Deletion of TUBB4A mitigates the oligodendrocyte and neuronal deficits in human iPSCs derived from individuals affected by H-ABC

TUBB4A-related leukodystrophy (TUBB4A-LD) is a rare neurologic disorder with a broad spectrum of phenotypes, including severe early infantile encephalopathy, late infantile Hypomyelination with Atrophy of the Basal ganglia and Cerebellum (H-ABC), and milder late infantile forms. H-ABC is closely associated with a recurrent pathogenic variant, p.Asp249Asn, in the gene encoding tubulin beta class IVA (TUBB4A), a microtubule component. H-ABC presents with progressive dystonia, mobility loss, aphasia, and swallowing dysfunction in childhood. H-ABC results in cell-autonomous deficits in oligodendrocytes (OLs), cerebellar granule neurons, and medium spiny neurons (MSNs). Antisense oligonucleotides targeting Tubb4a can alleviate symptoms in H-ABC mouse models. However, the efficacy and safety of TUBB4A knockout in human cells remain poorly understood. We studied patient-derived TUBB4AD249N, TUBB4A KO, and control individual pluripotent stem cells (iPSCs). TUBB4AD249N iPSC-derived OLs failed to mature, showing less complexity and myelination, reduced microtubule acetylation and detyrosination. TUBB4AD249N iPSC-derived MSNs also showed impaired maturation and neurite extension. TUBB4A KO in mutant iPSCs reduced cellular deficits and was well tolerated. These findings support that suppression of TUBB4A could be a safe, effective therapy for TUBB4A-LD.