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: 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
• 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

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.

项目总结 自闭症谱系障碍影响着全球数百万人，这对受影响的人来说是沉重的代价。 儿童、他们的家庭和医疗保健系统。皮特-霍普金斯综合征(PTHS)是由阿尔茨海默病引起的自闭症。 TCF4基因的新突变。PTHS的特点是严重的智力残疾，明显的 发育和运动迟缓，不说话，重复行为，特殊的面部格式塔，以及 胃肠道症状。虽然PTHS的遗传病因已经确立，但细胞和神经 人类患者的表型改变仍然不完全清楚，也不清楚TCF4突变是如何 造成这种反常现象。对PTHS的分子和细胞机制缺乏了解是一种 问题是，在获得这些信息之前，特定的改变的通路不能用于治疗 有针对性的。此外，如果没有神经病理学知识，治疗并最终治愈PTHS是不可能的。 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基因缺陷和改变的路径是如何导致的 在自闭症儿童表现出的衰弱表型中。本申请的目的是：(A)使用 人类神经发育的体外模型，以确定细胞和神经病理后果 临床相关的PTHS中的TCF4突变；以及(B)提供纠正分子的概念证明 可以使用策略来修复TCF4的表达，这种方法最终可能被用于基因治疗 PTHS。我们的中心假设是TCF4突变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。我们推测患者在体外衍生的 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目前的PTHS动物之间的基因组结构和发育差异很大 模型并不能很好地模拟这种疾病的所有临床相关方面。在初步实验中，我们 获得患者来源的脑组织和体外培养的神经细胞类型，并将其用作人类 显示PTHS神经前体细胞衰老和增殖减少的模型， 伴有Wnt信号和SOX3表达下调。此外，我们观察到PTHS 脑器官不能发育正常的解剖学组织的前体结构，PTHS神经元 显示严重受损的射击性能。我们的预期成果/交付成果包括确定和 特定改变的分子通路和神经细胞类型的操纵以及基因检测 疾病的纠正策略，这可能会推动未来的药理和基因研究 PTHS的治疗。