ABSTRACT Biopsy is crucial in clinical medicine to obtain tissues and cells that directly reflect the pathological changes of each disease. However, the brain is an exception due to ethical and practical challenges. Nasal biopsy, which captures the olfactory neuronal epithelium, has been considered as an alternative method of obtaining neuronal cells from living patients. Multiple groups have enriched olfactory neuronal cells (ONCs) from biopsied nasal tissue. ONCs can be obtained from repeated biopsies in a longitudinal study, providing mechanistic insight associated with dynamic changes along the disease trajectory and treatment response. Nevertheless, molecular characterization of biopsied nasal cells/tissue has been insufficient. Taking advantage of recent advances in next-generation sequencing technologies at the single-cell resolution and related rich public databases, we aimed to define the neuronal characteristics, homogeneity, and utility of ONCs. We applied single-cell and bulk RNA sequencing for ONCs, analyzing and comparing the data with multiple public datasets. We observed that the molecular signatures of ONCs are similar to those of neurons, distinct from major glial cells. The signatures of ONCs resemble those of developing neurons and share features of excitatory neurons in the prefrontal and cingulate cortex. The high homogeneity of ONCs is advantageous in pharmacological, functional, and protein studies. Accordingly, we provide two proof-of-concept examples for functional and protein studies, solidifying the utility of ONCs in studying objective biomarkers and molecular mechanisms for brain disorders. The ONCs may also be useful in the studies for the olfactory epithelium impairment and the resultant mental dysfunction elicited by SARS-CoV-2. SIGNIFICANCE STATEMENT To study dynamic changes and underlying mechanisms along disease trajectory and treatment response in neuropsychiatric disorders, olfactory neuronal cells (ONCs) enriched from biopsied nasal tissue may provide a crucial tool. Because ONCs can be obtained from repeated biopsies in a longitudinal study, this tool has been believed to be useful and complementary to postmortem brains and induced pluripotent stem cell-derived neurons. Nevertheless, molecular characterization of biopsied nasal cells/tissue has been insufficient, which hampers a broader use of this resource. Taking advantage of recent advances in next-generation sequencing technologies at the single-cell resolution and related rich public databases, the present study defines ONCs’ neuronal characteristics, homogeneity, and unique utility for the first time.

Human olfactory neuronal cells through nasal biopsy: molecular characterization and utility in brain science