CEP170 as a novel molecular link between centrosomal function and cerebral cortical development

The centrosome is a critical regulator of cortical development, orchestrating microtubule dynamics, cell cycle progression, and neuronal migration. Disruptions in centrosome-associated proteins have been associated with a range of neurodevelopmental disorders. CEP170, a microtubule-binding protein localized to the subdistal appendages (SDA) of centrioles, has been implicated in centrosome function, yet its role in corticogenesis remains poorly defined.

We analyzed CEP170 expression in mouse cortex using western blotting, qPCR, scRNA-seq, and spatial transcriptomics, and examined its transcript expression in the developing human cortex using scRNA-seq. Loss-of-function phenotypes were assessed via in utero electroporation of Cep170-targeting shRNAs in embryonic mouse cortex. Cell proliferation and microtubule dynamics were analyzed using CRISPR/Cas9-generated CEP170-knockout cells, flow cytometry, microtubule regrowth assays, and immunofluorescence microscopy. Protein interactions were examined via co-immunoprecipitation and subcellular localization studies.

We show that CEP170 is expressed in both neural progenitors and postmitotic neurons during cortical development. Cep170 knockdown in embryonic mouse cortex resulted in profound neuronal migration deficits, altered laminar fate, and abnormal dendritic morphology. CEP170 depletion also impaired progenitor cell proliferation both in vitro and in vivo. Mechanistically, C-terminal truncations disrupted CEP170 centrosomal and microtubule localization, mediated via impaired interactions with CCDC120. These truncations impaired microtubule regrowth and organization. Strikingly, partial deletion of CEP170's centrosomal targeting and microtubule-binding domains led to severe migration deficits in the developing cortex.

Our findings identify CEP170 as a critical regulator of neural progenitor proliferation, neuronal migration, and cortical architecture via centrosome-microtubule interactions, providing new insights into centrosome-linked neurodevelopmental disorders.