中南医学科学杂志2023，Vol.51Issue(6)：797-802,6.DOI:10.15972/j.cnki.43-1509/r.2023.06.001
孤独症谱系障碍相关转录因子EB基因罕见变异导致神经元轴突发育障碍
Rare variants of transcription factor EB gene associated with autism spectrum disor-der lead to neuronal axon developmental disorder
摘要
Abstract
Aim To investigate the effects of three variants of transcription factor EB(TFEB)found in autism spectrum disorder(ASD)on neuronal axon development and the mechanisms.Methods Three TFEB plasmid mutants associated with ASD and TFEB shRNA interference plasmids were constructed.After cell transfection with the a-bove plasmids,immunofluorescence,cell nucleus-cytoplasm separation,and protein immunoblotting were used to detect whether TFEB mutations change its subcellular localization.The effect of TFEB mutations on its protein stability was de-tected by protein immunoblotting after treatment with ribosomal inhibitor CHX.Primary cultured mouse neurons were transfected with the above plasmids,and the effect of TFEB mutations on neuronal axon growth and development was detec-ted by immunofluorescence.Quantitative real-time PCR was used to detect the expression level of TFEB downstream auto-phagy-lysosome genes,and the transcriptional activity of TFEB was measured by luciferase reporter gene assay.Results ASD-related mutants p.R22Q and p.R465W increased the localization of TFEB in the cell nucleus,but the mu-tations did not change the protein's stability.Meanwhile,overexpression of wild-type(WT)TFEB,but not ASD-related TFEB mutants,increased neuronal axon length.This indicates that these mutations impair TFEB's function in promoting neuronal axon growth.At the same time,autophagy activator Rapamycin can restore the axon shortening caused by TFEB interference.Overexpression of TFEB WT increased the expression of autophagy-lysosome-related downstream target genes(LAMP1,SQSTM1,CTSB,CTSD,CTSF,MAPLC3),but ASD-related TFEB mutants reduced their regulatory ability on related target genes.Conclusion TFEB mutations may lead to autophagy-lysosome dysfunction causing abnormal neu-ronal axon development,which may be one of the mechanisms associated with ASD.关键词
孤独症谱系障碍/TFEB/自噬-溶酶体Key words
autism spectrum disorder/TFEB/autophagy-lysosome分类
生物科学引用本文复制引用
胡龙妃,谭洁琼,胡章雪..孤独症谱系障碍相关转录因子EB基因罕见变异导致神经元轴突发育障碍[J].中南医学科学杂志,2023,51(6):797-802,6.基金项目
国家科技部科技创新2030-"脑科学与类脑研究"重大项目(2021ZD0201700) （2021ZD0201700）
国家卫生健康委员会出生缺陷研究与预防重点实验室(湖南省妇幼保健院)开放课题(KF2021001) （湖南省妇幼保健院）
