Identification of TSC cellular phenotypes using patient-derived iPSCs

使用患者来源的 iPSC 鉴定 TSC 细胞表型

• 批准号: 8790825
• 负责人: GABRIELLA D'ARCANGELO
• 金额: $ 22.93万
• 依托单位: RUTGERS, THE STATE UNIV OF N.J.
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-30 至 2016-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8790825
• 关键词: Affect; Autistic Disorder; Biochemical; Biological Models; Blood specimen; Brain Diseases; Cell Line; Cell Nucleus; Cell model; Cell physiology; Cells; Complex; Cortical Malformation; Dendrites; Development; Disease; Epilepsy; Etiology; Exhibits; Frequencies; Genes; Genetic; Genome; Growth Cones; Hand; Image; Image Analysis; Impaired cognition; Individual; Intellectual functioning disability; Lead; Microscopy; Modeling; Morphology; Mutation; Neurologic; Neurologic Manifestations; Neurons; Organ; Patients; Peripheral Blood Lymphocyte; Phenotype; Phosphorylation; Phosphotransferases; Predisposition; Process; Proteins; Rare Diseases; Regulation; Rodent; Severities; Siblings; Signal Transduction; Structure; Symptoms; Synapses; Synaptic plasticity; Syndrome; System; TSC1 gene; TSC2 gene; Techniques; Technology; Tuberous sclerosis protein complex; Work; autistic behaviour; axon growth; brain cell; brain malformation; density; developmental disease; effective therapy; human FRAP1 protein; in vitro Model; induced pluripotent stem cell; molecular phenotype; neuronal cell body; novel; public health relevance; synaptogenesis; trait; tumor

 DESCRIPTION (provided by applicant): Tuberous sclerosis complex (TSC) is a developmental disorder characterized by tumor susceptibility in multiple organs, brain malformations, and neurological manifestations. Despite considerable progress in understanding the genetic and signaling mechanisms underlying the disease, effective treatments are still lacking, particularly with regard to the control of neurological symptoms. In this proposal, we plan to develop induced pluripotent stem cell (iPSC) lines from patient and unaffected siblings, and differentiate these cells to generate neuronal cultures as a novel in vitr system to identify cellular and molecular phenotypes of the disease. Since the patients carry identified, de novo, heterozygous mutations in the TSC2 gene, we will also attempt to correct the genetic TSC mutations in iPSCs using the TALEN technology. With these iPSC models at hand, we will determine whether heterozygous TSC2 neurons exhibit a morphological or synaptic phenotype. Second, we will determine whether they exhibit localized alterations in signal transduction complexes that are normally regulated by the TSC, such as mTORC1 and mTORC2. We hypothesize that heterozygous TSC2 neurons express a subtle morphological phenotype that results from the localized de-regulation of mTOR-containing signaling complexes. This phenotype may be key to the etiology of cognitive dysfunction and autism in TSC patients. TSC is a relatively rare disorder affecting approximately 1 in 6,000 individuals. However, it shares mechanistic underpinnings with a number of cortical malformation syndromes, and it is frequently associated with epilepsy (>90%), intellectual disability and autism (40-50%). Thus, our findings are relevant to the treatment of all these developmental brain disorders, extending the potential impact of our work beyond the field of TSC.

 描述（申请人提供）：结节性硬化症（TSC）是一种发育障碍，其特征是多个器官的肿瘤易感性、脑畸形和神经系统表现。尽管在了解该疾病的遗传和信号传导机制方面取得了相当大的进展，但仍然缺乏有效的治疗方法，特别是在控制神经系统症状方面。在这项提案中，我们计划从患者和未受影响的兄弟姐妹中开发诱导多能干细胞（iPSC）系，并分化这些细胞以产生神经元培养物，作为一种新型的玻璃体系统，以识别该疾病的细胞和分子表型。由于患者携带已识别的 TSC2 基因从头杂合突变，我们还将尝试使用 TALEN 技术纠正 iPSC 中的 TSC 基因突变。有了这些 iPSC 模型，我们将确定杂合的 TSC2 神经元是否表现出形态或突触表型。其次，我们将确定它们是否表现出通常由 TSC 调节的信号转导复合物（例如 mTORC1 和 mTORC2）的局部改变。我们假设杂合的 TSC2 神经元表达一种微妙的形态表型，这是由含有 mTOR 的信号复合物的局部失调引起的。这种表型可能是 TSC 患者认知功能障碍和自闭症病因的关键。 TSC 是一种相对罕见的疾病，大约每 6,000 人中就有 1 人受到影响。然而，它与许多皮质畸形综合征具有共同的机制基础，并且经常与癫痫（>90%）、智力障碍和自闭症（40-50%）相关。因此，我们的研究结果与所有这些发育性脑部疾病的治疗相关，将我们工作的潜在影响扩展到 TSC 领域之外。