Intermediate filament dysregulation and astrocytopathy in the human disease model of KLHL16 mutation in giant axonal neuropathy (GAN).
Intermediate filament dysregulation and astrocytopathy in the human disease model of KLHL16 mutation in giant axonal neuropathy (GAN).
复制标题
巨轴突神经病 (GAN) KLHL16 突变人类疾病模型中的中间丝失调和星形细胞病。
DOI：
10.1101/2023.03.13.532440
复制
发表时间：
2023
期刊：
影响因子：
--
通讯作者：
Snider,Natasha
中科院分区：
文献类型：
--
作者：
Battaglia,Rachel;Faridounnia,Maryam;Beltran,Adriana;Robinson,Jasmine;Kinghorn,Karina;Ezzell,JAshley;Bharucha-Goebel,Diana;Bonnemann,Carsten;Hooper,JodyE;Opal,Puneet;Bouldin,ThomasW;Armao,Diane;Snider,Natasha
巨轴突神经病 (GAN) 是一种由 KLHL16 突变引起的小儿神经退行性疾病。 KLHL16 编码 gigaxonin，一种中间丝 (IF) 蛋白周转的调节剂。之前的神经病理学研究和我们在当前研究中对死后 GAN 脑组织的检查表明星形胶质细胞参与了 GAN。为了研究潜在机制，我们将七名携带不同 KLHL16 突变的 GAN 患者的皮肤成纤维细胞重新编程为 iPSC。通过对一名携带纯合错义突变 (G332R) 的患者进行 CRISPR/Cas9 编辑，获得了具有恢复 IF 表型的同基因对照。神经祖细胞（NPC）、星形胶质细胞和脑类器官是通过定向分化产生的。所有 GAN iPSC 系都缺乏十加索尼素，但在等基因对照中却得到了恢复。与同基因对照相比，GAN iPSC 显示患者特异性波形蛋白表达增加，而 GAN NPC 巢蛋白表达减少。在 GAN iPSC 星形胶质细胞和脑类器官中观察到最引人注目的表型，它们表现出密集的核周 IF 积累和异常的核形态。具有大核周波形蛋白聚集体的 GAN 患者细胞积累了核 KLHL16 mRNA。在过表达研究中，波形蛋白存在时，GFAP 寡聚化和核周聚集得到增强。作为 KLHL16 突变的早期效应物，波形蛋白可能作为 GAN 的潜在治疗靶点。
Giant Axonal Neuropathy (GAN) is a pediatric neurodegenerative disease caused by KLHL16 mutations. KLHL16 encodes gigaxonin, a regulator of intermediate filament (IF) protein turnover. Previous neuropathological studies and our own examination of postmortem GAN brain tissue in the current study revealed astrocyte involvement in GAN. To study the underlying mechanisms, we reprogrammed skin fibroblasts from seven GAN patients carrying different KLHL16 mutations to iPSCs. Isogenic controls with restored IF phenotypes were derived via CRISPR/Cas9 editing of one patient carrying a homozygous missense mutation (G332R). Neural progenitor cells (NPCs), astrocytes, and brain organoids were generated through directed differentiation. All GAN iPSC lines were deficient for gigaxonin, which was restored in the isogenic control. GAN iPSCs displayed patient-specific increased vimentin expression, while GAN NPCs had decreased nestin expression compared to isogenic control. The most striking phenotypes were observed in GAN iPSC-astrocytes and brain organoids, which exhibited dense perinuclear IF accumulations and abnormal nuclear morphology. GAN patient cells with large perinuclear vimentin aggregates accumulated nuclear KLHL16 mRNA. In over-expression studies, GFAP oligomerization and perinuclear aggregation were potentiated in the presence of vimentin. As an early effector of KLHL16 mutations, vimentin may serve as a potential therapeutic target in GAN.