TMIC-63. SINGLE CELL SEQUENCING OF RECURRENT GLIOBLASTOMA DEFINES CELLULAR STATES ASSOCIATED WITH THERAPY RESISTANCE

Abstract

Abstract Glioblastoma (GBM), an aggressive, primary tumor is composed of multiple clonal populations. Resistance to therapy is a hallmark of GBM, making it a deadly disease. Upon therapeutic stress, tumor heterogeneity and glial stem cell plasticity within the primary tumor contributes to reprogramming of the transcriptional activity. This alters the tumor landscape and generates therapy resistant clonal populations. Single cell RNA sequencing studies of primary GBM have defined various tumor cell states namely, the NPC-like (Neuronal precursor-like), OPC-like (oligodendrocyte precursor-like) and MES-like (mesenchymal-like) cellular states. These studies have also defined the immune components and the normal brain cells associated with primary GBM. Yet, there are no studies focused on the single cell analysis of recurrent GBM. To define the tumor landscape at recurrence, we performed single cell RNA sequencing of 180K cells from excised tumor tissue obtained from 11 patients after failure of therapeutic interventions. A corresponding cohort of 240K cells were sequenced from primary tumors obtained from 13 patients. We performed cellular state analyses of tumor cells in both the primary (~70K) and recurrent (~10K) samples. Primary GBM showed higher fractions of AC-like (0.55), compared to NPC-like (0.10), OPC-like (0.20) and MES-like (0.15). On the other hand, recurrent tissues have very low AC-like (0.09) but elevated NPC-like (0.23), OPC-like (0.40) and MES-like (0.27). This data shows a significant decrease in AC-like cells and corresponding increase in MES-like, OPC-like, NPC-like cells after recurrence. Within this dataset we also have matched longitudinal samples for analysis. We are currently performing trajectory analysis to identify genes and transcription programs activated during tumor recurrence. Experimentally, using known markers for the different cellular states we have also performed immunofluorescence analyses to validate the different cell states altered during recurrence. Our efforts to generate comprehensive recurrent GBM atlas will prove to be valuable to the GBM community.