Epigenetic mechanisms of disrupted neurodevelopment in Menke-Hennekam syndrome

Menke-Hennekam 综合征神经发育障碍的表观遗传机制

• 批准号: 10816703
• 负责人: Artem Barski
• 金额: $ 16.05万
• 依托单位: CINCINNATI CHILDRENS HOSP MED CTR
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-15 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10816703
• 关键词: Acceleration; Acetylation; Address; Astrocytes; Automobile Driving; Behavior; Bioinformatics; Biological Assay; Brain; Brain Diseases; CREBBP gene; Calcium Signaling; Cell Line; ChIP-seq; Characteristics; Chromatin Remodeling Factor; Collaborations; Data; Defect; Dendrites; Development; Diagnosis; Disease; Disease model; EP300 gene; Ectoderm; Enhancers; Epigenetic Process; Exons; FDA approved; Face; Foundations; Frequencies; Future; Gene Expression; Genes; Glutamates; Hennekam syndrome; Heterochromatin; Heterozygote; Histone Acetylation; Human; Individual; Intellectual functioning disability; Knowledge; Left; Length; Measures; Mediating; Methods; Microcephaly; Microglia; Missense Mutation; Modeling; Molecular; Molecular Target; Mutate; Mutation; Neocortex; Nerve Degeneration; Nervous System; Neuroglia; Neurologic; Neuronal Differentiation; Neurons; Organ; Pathway interactions; Patients; Pharmaceutical Preparations; Phenotype; Pilot Projects; Play; Population; Prefrontal Cortex; Proteins; RNA; Radial; Reporting; Research; Role; Seizures; Signal Transduction; Solid; Specificity; Symptoms; Synapses; Technical Expertise; Testing; Time; Transcription Coactivator; Variant; Vertebral column; Work; autism spectrum disorder; cell type; comparison control; craniofacial; directed differentiation; druggable target; epigenetic regulation; epigenomics; excitatory neuron; gene network; histone acetyltransferase; human pluripotent stem cell; induced pluripotent stem cell; interest; malformation in cortical development; neocortical; nerve stem cell; nervous system disorder; neurodevelopment; neuroinflammation; neuron loss; paralogous gene; postsynaptic; presynaptic; promoter; public health relevance; repetitive behavior; stem; stem cell differentiation; synaptogenesis

Project Summary Heterozygous variants in EP300, a histone acetyltransferase, cause two rare multi-organ chromatinopathies: Menke-Hennekam syndromes type 2 (MKHK2; MIM618333, specifically mutations in exon 30-31) and Rubinstein-Taybi type 2 (RSTS2; MIM613684). Interestingly, based on “inverse” patient craniofacial characteristics with RSTS and the alignment of MKHK2 facial features with another disorder with duplicated regions of CREBBP (EP300's paralog), authors have proposed mutations causing MKHK and RSTS have potentially different functions (i.e., gain [GOF] versus loss [LOF], respectively).1,2 In addition, individuals with MKHK2 have overlapping manifestations with disorders with malformations in cortical development (MCD), such as microencephaly, seizures, intellectual disability, repetitive behaviors, and increased rates of co-diagnoses with autism spectrum disorder, demonstrating a potential role in cortical neuron function.3 While the specialized exon 30-31 missense mutations found in MKHK2 have yet to be modeled, RSTS2 LOF models have yielded phenotypic changes of shorter branches and hypo-excitability. However, the RSTS2 LOF bulk neuronal bioinformatical analyses contained contaminating cell types with differential regional specificity, therefore, EP300 specific gene network/pathways are uninterpretable. Together this demonstrates a critical need to understand EP300's role in maturation and function of cortical excitatory neurons before treatment of MKHK2 neurological issues can be tackled. We hypothesize that MKHK2 EP300 mutation causes accelerated maturation rate, decreased dendrite formation, and altered neuronal function. This proposal is a collaboration between Drs. Potter and Barski who specialize in epigenomics and Dr. Tchieu, a developmental biologist who developed methods for human induced pluripotent stem cell (hPSC) differentiation into nervous system cell types. In this study, we will use this approach to differentiate MKHK2 mutated EP300 and corresponding isogenic control hPSCs into radial glia-like neural stem cells and PFC excitatory neurons in order to: (1) identify MKHK2-related defects in neuronal maturation and signaling and (2) to identify epigenetic and gene expression changes driving them. This pilot project will provide a solid foundation for the study of the mechanism(s) behind neurocortical developmental defects in MKHK2 and allow identification of molecular targets for potential therapies for MKHK2 seizures and other related MCD-like symptoms. Further, we can apply knowledge and technical expertise gained from this project to other epigenetic proteins to further expand our knowledge of epigenetic regulation during corticogenesis, specifically neuronal maturation and signaling allowing for a broader impact on the treatment of MCD, neurodegenerative, neurodevelopment, and neuroinflammatory disorders.

项目摘要 EP300（一种组蛋白乙酰转移酶）中的杂合变体引起两种罕见的多器官染色质病： Menke-Hennekam综合征2型（MKHK2; MIM618333，特别是外显子30-31中的突变）和 Rubinstein-Taybi 2型（RSTS2; MIM613684）。有趣的是，基于“逆”患者颅面 rsts的特征以及MKHK2面部特征的对齐，并具有另一种重复的疾病 CREBBP（EP300的Paralog）区域，作者提出了导致MKHK和RST的突变 潜在的不同功能（即获得[GOF]和损失[LOF]）。1,2此外，患有 MKHK2具有与皮质发育中畸形（MCD）的疾病重叠的表现， 例如微脑，癫痫发作，智力残疾，重复行为和共同诊断率提高 患有自闭症谱系障碍，表现出在皮质神经元功能中的潜在作用。3虽然专业 在MKHK2中发现的外显子30-31错义突变尚未建模，RSTS2 LOF模型已产生 较短的分支和缺点的表型变化。但是，RSTS2 LOF散装神经元 生物信息学分析包含具有不同区域特异性的污染细胞类型，因此 EP300特定的基因网络/途径是无法解释的。这共同表明了至关重要的需要 在治疗MKHK2之前，了解EP300在皮质兴奋神经元的成熟和功能中的作用 可以解决神经系统问题。我们假设MKHK2 EP300突变导致加速 成熟率，树突形成降低和神经元功能改变。 该建议是Drs之间的合作。专门从事表观基因组学的波特和巴斯基（Barski） 开发了人类诱导多能干细胞（HPSC）分化方法的发展生物学家 进入神经系统细胞类型。在这项研究中，我们将使用这种方法来区分MKHK2突变EP300 以及相应的异源对照HPSC中的radial胶质神经元细胞和PFC兴奋性神经元 命令：（1）识别神经元成熟和信号传导中MKHK2相关的缺陷以及（2）识别表观遗传 基因表达会改变驱动它们。 该试点项目将为研究神经皮质背后的机制提供坚实的基础 MKHK2中的发育缺陷，并允许鉴定潜在疗法的分子靶标的 MKHK2癫痫发作和其他相关的类似MCD的符号。此外，我们可以运用知识和技术 从这个项目获得的专业知识到其他表观遗传蛋白，以进一步扩大我们对表观遗传的了解 皮质生成期间的调节，特别是神经元成熟和信号传导，允许更广泛的影响 关于MCD的治疗，神经退行性，神经发育和神经炎性疾病。