization of molecular dependencies of AML using precise models of disease evolution through sequential mutagenesis. To accomplish these goals, I have outlined a career plan that will 1) expand of my technical skills and scientific capacity, 2) improve my scientific communications with the field, 3) advance my supervisory and leadership abilities, 4) develop and foster collaborative relationships and 5) prepare me for the transition to independence. Project: Molecular profiling studies of AML patients infer a progressive acquisition of mutations that drives leukemogenesis, but are unable to delineate the dominant clonal framework leading to disease or identify the precise mutational order for certain genes, such as NPM1. Current models of AML are unable to truly recapitulate the step-wise mutagenesis observed in patients. Our single cell sequencing studies have further resolved the clonal structure of AML at single cell resolution and with these studies, I aim to generate new models that accurately depict the sequential mutagenesis of AML evolution. The specific aims are: 1) examine mechanisms of co-mutational clonal dominance and mutation order in AML patients using single cell profiling, 2) determine the impact of mutational acquisition on disease development and progression of NPM1-mutant AML, and 3) elucidate molecular dependencies of disease derived from mutant NPM1 and co-occurring mutations. Environment: The Levine lab is part of the MSKCC Molecular Cancer Medicine Service, Human Oncology & Pathogenesis Program (HOPP), for which Dr. Levine is Chief. The Levine lab is a core member of the Center for Hematologic Malignancies and the Center for Epigenetics Research, directed by Dr. Abdel-Wahab and Dr. Kristian Helin, respectively. These affiliations at MSKCC, a state-of-the-art institution, provide a rich set of collaborative, technical and scientific resources to perform the research and career development proposed here.

Project Narrative While genetic profiling studies have proposed that a step-wise acquisition of mutations drives acute myeloid leukemia (AML) development, the impact of this mutation order on leukemogenesis has not been fully delineated. The proposal described herein aims to interrogate the clonal architecture of AML, particularly NPM1-mutant AML, at the single cell level to identify precise mutation order and common co-occurring mutational patterns which lead to clonal dominance for the development of novel sequential mutagenesis models of AML. These studies will generate accurate models of leukemogenesis to uncover molecular dependencies as clinically tractable targets of NPM1-mutant AML.