3' sequencing from human Huntington''s disease and control motor cortex and cerebellum.

Purpose: We applied polyA site sequencing (Passeq) to human Huntington''s disease and control motor cortex and cerebellum to test if any genes change 3'UTR isoforms abundance. Methods: 3' sequencing was performed on 6 motor cortices from grade 1 Huntington''s patient brains, 4 motor cortices from grade 2 Huntington''s patient brains, and 5 motor cortices from control brains. Cerebellum samples included 9 cerebella from grade 2-3 Huntington''s patient brains, and 7 cerebella from control brains. To verify HTT isoforms in mice, sequencing was performed on 5 Q140 mouse striata and 3 wild-type mouse striata. Results: We report 11% of genes from Huntington''s disease patient motor cortex exhibit a change in at least one of their 3'UTR isoforms, commensurate with the 11% of genes which show different total expression in HD motor cortex versus control. In contrast, gene isoform and expression changes are minimal (<5%) in Huntington''s disease cerebellum versus controls. In the motor cortex, we show isoform and gene expression differs between between grade 1 and grade 2 brains. We identify a novel isoform of huntingtin mRNA which is conserved in wild-type and Huntington''s model mice. Conclusions: This is the first study characterizing widespread alterations in 3'UTR isoform abundance in Huntington''s disease. Alterations in isoform abundance may affect mRNA metabolism in Huntington''s disease brains. Overall design: Examination of 3'UTR isoform abundance in Huntington''s disease and control motor cortex and cerebellum, and in Q140 and wild-type mouse striatum. 3' sequencing was performed on 6 motor cortices from grade 1 Huntington''s patient brains, 4 motor cortices from grade 2 Huntington''s patient brains, and 5 motor cortices from control brains. Cerebellum samples included 9 cerebella from grade 2-3 Huntington''s patient brains, and 7 cerebella from control brains. To verify HTT isoforms in mice, sequencing was performed on 5 Q140 mouse striata and 3 wild-type mouse striata.

研究目的：我们将polyA位点测序（polyA site sequencing, Passeq）应用于人类亨廷顿病患者及对照的运动皮层与小脑组织，以检测是否存在基因的3'非翻译区（3' untranslated region, 3'UTR）异构体丰度发生改变的情况。研究方法：本研究对6例1级亨廷顿病患者大脑运动皮层、4例2级亨廷顿病患者大脑运动皮层以及5例对照大脑运动皮层开展3'端测序。小脑样本涵盖9例2-3级亨廷顿病患者的小脑组织与7例对照小脑组织。为验证小鼠体内亨廷顿基因（HTT）异构体的表达情况，我们对5例Q140小鼠纹状体与3例野生型小鼠纹状体进行了测序。研究结果：我们发现，亨廷顿病患者运动皮层中11%的基因至少存在一种3'UTR异构体的丰度改变，这一比例与亨廷顿病（Huntington's disease, HD）运动皮层中总表达量存在差异的基因占比（11%）相符。与之形成鲜明对比的是，亨廷顿病患者小脑组织的基因异构体与表达量变化幅度极小（<5%），与对照组相比无显著差异。在运动皮层中，我们观察到1级与2级亨廷顿病患者大脑的基因异构体及表达量存在显著差异。此外，我们鉴定出一种亨廷顿mRNA的新型异构体，该异构体在野生型小鼠与亨廷顿病模型小鼠中均具有保守性。研究结论：本研究是首个系统性表征亨廷顿病中3'UTR异构体丰度广泛改变的研究。异构体丰度的改变可能会影响亨廷顿病患者大脑内的mRNA代谢过程。整体实验设计：探究亨廷顿病患者及对照的运动皮层、小脑组织，以及Q140小鼠与野生型小鼠纹状体中的3'UTR异构体丰度。本研究对6例1级亨廷顿病患者大脑运动皮层、4例2级亨廷顿病患者大脑运动皮层以及5例对照大脑运动皮层开展3'端测序。小脑样本涵盖9例2-3级亨廷顿病患者的小脑组织与7例对照小脑组织。为验证小鼠体内HTT异构体的表达情况，我们对5例Q140小鼠纹状体与3例野生型小鼠纹状体进行了测序。