Investigation of Pitt-Hopkins Syndrome pathophysiology using a human model

使用人体模型研究皮特霍普金斯综合症的病理生理学

基本信息

批准号：
10553718
负责人：
Alysson R. Muotri
金额：
$ 39.5万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN DIEGO
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-03-05 至 2025-12-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10553718
关键词：
Affect Anatomy Animal Model Architecture Back Biochemical Process Brain Cell Line Cell model Cells Cerebrum ChIP-seq Characteristics Child Childhood Clustered Regularly Interspaced Short Palindromic Repeats Communities Constipation Data Defect Development Disease Down-Regulation Drug Screening Electrophysiology (science)Epigenetic Process Exhibits Face Family Functional disorder Future Genes Genetic Genetic Diseases Genetic Predisposition to Disease Genome Goals Healthcare Systems Human Impairment In Vitro Individual Intellectual functioning disability Investigation Knowledge Lead Link Mediating Modeling Molecular Morphology Motor Mutate Mutation Nature Nervous System Neurologic Symptoms Neuronal Differentiation Neurons Organoids Outcome Parents Pathologic Pathway interactions Patients Phenotype Physiological Pitt-Hopkins syndrome Proliferating Property Rodent Speech Structure Symptoms TCF4 haploinsufficiency TCF7L2 gene Testing Tissues WNT Signaling Pathway autism spectrum disorder autistic behaviour autistic children cell type clinically relevant de novo mutation electrical property experimental study gastrointestinal gene therapy genetic approach human model in vitro Model induced pluripotent stem cell mouse model mutation correction nerve stem cell neural neurodevelopment neuropathology overexpression pharmacologic prevent progenitor repetitive behavior senescence severe intellectual disability single-cell RNA sequencing synaptogenesis therapeutic target transcription factor transcriptome sequencing

项目摘要

PROJECT SUMMARY Autism-spectrum disorders impact millions of individuals worldwide, representing a heavy toll on affected children, their families, and the health care system. Pitt–Hopkins Syndrome (PTHS) is an ASD caused by de novo mutations in the TCF4 gene. PTHS is characterized by severe intellectual disability, pronounced developmental and motor delays, absence of speech, repetitive behaviors, peculiar facial gestalt, and gastrointestinal manifestations. While the genetic etiology of PTHS is well established, the cellular and neural phenotypic alterations in human patients are still not fully understood, nor is it clear how TCF4 mutations cause such abnormalities. Lack of understanding about PTHS's molecular and cellular mechanisms is a problem because, until this information becomes available, specific altered pathways cannot be therapeutically targeted. Moreover, without neuropathological knowledge, it is impossible to treat and eventually cure PTHS by directly correcting the mutation in the genome. Our long-term goal is to understand how specific genetic defects and altered pathways in the brain result in the debilitating phenotypes exhibited by autistic children. The objectives of this application are to: (a) use human models of neural development in vitro to define the cellular and neural pathological consequences of clinically relevant TCF4 mutations in PTHS; and (b) provide proof-of-concept that correctional molecular strategies can be used to fix TCF4 expression, an approach that could eventually be used as gene therapy for PTHS. Our central hypothesis is that TCF4 mutations cause aberrant phenotypes in specific cell types of the nervous system, leading to the patients' neurological symptoms. We postulated that patient-derived in vitro models of PTHS can better recapitulate the pathophysiology than mouse models, because brain structure, genome architecture and development vary greatly between rodents and humans, and current PTHS animal models do not closely mimic all the disease's clinically relevant aspects. In preliminary experiments, we obtained patient-derived brain organoids and cultured neural cell types in vitro and used them as human models to show that PTHS neural progenitor cells exhibit senescence and decreased proliferation, accompanied by downregulation of Wnt signaling and SOX3 expression. Moreover, we observed that PTHS brain organoids fail to develop normal anatomically organized progenitor structures and that PTHS neurons display severely impaired firing properties. Our anticipated results/deliverables include the identification and manipulation of specific altered molecular pathways and neural cell types and the testing of genetic correctional strategies for the disease, which could propel future research on pharmacological and gene therapy for PTHS.

项目概要自闭症谱系障碍影响着全世界数以百万计的人，对受影响的人造成了沉重的损失儿童、他们的家庭和医疗保健系统皮特-霍普金斯综合症 (PTHS) 是一种由 de 引起的自闭症谱系障碍 (ASD)。 TCF4 基因的新突变的特点是严重的智力障碍。发育和运动迟缓、缺乏言语、重复行为、特殊的面部格式塔，以及虽然 PTHS 的遗传病因已明确，但细胞和神经系统的表现。人类患者的表型改变仍未完全了解，也不清楚 TCF4 突变是如何发生的缺乏对 PTHS 分子和细胞机制的了解是造成这种异常的原因之一。问题在于，在获得这些信息之前，无法对特定途径进行治疗而且，如果没有神经病理学知识，就不可能治疗并最终治愈 PTHS。通过直接纠正基因组中的突变。我们的长期目标是了解大脑中特定的遗传缺陷和通路是如何产生的在自闭症儿童所表现出的衰弱表型中，本应用的目的是：(a)使用。体外神经发育的人体模型，用于定义细胞和神经病理后果 PTHS 中临床相关的 TCF4 突变；以及 (b) 提供校正分子的概念验证策略可用于修复 TCF4 表达，这种方法最终可用作基因治疗 PTHS。我们的中心假设是 TCF4 突变会导致特定细胞类型的异常表型。神经系统，导致患者的神经系统症状我们推测源自患者体外。 PTHS 模型比小鼠模型能更好地概括病理生理学，因为大脑结构、啮齿动物和人类以及当前的 PTHS 动物之间的基因组结构和发育差异很大在初步实验中，模型并不能完全模拟该疾病的所有临床相关方面。获得了源自患者的大脑类器官，并在体外培养了神经细胞类型，并将它们用作人类模型显示 PTHS 神经祖细胞表现出衰老和增殖减少，此外，我们还观察到 PTHS 伴随着 Wnt 信号传导和 SOX3 表达的下调。大脑类器官无法发育出正常的解剖学组织的祖细胞结构，并且 PTHS 神经元显示严重受损的烧制性能。我们的预期结果/交付成果包括识别和特定分子途径和神经细胞类型的操纵以及遗传测试该疾病的纠正策略，这可以推动未来的药理学和遗传学研究 PTHS 的治疗。