Dr. Tamilla Nechiporuk's proposed work will be performed in the laboratory of Dr. Jeffrey Tyner, Unit Director, at the OHSU Knight Cancer Institute. Under Dr. Tyner's leadership, the Unit recently completed and publicly released a first wave of Beat AML data, representing a multifaceted collection of information on drug sensitivities and genomic profiles of 900+ patients with acute myeloid leukemia (AML) and other hematologic malignancies. The Beat AML project is a multi-center consortium focused on advancing new molecular targeted drug therapies for the treatment of AML into clinical trials. More recently, with significant contributions from Dr. Nechiporuk, the Unit began to address a continuous problem of drug resistance, either acquired in relapsed disease or as an up- front, intrinsic property, which hinders the success of individualized therapies. To propel our mechanistic knowledge of molecular underpinnings of drug insensitivities and corresponding networks of signaling, Dr. Nechiporuk is leading a group utilizing non-biased genome-wide CRISPR Cas9 perturbations in AML cell lines and primary patient cells. To accomplish this objective, Dr. Nechiporuk proposes to interrogate parental sensitive and drug-exposure-rendered resistant AML cells for loss and gain of sensitivities in a variety of cell lines representing the diverse genetic background of AML. The goal of the proposed work is 1) to create a library of data informing mechanisms leading to the development of drug resistance in specific genetic backgrounds, 2) to expose potential combinatorial or alternative treatments to prevent or combat the development of resistance, and 3) to reveal unrealized novel drug targets. As laid out in the proposal, Dr. Nechiporuk plans to achieve these directives by carrying out genome-wide CRISPR Cas9 perturbations targeting patient-derived AML cells exposed to current cutting edge molecular agents. This approach will reveal mechanisms of drug resistance and unrealized molecular targets and/or agents to overcome drug resistance and will be significantly aided by implementing innovated orthogonal CRISPRi/a methodology to streamline and accelerate validations of top hits. The existing Beat AML dataset and CRISPR-revealed gene essentiality in primary patient cells will further inform CRISPR nominated pathways/targets and molecular agents for clinical relevance and application. Collectively, the work proposed here will be a major benefit to the Unit, and will facilitate numerous avenues of mechanistic validation in the laboratory as well as clinical development of novel therapeutic approaches.
This proposal is to support Dr. Nechiporuk in her ongoing efforts to implement genome-wide functional screening in acute myeloid leukemia (AML) and related malignancies, to discover genetic dependencies underlying drug sensitivity and to identify new combinatorial therapies. The proposed work centers on genome-wide orthogonal CRISPR Cas9 perturbations in drug-sensitive and resistant AML cell lines and will generate a library of data regarding mechanistic underpinnings of drug resistance and sensitivity. Additionally, this work will reveal new combinatorial treatment strategies to prevent or combat resistance and provide insights into novel drug dependencies.
