Morphogen-guided neocortical organoids recapitulate regional areal identity and model neurodevelopmental disorder pathology

Tsai, Y.-C.; Ozaki, H.; Wang, X.; Almet, A. A.; Fleming, I.; Shiraiwa, K.; Noh, M. J. M.; Nie, C.; Trejo, S.; Sugita, B. K.; Dalal, J.; Gonzalez, R. A.; De Jesus, B.; Chen, G. L.-M.; Gandal, M. J.; Nie, Q.; Watanabe, M.

doi:10.1101/2025.09.02.672952

bioRxiv · 10.1101/2025.09.02.672952

Morphogen-guided neocortical organoids recapitulate regional areal identity and model neurodevelopmental disorder pathology

Tsai, Y.-C.·Ozaki, H.·Wang, X.·Almet, A. A.·Fleming, I.·Shiraiwa, K.·Noh, M. J. M.·Nie, C.·Trejo, S.·Sugita, B. K.·Dalal, J.·Gonzalez, R. A.·De Jesus, B.·Chen, G. L.-M.·Gandal, M. J.·Nie, Q.·Watanabe, M.

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Abstract

The human neocortex exhibits characteristic regional patterning (arealization) critical for higher-order cognitive function. Disrupted arealization is strongly implicated in neurodevelopmental disorders (NDDs), but current neocortical organoid models largely fail to recapitulate this patterning, limiting mechanistic understanding. Here, we establish a straightforward method for generating arealized organoids through short-term early exposure to anterior (FGF8) or posterior (BMP4/CHIR-99021) morphogens. These treatments created distinct anterior and posterior signaling centers, supporting long-lasting polarization, which we validated with single-cell RNA sequencing that revealed area-specific molecular signatures matching prenatal human cortex. To demonstrate the utility of this platform, we modeled Fragile X Syndrome (FXS) in organoids with distinct anterior and posterior regional identities. FXS organoids showed highly disrupted SOX4/SOX11 expression gradients along the anterior-posterior axis, consistent with alterations found in autism spectrum disorder (ASD) and demonstrate how regional patterning defects may contribute to NDD pathology. Together, our study provides a robust platform for generating neocortical organoids with anterior-posterior molecular signatures and highlights the importance of modeling NDDs using experimental platforms with neuroanatomic specificity.

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Tsai, Y.-C., Ozaki, H., Wang, X., Almet, A. A., Fleming, I., Shiraiwa, K., Noh, M. J. M., Nie, C., Trejo, S., Sugita, B. K., Dalal, J., Gonzalez, R. A., De Jesus, B., Chen, G. L.-M., Gandal, M. J., Nie, Q., Watanabe, M.. 2025-09-03. Morphogen-guided neocortical organoids recapitulate regional areal identity and model neurodevelopmental disorder pathology. https://doi.org/10.1101/2025.09.02.672952

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