显性TUBB4A变异导致不同的表型,包括伴有基底节和小脑萎缩的髓鞘减少(H-ABC),肌张力障碍4型(DYT4),和孤立的髓鞘减少。这里,我们报告了4例新的TUBB4A变体患者(p。K324T)和三名先前报道的变异的新患者(p。Q292K,p.V255I,p.E410K)。携带新型p.K324T变体的个体表现出婴儿期癫痫伴转移性局灶性癫痫发作(EIMFS),而其他三个具有分离的髓鞘减少表型。我们还对负责H-ABC的TUBB4A变体的细胞作用进行了研究(p。D249N),DYT4(第R2G),严重的联合表型,结合了髓鞘减少和EIMFS(p。K324T),和孤立的髓鞘减少(p。Q292K和p.E410K)关于微管稳定性和动力学,神经突生长,树突脊柱发育,和驱动蛋白结合。基于细胞的测定显示,除p.R2G外,所有变体都会增加微管稳定性,降低微管聚合速率,减少轴突生长,并改变树突棘的密度和形状。我们还发现p.K324T和p.E410K变体扰乱了TUBB4A与KIF1A的结合,突触小泡前体运输所需的神经元特异性驱动蛋白。一起来看,我们的数据表明,微管稳定性和动力学受损,轴突生长缺陷,树突脊柱发育是TUBB4A相关脑白质营养不良的共同分子基础。TUBB4A与KIF1A的结合受损更可能参与分离的髓鞘减少表型,这表明驱动蛋白结合的改变可能导致不同的表型。总之,我们的研究扩展了TUBB4A突变和相关表型的范围,并深入了解了为什么不同的TUBB4A变异导致不同的临床表型.
Dominant
TUBB4A variants result in different phenotypes, including hypomyelination with atrophy of the basal ganglia and cerebellum (H-ABC), dystonia type 4 (DYT4), and isolated hypomyelination. Here, we report four new patients with a novel
TUBB4A variant (p.K324T) and three new patients with previously reported variants (p.Q292K, p.V255I, p.E410K). The individual carrying the novel p.K324T variant exhibits epilepsy of infancy with migrating focal seizures (EIMFS), while the other three have isolated hypomyelination phenotype. We also present a study of the cellular effects of
TUBB4A variants responsible for H-ABC (p.D249N), DYT4 (p.R2G), a severe combined phenotype with combination of hypomyelination and EIMFS (p.K324T), and isolated hypomyelination (p.Q292K and p.E410K) on microtubule stability and dynamics, neurite outgrowth, dendritic spine development, and kinesin binding. Cellular-based assays reveal that all variants except p.R2G increase microtubule stability, decrease microtubule polymerization rates, reduce axonal outgrowth, and alter the density and shape of dendritic spines. We also find that the p.K324T and p.E410K variants perturb the binding of TUBB4A to KIF1A, a neuron-specific kinesin required for transport of synaptic vesicle precursors. Taken together, our data suggest that impaired microtubule stability and dynamics, defected axonal growth, and dendritic spine development form the common molecular basis of TUBB4A-related leukodystrophy. Impairment of TUBB4A binding to KIF1A is more likely to be involved in the isolated hypomyelination phenotype, which suggests that alterations in kinesin binding may cause different phenotypes. In conclusion, our study extends the spectrum of
TUBB4A mutations and related phenotypes and provides insight into why different
TUBB4A variants cause distinct clinical phenotypes.