Multidimensional Functional Profiling of Human Neuropathogenic Alleles in Primary Cultures of Murine Pallial Precursors.

is an ancient transcription factor gene mastering telencephalic development. A number of distinct structural mutations lead to the " syndrome", a complex and heterogeneous neuropathological entity, for which no cure is presently available. Reconstruction of primary neurodevelopmental/physiological anomalies evoked by these mutations is an obvious pre-requisite for future, precision therapy of such syndrome. Here, as a proof-of-principle, we functionally scored three neuropathogenic alleles, , , and , against their healthy counterpart. Specifically, we delivered transgenes encoding for them to dedicated preparations of murine pallial precursors and quantified their impact on selected neurodevelopmental and physiological processes mastered by Foxg1: pallial stem cell fate choice, proliferation of neural committed progenitors, neuronal architecture, neuronal activity, and their molecular correlates. Briefly, we found that and generally performed as a gain- and a loss-of-function-allele, respectively, while acted as a mild loss-of-function-allele or phenocopied . These results provide valuable hints about processes misregulated in patients heterozygous for these mutations, to be re-addressed more stringently in patient iPSC-derivative neuro-organoids. Moreover, they suggest that murine pallial cultures may be employed for fast multidimensional profiling of novel, human neuropathogenic alleles, namely a step propedeutic to timely delivery of therapeutic precision treatments.