Tooth pulp as a source for neuronal precursor cells to study neurogenetic disorde

牙髓作为神经元前体细胞的来源来研究神经遗传疾病

• 批准号: 8299874
• 负责人: LAWRENCE T REITER
• 金额: $ 18.73万
• 依托单位: UNIVERSITY OF TENNESSEE HEALTH SCI CTR
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-04-01 至 2014-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8299874
• 关键词: 15q; Alleles; Angelman Syndrome; Animal Model; Autistic Disorder; Autopsy; Biological Models; Biopsy; Brain; Cell Line; Cells; Cephalic; Characteristics; Cicatrix; DNA; Dental Pulp; Dentistry; Disabled Children; Disease; Distress; Epigenetic Process; Fibroblasts; Fragile X Syndrome; Future; Gene Expression; Gene Expression Regulation; Gene Proteins; Genes; Genetic; Goals; Hereditary Disease; Human; Human Genetics; In Vitro; Ion Channel; Lead; Measures; Methods; Methylation; Modeling; Molecular; Mus; Neural Crest; Neural Crest Cell; Neurons; Pain; Pathogenesis; Patients; Pattern; Peripheral Nervous System; Physiological; Physiology; Property; Sampling; Skin; Source; Stem cells; Sum; Synapses; Syndrome; Testing; Therapeutic; Tissues; Tooth structure; Transcript; Transplantation; Viral Vector; autism spectrum disorder; autistic children; base; brain tissue; cell type; deciduous tooth; developmentally disabled; immunogenic; imprint; in vivo; induced pluripotent stem cell; insight; lymphoblast; mouse model; neurogenetics; neuron development; neurophysiology; novel strategies; precursor cell; protein expression; receptor; research study; stem

DESCRIPTION (provided by applicant): The study of neurogenetic disorders like Angelman, Rett and fragile X syndromes as well as autism spectrum disorders has depended largely on the analysis of gene/protein expression in non-neuronal biospecimens like lymphoblast and fibroblast cell lines. Although some analysis has been possible in post-mortem human brain tissue for neurogenetic syndromes, this tissue is often of variable quality and in limited amounts. Recently there has been an initiative to induce pluripotent stem cells (iPSCs) from patient fibroblasts and differentiate these iPSCs into various neuronal lineages. There are several problems with this approach: 1) fibroblasts must be obtained though a fairly invasive skin biopsy which leaves a scar and causes undue pain and distress in developmentally disabled or autistic children; 3) induction of fibroblasts into stem cells (reprogramming) and then into neuronal lineages is a laborious task that may not maintain epigenetic marks on the DNA that are essential to proper gene regulation in the native neuronal tissues; and 4) viral vectors which are themselves immunogenic are used to reprogram iPSCs limiting the downstream aplication of these neurons for therapeutic transplantations. Here we propose to use dental pulps from exfoliated primary teeth (EPT), as a source of stem cells for the study of a variety of neurogenetic syndromes. Dental pulp stem cells differentiate into functional neurons in vitro and in vivo. Dental pulps from EPT are also an easily obtainable source of cranial neural crest (CNC) cells. The majority of the cells in the peripheral nervous system, including neurons, are derived from the neural crest. Based on the extensive collaborative sum of our expertise in genetics, dental pulp stem cells and neurophysiology, we feel that this novel approach will provide a fresh perspective to the way we study expresion changes, epigenetics and neurophysiology in an ex-vivo model system for human neurogenetic disorders. ! PUBLIC HEALTH RELEVANCE: The primary goal of this proposal is to develop a method to investigate the neurons of patients with neurogenetic disease that uses shed teeth (primary teeth or puled teeth). If we can determine that tooth samples provide the right type of cells in th dental pulp to make neurons in culture we will have a new way to look at the gene expression and physiology of these patient-derived neurons. Having these cultures could lead to new insights into the mechanisms of human neurogenetic disease and even normal neuronal development and function.

描述（由申请人提供）：对 Angelman、Rett 和脆性 X 综合征等神经遗传性疾病以及自闭症谱系障碍的研究很大程度上依赖于对非神经元生物样本（如淋巴母细胞和成纤维细胞系）中基因/蛋白质表达的分析。尽管可以对死后的人类脑组织进行神经遗传综合征的一些分析，但这种组织的质量往往参差不齐，而且数量有限。 最近，有一项计划从患者成纤维细胞中诱导多能干细胞 (iPSC)，并将这些 iPSC 分化为各种神经元谱系。这种方法存在几个问题：1）必须通过相当侵入性的皮肤活检来获取成纤维细胞，这会留下疤痕并导致发育障碍或自闭症儿童过度疼痛和困扰； 3) 将成纤维细胞诱导成干细胞（重编程），然后诱导成神经元谱系是一项艰巨的任务，可能无法维持 DNA 上的表观遗传标记，而这些标记对于天然神经元组织中的正确基因调控至关重要； 4) 本身具有免疫原性的病毒载体用于重新编程 iPSC，限制这些神经元用于治疗性移植的下游应用。在这里，我们建议使用脱落乳牙（EPT）的牙髓作为干细胞来源，用于研究各种神经遗传综合征。牙髓干细胞在体外和体内分化为功能性神经元。 EPT 牙髓也是颅神经嵴 (CNC) 细胞的容易获得的来源。周围神经系统中的大多数细胞，包括神经元，都源自神经嵴。基于我们在遗传学、牙髓干细胞和神经生理学方面的专业知识的广泛合作总结，我们认为这种新颖的方法将为我们在人类神经遗传疾病离体模型系统中研究表达变化、表观遗传学和神经生理学的方式提供一个新的视角。 ！ 公共健康相关性：该提案的主要目标是开发一种方法来研究使用脱落牙齿（乳牙或拔牙）的神经遗传疾病患者的神经元。如果我们能够确定牙齿样本在牙髓中提供了正确类型的细胞来在培养物中产生神经元，我们将有一种新的方法来观察这些源自患者的神经元的基因表达和生理学。拥有这些培养物可以带来对人类神经遗传疾病甚至正常神经元发育和功能机制的新见解。