Establishment of a Patient-Derived Organoid Culture Model of Olfactory Neuroblastoma

Olfactory neuroblastoma (ONB) is a rare malignant tumor arising from the olfactory epithelium. Current therapeutic approaches have not significantly changed over the previous decades, and two-year survival of high grade ONB remains low at approximately 40%, partially due to an incomplete understanding of tumor biology. An absence of robust pre-clinical or culture models has been a barrier for the development of new treatment strategies. Here we sought to establish an in vitro patient-derived culture model of ONB in order to allow for downstream targeted manipulation of tumor biology and therapeutic testing. Tumor samples from two subjects with biopsy-confirmed ONB were used to explore in vitro culture development, in accordance with IRB protocols. Given evidence for cellular heterogeneity within ONB, we tested an organoid growth model utilizing ultra-low adherent plates with the goal of maintaining 3D architecture within tumors. Successful cultures were assessed immunochemically, and compared to parent tumor immunohistochemistry. We successfully established ONB organoids derived from two Hyam’s grade II tumors. After 2-3 weeks in culture, organoids developed as round spheres and could be passaged for >5 months. Tumor organoid immunofluorescence confirmed expected olfactory lineage marker expression. Furthermore, similar cellular heterogeneity to that observed in the original tumor samples was maintained in organoids, defined by olfactory epithelial basal, neural and nonneuronal lineage factors. To our knowledge, our cultures represent the first in vitro model of human ONB that successfully recapitulates in vivo tumor heterogeneity, and they may facilitate drug screens and experimental manipulation not previously possible in ONB research.

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Artikel online veröffentlicht:
05. Februar 2024