Characterization of human nasal organoids from chronic rhinosinusitis patients

Abstract

Patient-derived organoids grown in three-dimensional cultures provide an excellent platform for phenotypic high-throughput screening and drug response research. Organoid technology has been applied to study stem cell biology and various human pathologies. This study investigates the characteristics and cellular morphology of organoids derived from primary human nasal epithelial cells (HNECs) of chronic rhinosinusitis (CRS) patients. Nasal organoids were cultured up to 20 days and morphological, cell composition, and functional parameters were measured by immunofluorescence, RT-qPCR, western-blot and FACS analysis. The results showed that nasal organoids expressed the stem cell marker Leucine Rich Repeat Containing G Protein-Coupled Receptor 5 (LGR5), and markers for apical junction genes, goblet cells and ciliated cells. Moreover, we were able to regrow and expand the nasal organoids well after freezing and thawing. This study promise an effective and feasible method for development of human nasal organoids, suitable for the phenotypic high-throughput screening and drug response research.

Scientific Advancements That Empower Us to Understand CRS Pathophysiology

“…Most importantly, they show that organoids were able to regrow and expand after freezing and thawing to allow the banking of tissue from specialized groups of patients for drug studies in the future, an area that has historically had limitations with ALI cultures. 18 In another similar study, Franca et al demonstrate successful growth of spheroids from CRSwNP patients which were cultivated for 20 days with cells remaining differentiated for a longer time with ciliary beating compared with standard ALI cultures. 19 In summary, 3D cultures have significant potential to better dissect the cellular interactions with the sinonasal epithelium and other cell types in CRS (Table 2).…”

“…França et al 19 2021 Organoids Established that the cells derived from nasal polyp during ESS can be maintained in at three-dimensional culture environment. Ramezanpour et al 18 2022 Organoids Generated CRS patients derived nasal organoids that exhibited centralized lumen surrounded by airway epithelial cell layer, expressed Lgr5, and contained mucus-secreting and ciliated cells. 23…”

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“…The olfactory placode and olfactory sensory neurons were generated in hPSCs at a 20-30% differentiation efficiency; more research is needed to better understand the development and to increase the differentiation efficiency of the olfactory tissue [70]. Culture conditions were established for human nasal organoids [71] and for the expansion of primary human olfactory mucosa cells [72,73] which can be applied to future hPSC-differentiation and maintenance protocols. Generation of an induced hPSC line reprogrammed from human olfactory mucosa may be a potential cell line source for stem cell therapies, because this induced hPSC line may retain olfactory epigenetic signatures which may produce a more pure olfactory sensory neuron population [74].…”

Stem Cell Therapies for Sensory Organ Disorders

“…The olfactory placode and olfactory sensory neurons were generated in hPSCs at a 20-30% differentiation efficiency; more research is needed to better understand the development and to increase the differentiation efficiency of the olfactory tissue [70]. Culture conditions were established for human nasal organoids [71] and for the expansion of primary human olfactory mucosa cells [72,73] which can be applied to future hPSC-differentiation and maintenance protocols. Generation of an induced hPSC line reprogrammed from human olfactory mucosa may be a potential cell line source for stem cell therapies, because this induced hPSC line may retain olfactory epigenetic signatures which may produce a more pure olfactory sensory neuron population [74].…”