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

• 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
• 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

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百万个人，代表受影响的巨大损失 儿童，他们的家人和医疗保健系统。皮特 - 霍金斯综合征（PTHS）是由DE引起的ASD TCF4基因中的Novo突变。 PTH的特征是严重的智力残疾，发音为 发育和运动的延迟，缺乏言语，重复行为，特殊的面部护理和 胃肠道表现。虽然PTH的遗传病因已良好，但细胞和中性 人类患者的表型改变仍未完全了解，也不清楚TCF4突变 导致这种异常。对PTH的分子和细胞机制缺乏了解是一种 问题是因为，在此信息可用之前，特定的更改途径不能热吗 目标。此外，没有神经病理学知识，就无法治疗，有时无法治愈PTH 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 在加速儿童暴露的衰弱的表型中。此应用程序的目标是：（a）使用 在体外神经元发育的人类模型，以定义细胞和神经病理学的后果 PTH中与临床相关的TCF4突变； （b）提供矫正分子的概念验证 策略可用于修复TCF4表达，这种方法最终可以用作基因治疗 PTHS。我们的中心假设是TCF4突变在特定细胞类型的特定细胞类型中引起异常表型 神经系统，导致患者的神经系统症状。我们假设患者衍生的体外 与小鼠模型相比，PTH的模型可以更好地概括病理生理学，因为大脑结构， 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 模型并未密切模仿该疾病与临床相关的所有方面。在初步实验中，我们 在体外获得了患者衍生的脑器官和培养的神经元细胞类型，并将其用作人类 模型表明PTHS神经祖细胞暴露了感应并改善增殖， 伴随着Wnt信号传导和SOX3表达的下调。而且，我们观察到PTH 脑器官无法发展出正常的解剖结构，并且PTHS神经元 显示严重受损的射击属性。我们预期的结果/可交付成果包括标识和 操纵特定改变的分子途径和神经细胞类型以及通用测试 该疾病的矫正策略，可以推动对药物和基因的未来研究 PTH的治疗。