Characterization and Early Assays Development in FOXG1 Deficient Neurons

FOXG1 缺陷神经元的表征和早期检测开发

• 批准号: 10727503
• 负责人: JED L HUBBS
• 金额: $ 17.7万
• 依托单位: BOSTON CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10727503
• 关键词: Biological Assay; Boston; CRISPR/Cas technology; Cell Differentiation process; Cell Line; Cells; Clinical; Development; Developmental Delay Disorders; Disease; Dyes; Ethics; Fluorescent Dyes; Future; Gene Proteins; Genes; Genetic; Genotype; Goals; Heterozygote; Image; Individual; Interneurons; Ligands; Measures; Methods; Microcephaly; Movement; Muscle hypotonia; Mutation; Neurodevelopmental Disorder; Neurologic Symptoms; Neurons; Parents; Pathway interactions; Patients; Pediatric Hospitals; Pharmaceutical Preparations; Pharmacotherapy; Pluripotent Stem Cells; Protein Deficiency; Proteins; Reporter; Reporter Genes; Reporting; Research; Seizures; Somatic Cell; Stereotyping; Symptoms; Syndrome; System; Telencephalon; Validation; Work; assay development; drug discovery; gene therapy; genomic locus; induced pluripotent stem cell; interest; mutant; nerve stem cell; nervous system disorder; postnatal development; pre-clinical; programs; protein expression; screening; severe intellectual disability; small molecule; small molecule therapeutics; stem cell proliferation; symptomatic improvement; therapeutic development; therapy development; transcription factor

PROJECT SUMMARY FOXG1 syndrome is a rare neurological disorder that often causes severe intellectual disability, microcephaly and neurological symptoms including hypotonia, seizures, and stereotypic movements. FOXG1 syndrome is caused by heterozygous mutation or deletion of the forkhead box 1 protein (FOXG1), a transcription factor that controls expression of key cortical development proteins. FOXG1 is specifically important for telencephalon neural progenitor cell (NPC) proliferation and differentiation to GABAergic interneurons. We have prioritized FOXG1 syndrome based on a framework we developed to evaluate monogenetic neurodevelopmental disorders for therapeutic development based on generic, preclinical validation, clinical and ethical considerations. We have also created three pluripotent stem cell (PSC) lines from FOXG1 syndrome patients and used CRISPR/Cas9 gene editing to create isogenic control lines. We hypothesize that a drug-like small molecule that increases FOXG1 expression in patients during early postnatal development would improve symptoms in these individuals. In the proposed research, we will use FOXG1 PSC lines to create an endogenous locus gene reporter line by using CRISPR/Cas9 to insert a HaloTag reporter protein at the C-terminus of the FOXG1 gene. A method will be developed whereby these modified FOXG1 PSCs with be differentiated to cortical neural progenitor cells (NPCs) and treated with HaloTag ligand tethered dyes to measure accurately measure FOXG1 expression. The primary goal of this project will be to demonstrate that this FOXG1 reporter system is suitable for screening small molecules for their ability to increase FOXG1 expression.

项目概要 FOXG1 综合征是一种罕见的神经系统疾病，通常会导致严重的智力障碍， 小头畸形和神经系统症状，包括肌张力减退、癫痫发作和刻板印象 动作。 FOXG1综合征是由叉头杂合突变或缺失引起的 box 1 蛋白 (FOXG1)，一种控制关键皮质发育表达的转录因子 蛋白质。 FOXG1 对于端脑神经祖细胞 (NPC) 特别重要 增殖和分化为 GABA 能中间神经元。我们优先考虑 FOXG1 综合征 基于我们开发的评估单基因神经发育障碍的框架 基于仿制药、临床前验证、临床和伦理的治疗开发 考虑因素。我们还从 FOXG1 中创建了三个多能干细胞 (PSC) 系 综合征患者并使用 CRISPR/Cas9 基因编辑创建等基因对照系。我们 假设一种药物样小分子可以增加患者体内 FOXG1 的表达 产后早期发育将改善这些人的症状。在提议的 研究中，我们将使用 FOXG1 PSC 系来创建内源基因座基因报告系 使用 CRISPR/Cas9 在 FOXG1 基因的 C 末端插入 HaloTag 报告蛋白。 将开发一种方法，使这些修饰的 FOXG1 PSC 能够分化为 皮层神经祖细胞 (NPC) 并用 HaloTag 配体系留染料处理以测量 准确测量 FOXG1 表达。该项目的主要目标是展示 该 FOXG1 报告系统适合筛选小分子的能力 增加 FOXG1 表达。