Dissecting the role of IER3IP1 in neurogenesis and brain malformation

剖析 IER3IP1 在神经发生和脑畸形中的作用

• 批准号: 10672753
• 负责人: Lucie Yeongran Ahn
• 金额: $ 5.27万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-01 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10672753
• 关键词: 3-Dimensional; Affect; Age; Biogenesis; Birth; Brain; CRISPR/Cas technology; Calnexin; Cell Death; Cell Line; Cell Proliferation; Cell model; Cell physiology; Cell surface; Cells; Centrioles; Cerebral cortex; Cerebrum; Child; Childhood; Clinical; Congenital Abnormality; Cytokinesis; DNA Damage; Data; Defect; Development; Disease model; Endoplasmic Reticulum; Gender; Generalized Epilepsy; Genes; Genetic; Golgi Apparatus; Head circumference; Hela Cells; Homologous Gene; Human; In Vitro; Knock-out; Link; Membrane; Microcephaly; Mitosis; Molecular; Mutation; Neural tube; Neurodevelopmental Disorder; Neuronal Differentiation; Neuronal Dysfunction; Neurons; Online Mendelian Inheritance In Man; Organoids; Pathogenesis; Pathogenicity; Pathway interactions; Patients; Pattern; Phenocopy; Phenotype; Population; Process; Protein Biosynthesis; Proteomics; Rare Diseases; Reporter; Reporting; Research; Role; SHH gene; Scientist; Specific qualifier value; Structural Congenital Anomalies; Syndrome; Technology; Testing; United States; Variant; Vesicle; Work; Yeasts; anterograde transport; brain abnormalities; brain malformation; cell type; cerebral atrophy; comparison control; exome sequencing; gene correction; genetic variant; genome sequencing; induced pluripotent stem cell; inhibitory neuron; insight; knock-down; morphogens; neonatal diabetes mellitus; nerve stem cell; neural; neurodevelopment; neurogenesis; neuron development; neuroregulation; patched protein; protein expression; protein transport; proteostasis; receptor; response; secretory protein; sonic hedgehog receptor; stem cells; tool; trafficking

Project Summary Primary microcephaly is a structural birth defect characterized by a significantly smaller head circumference than the mean of age and gender at birth. It is mainly linked to dysfunction of the neural progenitor cell population, resulting in reduced neurons in the cerebral cortex. Most microcephaly-causing genes are involved in mitosis and cytokinesis; however, recently, defects in secretory protein trafficking have been implicated in causing microcephaly. One of the newly identified genes that are potentially involved in protein trafficking is IER3IP1. Bi-allelic variants of IER3IP1 have been reported to cause a rare, pediatric, neurodevelopmental condition called microcephaly with simplified gyration, generalized epilepsy, and permanent neonatal diabetes syndrome (MEDS). MEDS patients present with severe congenital structural brain abnormalities, including microcephaly, ventriculomegaly, and cerebral atrophy. Prior studies on IER3IP1 suggest that its function is involved in ER to the Golgi trafficking in humans. I confirmed that the pathogenic variant blocks ER to Golgi trafficking with a GFP trafficking reporter, and the correction of IER3IP1 mutation through CRISPR/Cas9 restored the trafficking efficiency. Herein, I propose to focus on elucidating the function of IER3IP1 in neurogenesis and human brain development to understand how faulty protein trafficking leads to brain birth defects. I hypothesize that IER3IP1 regulates secretory protein trafficking during neurogenesis. I have successfully generated and characterized cortical brain organoids and 2D neuronal culture derived from MEDS patient-derived induced pluripotent stem cells and isogenic control cells to test this. Using these MEDS patient cell models, I will complete the following aims: Aim 1. What is the underlying cellular mechanism of microcephaly seen in MEDS patients with IER3IP1 p.L78P mutation? and Aim 2. Does IER3IP1 regulate the molecular trafficking of membrane receptors important in neurodevelopment? This proposal will fill the gap in our understanding of how protein trafficking regulates neurogenesis and provide insight into how abnormal protein trafficking leads to structural brain abnormalities.

项目摘要 原发性小头畸形是一种结构性出生缺陷，其特征是头部明显较小 出生时年龄和性别的平均值。它主要与神经祖细胞的功能障碍有关 细胞群，导致大脑皮层神经元减少。大多数导致小头畸形的基因 参与有丝分裂和胞质分裂;然而，最近，分泌蛋白运输的缺陷已经被发现。 与导致小头畸形有关一个新发现的基因可能与蛋白质 这是IER 3 IP 1。据报道，IER 3 IP 1的双等位基因变体引起罕见的儿科， 神经发育状况，称为小头畸形伴简单旋转，全身性癫痫，和永久性 新生儿糖尿病综合征（MEDS）。MEDS患者存在严重的先天性结构脑 畸形，包括小头畸形、脑室扩大和脑萎缩。先前对IER 3 IP 1的研究表明， 它的功能是参与ER向高尔基体的人口贩卖。我证实了致病变异体 ER到高尔基体的运输与GFP运输报告基因，以及IER 3 IP 1突变的校正， CRISPR/Cas9恢复了贩运效率。在此，我建议重点阐明IER 3 IP 1的功能 在神经发生和人类大脑发育中，了解错误的蛋白质运输如何导致大脑出生 缺陷 我推测，IER 3 IP 1调节神经发生过程中分泌蛋白的运输。我有 成功生成并表征了源自MEDS的皮质脑类器官和2D神经元培养物 患者来源的诱导多能干细胞和同基因对照细胞来测试这一点。使用这些药物的患者 细胞模型，我将完成以下目标：目标1。什么是潜在的细胞机制的小头畸形 在IER 3 IP 1 p.L78P突变的MEDS患者中观察到的？目标2 IER 3 IP 1是否调节 膜受体在神经发育中的重要作用？这项建议将填补我们在这方面的差距。 了解蛋白质运输如何调节神经发生，并深入了解异常蛋白质 贩卖会导致大脑结构异常