Epigenetics-Based Autism Treatment with Animal Models and Human Stem Cells

利用动物模型和人类干细胞进行基于表观遗传学的自闭症治疗

• 批准号: 10651463
• 负责人: JIAN FENG
• 金额: $ 61.57万
• 依托单位: STATE UNIVERSITY OF NEW YORK AT BUFFALO
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-02-15 至 2028-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10651463
• 关键词: ASD patient; Address; Animal Model; Autopsy; Behavioral; Biochemical; ChIP-seq; Chromatin; Corpus striatum structure; DNA Sequence Alteration; Defect; Electrophysiology (science); Enzymes; Epigenetic Process; Etiology; Exhibits; Exons; Fibroblasts; Functional disorder; Gene Activation; Gene Expression; Gene Expression Alteration; Genes; Genetic; Genetic Transcription; Genetic study; Genomics; Glutamates; Goals; Heterozygote; Histones; Human; Human Genetics; Impairment; Induced pluripotent stem cell derived neurons; Intervention; KDM1A gene; Large-Scale Sequencing; Length; Link; Lysine; Mediating; Methylation; Modeling; Molecular; Molecular Abnormality; Mus; Neurodevelopmental Disorder; Neuronal Differentiation; Neurons; Pathogenicity; Patients; Phelan-McDermid syndrome; Phenotype; Play; Prefrontal Cortex; Proteins; Research; Risk Factors; Role; Scaffolding Protein; Social Interaction; Symptoms; Synapses; Testing; Therapeutic; Therapeutic Effect; Tissues; Transcriptional Regulation; Translating; Work; autism spectrum disorder; autistic; demethylation; drug discovery; gene repression; genome-wide; high risk; histone demethylase; histone methylation; histone methyltransferase; histone modification; human stem cells; induced pluripotent stem cell; inhibitor; innovation; interdisciplinary approach; knock-down; loss of function mutation; mouse model; neuronal excitability; novel; novel therapeutic intervention; permissiveness; pharmacologic; repetitive behavior; response; risk variant; side effect; social deficits; stem cell differentiation; stem cell technology; stem cells; synaptic function; targeted agent; targeted treatment; transcription factor; transcriptome sequencing; treatment strategy

Summary This project aims to discover novel pharmacological intervention for core symptoms of autism, including social deficits and repetitive behaviors. One of the causal factors of autism is the loss of Shank3 gene, which encodes a scaffolding protein at glutamatergic synapses. We will use Shank3-deficient mouse models and human stem cell-derived neurons in this drug discovery endeavor. Genetics studies have found that many of genes disrupted in autism are histone-modifying enzymes that mediate histone methylation/demethylation, which play a key role in transcriptional regulation. Our preliminary studies have found that histone lysine 4 dimethylation (H3K4me2, linked to gene activation) is significantly decreased in the prefrontal cortex (PFC) of autistic humans and Shank3-deficient mice. H3K4me2 is demethylated by lysine-specific histone demethylase 1 (LSD1, KDM1A), which is found to be increased in PFC neurons of Shank3-deficient mice. We hypothesize that inhibiting LSD1 to elevate H3K4me2 and restore gene expression may be able to ameliorate autism-like phenotypes, therefore providing a novel therapeutic strategy for autism. Combined behavioral, biochemical, electrophysiological, genomic and stem cell approaches will be used to test this hypothesis. Aim 1, we will characterize epigenetic changes and therapeutic effects of epigenetic agents in mouse models of autism. The alteration of histone methylation marks and histone demethylases will also be examined in PFC of Shank3- deficient mice and autism human postmortem tissues. Aim 2, we will reveal the molecular mechanisms underlying epigenetic treatment of autism models. Synaptic responses and neuronal excitability will be recorded in Shank3-deficient mice treated with LSD1 inhibitors. Genome-wide alteration of gene expression and histone methylation will be examined using RNAseq and ChIPseq. The causal role of identified key molecules in the therapeutic effects of LSD1 inhibitors will also be determined. In Aim 3, we will examine the molecular alteration and treatment strategy in human neurons from ASD patient with Shank3 haploinsufficiency. To find out whether the epigenetic treatment strategy found in Shank3 mouse models might also work in autism patients, we will use the innovative stem-cell technology to examine the capability of LSD1 inhibitors to reverse synaptic deficits and molecular aberrations in ASD patient’s neurons derived from induced pluripotent stem cells. Results from this study will not only reveal the mechanistic link among important autism risk factors, but also uncover a mechanism-based treatment strategy for autism.

概括 该项目旨在发现自闭症核心症状的新型药理干预，包括社会 缺陷和重复行为。自闭症的因果因素之一是尚克3基因的丧失， 在谷氨酸能突触中编码脚手架蛋白。我们将使用shank3缺陷鼠标模型，并且 在这项药物发现努力中，人类干细胞衍生的神经元。遗传学研究发现 自闭症中破坏的基因是组蛋白修饰的酶，可介导组蛋白甲基化/脱甲基化的酶， 在转录调节中起关键作用。我们的初步研究发现组蛋白赖氨酸4 二甲基化（H3K4ME2，与基因激活相关的H3K4ME2）在前额叶皮层（PFC）中显着降低 自闭症人和尚克3缺陷小鼠。 H3K4ME2通过赖氨酸特异性组蛋白脱甲基酶脱甲基化 1（LSD1，KDM1A），发现在Shank3缺陷小鼠的PFC神经元中增加了。我们假设 抑制LSD1升高H3K4ME2并恢复基因表达可能能够改善自闭症样 因此，表型为自闭症提供了一种新颖的治疗策略。联合行为，生化， 电生理，基因组和干细胞方法将用于检验该假设。目标1，我们将 表征表观遗传剂在自闭症小鼠模型中的表观遗传变化和治疗作用。这 在shank3-的PFC中，还将检查组蛋白甲基化标记和组蛋白脱甲基酶的改变 缺乏小鼠和自闭症人类验尸组织。 AIM 2，我们将揭示分子机制 自闭症模型的基础表观遗传治疗。突触反应和神经元会令人兴奋 记录在用LSD1抑制剂治疗的Shank3缺陷小鼠中。基因表达的全基因组改变 将使用RNASEQ和CHIPSEQ检查组蛋白甲基化。确定密钥的因果作用 还将确定LSD1抑制剂治疗作用中的分子。在AIM 3中，我们将检查 ASD患者Shank3的人类神经元中的分子改变和治疗策略 单倍不足。找出Shank3小鼠模型中发现的表观遗传治疗策略是否可能 同样在自闭症患者中工作，我们将使用创新的干细胞技术来检查LSD1的能力 反向诱导的ASD患者神经元中合成缺陷和分子像差的抑制剂 多能干细胞。这项研究的结果不仅会揭示重要的自闭症之间的机理联系 风险因素，但也发现了基于机制的自闭症治疗策略。