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

基本信息

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
项目类别：
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

项目摘要

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.

项目概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周长高于出生时年龄和性别的平均值。主要与神经祖细胞功能障碍有关细胞数量，导致大脑皮层神经元减少。大多数引起小头畸形的基因是参与有丝分裂和胞质分裂；然而，最近，分泌性蛋白质运输的缺陷已被发现与引起小头畸形有关。新发现的可能与蛋白质相关的基因之一贩运是 IER3IP1。据报道，IER3IP1 的双等位基因变异会导致罕见的儿科、称为小头畸形的神经发育疾病，伴有简单回转、全身性癫痫和永久性癫痫新生儿糖尿病综合征（MEDS）。 MEDS 患者患有严重的先天性大脑结构畸形，包括小头畸形、脑室扩大和脑萎缩。先前对 IER3IP1 的研究表明它的功能涉及人类的 ER 到高尔基体的贩运。我确认致病变异会阻断使用 GFP 运输记者将 ER 运输至高尔基体，并通过以下方式纠正 IER3IP1 突变 CRISPR/Cas9恢复了运输效率。在此，我建议重点阐述IER3IP1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诞生缺陷。我假设 IER3IP1 在神经发生过程中调节分泌蛋白运输。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对此进行了测试。使用这些 MEDS 患者细胞模型，我将完成以下目标：目标1. 小头畸形的潜在细胞机制是什么是否见于具有 IER3IP1 p.L78P 突变的 MEDS 患者？目标 2. IER3IP1 是否调节分子膜受体的运输对神经发育很重要？该提案将填补我们的空白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如何贩运导致大脑结构异常。