THE MOLECULAR BASIS OF DIFFERENTIAL DRUG SENSITIVITY IN COLORECTAL PATIENT-DERIVED TUMOR ORGAN0IDS THE SINGLE CELL LEVEL

Abstract

It is well established that solid tumors are heterologous systems consisting of distinct clones with differential responses to drug treatment. However, screening techniques for cancer therapeutics have historically relied on isoclonal cell lines that have been in continuous culture for decades. As a consequence, information derived from these models has not always been predictive of efficacy in subsequent clinical development. More recently, tumor organoids derived from patient biopsies, with their inherent heterogeneity and presence of accessory cells, have emerged as more representative systems to study tumor biology. This study proposes to generate colorectal tumor organoids, treat them with several clinically approved drugs, and to separate out populations of cells based upon viability and transcription factor HIF-1α, which is a hypoxia marker that has been well established to regulate many of the classical tumorigenic factors seen within cancer cells. Following this, single cells will be isolated and subjected to the next-generation sequencing technique RNA-Seq to obtain mutational and gene expression profiles of the cells. Results obtained from this study have the potential to shed light on the drug sensitivity and resistance of cells within specific tumors and across colorectal cancer as a whole.