Elucidating ADNP Syndrome Gene Dysregulation through a Novel Approach in iPSC-Derived GABAergic Neuron Development

Traditional approaches for differentiating human pluripotent stem cells (PSCs) into neuronal cells are often hindered by challenges such as incomplete neural differentiation, prolonged culture times, and variability across PSC lines. To overcome these limitations, we developed an innovative strategy by combining the overexpression of transcription factors Ascl1 and Dlx2 with dual-SMAD and WNT inhibition, effectively driving the differentiation of PSCs into diverse, region-specific GABAergic neuron subtypes.

Through single-cell sequencing, we thoroughly characterized the heterogeneity of these induced neurons (iNs) and uncovered the regulatory mechanisms underlying their fate specification. Our findings demonstrate the successful generation of multiple disease-relevant GABAergic neuron types. Furthermore, we investigated the impact of the ADNP syndrome-associated p.Tyr719* variant on GABAergic neuron differentiation, revealing that this mutation significantly alters neuronal subtype composition and synaptic transmission.