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：
10366017
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/10366017
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 health status 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 structure Neurologic Symptoms Neurons Organoids Outcome Parents Pathologic Pathway interactions Patients Pharmacology Pharmacotherapy Phenotype Physiological Pitt-Hopkins syndrome Property Rodent Speech Structure Symptoms TCF7L2 gene Testing Tissues WNT Signaling Pathway autism spectrum disorder autistic behaviour autistic children base 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 nerve stem cell neural model neurodevelopment overexpression prevent progenitor relating to nervous system 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.

项目摘要自闭症谱系障碍会影响全球数以百万计的人，压抑了受影响的巨大损失儿童，他们的家人和医疗保健系统。 TCF4基因中的Novo突变是严重的智力障碍的发育和运动的延迟，缺乏言语，重复行为，特殊的面部野蛮人以及虽然PTH的遗传病因已经很好地确定，但细胞和神经人类患者的表型改变仍未完全了解由于这种异常缺乏对PTH的分子和细胞机制的理解问题是因为，在这些信息可用之前，特定的更改途径不能在治疗上进行此外，没有神经病理学知识，无法治疗通过直接纠正基因组中的突变。我们的长期目标是了解特定的遗传缺陷和大脑结果中的途径如何改变在自闭症儿童中表现出的衰弱的表型中。人体在体外神经发育的模型，以定义细胞和神经病理的结构 PTH和（b）在临床上相关的TCF4突变提供了校正分子的证明策略可用于修复TCF4表达，这种方法最终可以用作基因疗法的ASED ASED ASED PTH。我们的中心假设是TCF4突变在特定细胞类型中引起异常表型神经系统，导致患者的神经内在症状。与小鼠模型相比啮齿动物和人类之间的基因组结构和发展差异很大，当前的PTHS动物模型在初步实验中并没有密切模仿该疾病的所有临床方面。在体外获得了患者衍生的脑器官和培养的神经细胞类型，并将其用作人类 PTHS神经祖细胞的模型表现出衰老和增殖降低，伴随着Wnt信号的下调和SOX3表达。脑器官无法发展出正常的解剖结构，并且PTHS神经元显示严重损害的发射属性。操纵特定改变的分子途径和神经细胞细胞类型以及遗传测试该疾病的矫正策略，可以推动对药理学和基因的未来研究 PTH的治疗。