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Functional Investigation of TUBB4A Variants Associated with Different Clinical Phenotypes

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Abstract

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.

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Acknowledgements

We are grateful to the patients and their families for their participation in this study. We thank Prof. Jiada Li for providing cell lines and Meng Wu for technical assistance.

Funding

This work was supported by the National Natural Science Foundation of China (81771409 and 82071462 to J.P.).

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Authors and Affiliations

  1. Department of Pediatrics, Xiangya Hospital, Central South University, Xiangya Road 87, Changsha, 410005, Hunan, China

    Hui Xiao, Hailan He, Tenghui Wu, Xiaoyuan Ni, Fangyun Liu, Fei Yin & Jing Peng

  2. Hunan Intellectual and Developmental Disabilities Research Center, Changsha, 410005, China

    Jing Peng

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  1. Hui Xiao
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Contributions

Conceptualization: H.X. and J.P.; investigation: H.X., F.Y., X.N., and T.W.; resources: J.P.; writing—original draft preparation: H.X.; writing—review and editing: H.H., F.L., and J.P.

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Correspondence to Jing Peng.

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This study was approved by the Ethics Committee of Xiangya Hospital of Central South University, China (Human study/protocol #201603205) and performed in accordance with the ethical standards laid down in the Declaration of Helsinki. Written informed consent was obtained from the patients’ parents.

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Xiao, H., He, H., Wu, T. et al. Functional Investigation of TUBB4A Variants Associated with Different Clinical Phenotypes. Mol Neurobiol 59, 5056–5069 (2022). https://doi.org/10.1007/s12035-022-02900-9

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