ZFHX4 is necessary for dopaminergic neuron differentiation and controls cell cycle by regulating LIN28A

Parkinson’s disease (PD) involves selective degeneration of midbrain dopaminergic neurons (mDANs), yet the regulatory networks governing their development remain incompletely understood. ZFHX4 has been linked to neurodevelopment across species and shows reduced expression in the PD midbrain. Through integrative analysis of our multiomic data of mDAN differentiation, we show that ZFHX4 is a super-enhancer-controlled transcription factor induced during mDAN specification. Importantly, ZFHX4 is necessary but not sufficient for mDAN differentiation. Genome-wide profiling of ZFHX4 binding revealed targeting to active promoters, and transcriptomic profiling after ZFHX4 depletion identified primary target genes enriched for cell-cycle regulation. Consistently, ZFHX4-depleted cells showed reduced proliferation and accumulated in G2 phase, impairing cell-cycle progression. LIN28A, an RNA-binding protein involved in stem-cell maintenance and microRNA maturation, is among the strongest upregulated genes upon ZFHX4 depletion, with direct ZFHX4 binding at the locus. Our findings indicate that ZFHX4 regulates mDAN maturation through a mechanism involving the LIN28A-miR-9 axis.