Deconstructing olfactory epithelium developmental pathways in esthesioneuroblastoma

Importance: Esthesioneuroblastoma (ENB) is a rare tumor arising from the olfactory cleft region of the nasal cavity. Due to the low incidence of this tumor, as well as an absence of established reagents such as cell lines or murine models, understanding the mechanisms driving ENB pathobiology has been challenging. Objective: Here, we sought to apply advances from research on the human olfactory epithelial neurogenic niche, along with new biocomputational approaches, to better understand the cellular and molecular factors in low and high grade ENB and how specific transcriptomic markers may predict prognosis. Design, Setting, and Participants: This was a retrospective biocomputational analysis utilizing human ENB and normal olfactory mucosal tissue and clinical outcomes data. The setting was a large academic medical center. Participants were selected based on available datasets, and included ENB patients across all four tumor grades, and patients with normal olfactory mucosa. Main Outcomes and Measures: Outcomes include deconvolution analysis, differential gene expression analysis, overall survival, and immunohistochemistry. A machine learning model was used to predict cell-type proportion signatures in ENB, and Kaplan-Meier curves with associated log-rank tests were used to estimate differences in overall survival. Results: We analyzed a total of 19 ENB samples (9 low grade I/II, 10 high grade III/IV) with available bulk RNA-Sequencing and survival data, along with 10 samples from normal olfactory mucosa (3 bulk RNA-Sequencing, 7 single cell RNA-Sequencing). The bulk RNA-Sequencing deconvolution model identified a significant increase in globose basal cell (GBC) and CD8 T cell identities in high grade tumors (GBC from approximately 0% to 8%, CD8 T cell from 0.7% to 2.2%), and significant decreases in mature neuronal, Bowmans gland, and olfactory ensheathing programs, in high grade tumors (mature neuronal from 3.7% to approximately 0%, Bowmans gland from 18.6% to 10.5%, olfactory ensheathing from 3.4% to 1.1%). Trajectory analysis identified potential regulatory pathways in proliferative ENB cells, including PRC2. Survival analysis guided by gene expression in bulk RNA-Sequencing data identified favorable prognostic markers such as SOX9, S100B, and PLP1 expression. Conclusions and Relevance: Our analyses provide a basis for additional translational research on ENB management, as well as identification of potential new prognostic markers. ### Competing Interest Statement The authors have declared no competing interest.