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.

项目摘要原发性小头畸形是一种结构性出生缺陷，其头部明显较小周长比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种群，导致脑皮质中的神经元减少。大多数引起微畸形的基因是参与有丝分裂和细胞因子；但是，最近，分泌蛋白运输的缺陷已经与引起小头畸形有关。潜在参与蛋白质的新鉴定的基因之一贩运是IER3IP1。据报道，IER3IP1的Bi-Callelic变体会引起罕见的小儿，神经发育状况称为小头畸形，具有简化的回旋，广义癫痫和永久性新生儿糖尿病综合征（MEDS）。药物患者出现严重的先天性结构大脑异常，包括小头畸形，心室肿瘤和脑萎缩。关于IER3IP1的先前研究表明它的功能与ER有关，与人类的高尔基人贩运有关。我证实了病原变体块与GFP贩运记者一起进行高尔基人贩运，以及通过 CRISPR/CAS9恢复了贩运效率。在此，我建议专注于阐明IER3IP1的功能在神经发生和人脑发育中，了解蛋白质运输的错误如何导致脑出生缺陷。我假设IER3IP1在神经发生过程中调节分泌蛋白的运输。我有成功产生和表征的皮质脑器官和衍生自药物的2D神经元培养患者衍生的诱导多能干细胞和等源性对照细胞进行了测试。使用这些药物细胞模型，我将完成以下目的：目标1。小头畸形的基本细胞机制是什么在患有IER3IP1 P.L78p突变的Meds患者中看到？ AIM2。IER3IP1是否调节分子对神经发育重要的膜受体的运输？该提议将填补我们的空白了解蛋白质运输如何调节神经发生并提供有关如何异常蛋白的洞察力贩运导致结构性大脑异常。