Multiciliated cells are essential for respiratory defense, using coordinated ciliary motion to expel inhaled pathogens and particles. Defects in their formation underlie chronic airway symptoms in disorders such as PCD. Although model organisms have suggested that the formation of MCCs depends on centriole amplification via the deuterosome, the mechanisms controlling this process in human cells have remained largely unknown. Using human iPS cells, the research team identified deuterosomal cells as transitional precursors that emerge early during multiciliogenesis and express hallmark genes such as CCNO, DEUP1, CDC20B, and FOXN4.

A major advancement of the study was the discovery that CD36 functions as a surface marker specifically observed on iPS cell-derived DCs. Through transcriptomic screening and fluorescence‑activated cell sorting, the team demonstrated that CD36⁺ cells are highly enriched for deuterosome-associated gene expression and exclusively give rise to multiciliated cells in vitro. This breakthrough enables, for the first time, the prospective isolation of human DCs, providing direct access to a lineage that is extremely difficult to capture in native tissues due to its brief appearance during development.

To explore how DC dysfunction contributes to disease, the researchers generated iPS cells from a pediatric patient with PCD carrying compound heterozygous loss-of-function variants in CCNO, a gene known to regulate deuterosome-mediated centriole amplification. When differentiated into airway epithelium, patient-derived cells displayed impaired multiciliogenesis, reduced expression of deuterosome-associated genes, failure to generate basal bodies, and fewer motile cilia, as confirmed by immunostaining and electron microscopy. Notably, the CD36⁺ DC population was virtually undetectable in patient cells, highlighting a failure to initiate the normal DC-to-MCC developmental pathway.

CRISPR/Cas9-based correction of the CCNO variants restored DC formation, normalized gene expression dynamics, and reestablished the capacity to generate functional multiciliated cells. Single‑cell RNA sequencing further revealed that CCNO deficiency forces cells toward aberrant, DC‑bypassing trajectories, producing immature MCC-like populations that cannot complete ciliogenesis. Corrected cells, in contrast, followed a canonical progression from common progenitors to DCs and finally to mature MCCs, closely resembling adult human airway differentiation profiles.

This work establishes a robust human iPSC-based platform for studying multiciliogenesis and uncovers the essential role of deuterosomal cells in coordinating centriole amplification and cilia formation. By enabling the isolation and functional analysis of DCs, the findings open new avenues for investigating congenital ciliopathies, modeling patient-specific disease mechanisms, and ultimately guiding the development of therapeutic strategies for airway disorders associated with defective multiciliated cell differentiation.

• doi: 10.1016/j.stemcr.2026.102860