Modeling the neurovascular unit using Huntington's disease iPS cells

使用亨廷顿舞蹈病 iPS 细胞模拟神经血管单元

• 批准号: 8979452
• 负责人: Ryan Gar-Lok Lim
• 金额: $ 4.36万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-03-01 至 2017-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8979452
• 关键词: Affect; Animal Model; Apolipoprotein E; Astrocytes; Biodistribution; Blood - brain barrier anatomy; Blood Vessels; Brain; Bypass; CAG repeat; Cells; Cerebrovascular system; Chromosome Mapping; Coculture Techniques; Code; Cognitive; Computational Biology; Development; Disease; Disease model; Employee Strikes; Gene Expression; Gene Targeting; Genes; Genetic; Genetic Transcription; Homeostasis; Huntington Disease; Immunofluorescence Immunologic; In Situ Hybridization; Inherited; Maintenance; Messenger RNA; Modality; Modeling; Molecular; Movement; Neuraxis; Neurodegenerative Disorders; Neuronal Dysfunction; Neurons; Pathogenesis; Pathology; Pathway interactions; Patients; Play; Proteins; Research; Role; Signal Pathway; Signal Transduction; Stem cells; Structure; System; Testing; Tracer; base; brain endothelial cell; cell type; cerebral blood volume; disorder control; in vitro Model; in vivo; in vivo Model; induced pluripotent stem cell; innovation; middle age; mouse model; nervous system disorder; neuron loss; neurovascular unit; novel; public health relevance; response; stem cell technology

 DESCRIPTION (provided by applicant): Huntington's disease (HD) is a devastating, neurodegenerative disease that typically strikes in midlife and is caused by a CAG repeat expansion within the coding region of the HD gene. Due to the genetic basis of HD, unique opportunities exist to model the disease and extend scientific findings to other neurodegenerative diseases. Currently, there is no disease modifying treatment for any neurodegenerative disease, therefore it is critical to elucidate the cellular and molecular mechanisms leading to pathogenesis. Studies have primarily focused on studying the internal signals which prompt neuronal loss; however each cell in the neurovascular unit (NVU) plays a vital role in controlling homeostasis of the CNS and blood brain barrier (BBB) integrity. Utilizing induced pluripotent stem cell (iPSCs) technologies, one can generate in vitro models of HD neurons and other cells of the NVU, including brain endothelial cells (BECs) and astrocytes. Recent evidence of elevated arteriolar cerebral blood volume in HD patients, and increased vascularity in a mouse model of HD, suggests that the NVU and BBB may contribute to HD pathology. However understanding the specific genes involved and how they affect BBB functionality remain to be elucidated. It is known that Shh, Wnt, ApoE, and Ang-I/II signaling maintain homeostasis of the NVU; however, the specific contributions of these signaling pathways are not yet clear, especially in response to disease. It is important to note that both Shh and Wnt signaling, two pathways utilized by astrocytes and BECs in the NVU, are implicated in HD. We therefore hypothesize that abnormal signaling exists in and between BECs, astrocytes and neurons which causes abnormal expression of BBB proteins and contributes to neuronal dysfunction and pathogenesis in HD. Here we will examine if alterations in signaling and transcription exist between the cellular subunits of the NVU, and how this may affect BBB maintenance and subsequently induce neuronal damage in HD. We will utilize patient-derived iPSCs and computational biology approaches to create a model of the NVU in HD; focusing on the signaling between BEC and astrocytes. Specific Aims are to: Aim 1: Determine the contributions of gene networks and cell types in the NVU to disruption of the BBB using HD and control iPSCs. Aim 2: Examine the BBB in an in vivo murine model of HD.

 描述（由申请人提供）：亨廷顿氏病（HD）是一种毁灭性的神经退行性疾病，通常在中年发病，由HD基因编码区内的CAG重复扩增引起。由于HD的遗传基础，存在独特的机会来模拟疾病并将科学发现扩展到其他神经退行性疾病。目前，对于任何神经退行性疾病都没有疾病修饰治疗，因此阐明导致发病机制的细胞和分子机制至关重要。研究主要集中在研究促使神经元损失的内部信号;然而，神经血管单元（NVU）中的每个细胞在控制CNS和血脑屏障（BBB）完整性的稳态中起着至关重要的作用。利用 利用诱导多能干细胞（iPSC）技术，可以产生HD神经元和NVU的其他细胞（包括脑内皮细胞（BEC）和星形胶质细胞）的体外模型。最近的证据表明，HD患者的小动脉脑血容量升高，以及HD小鼠模型中的血管分布增加，表明NVU和BBB可能有助于HD病理学。然而，了解所涉及的特定基因以及它们如何影响BBB功能仍有待阐明。已知Shh、Wnt、ApoE和Ang-I/II信号传导维持NVU的稳态;然而，这些信号传导途径的具体贡献尚不清楚，特别是在响应疾病时。值得注意的是，Shh和Wnt信号传导，NVU中星形胶质细胞和BEC利用的两种途径，都与HD有关。因此，我们推测，异常信号存在于BEC，星形胶质细胞和神经元之间，导致BBB蛋白的异常表达，并有助于神经元功能障碍和发病机制在HD。在这里，我们将检查是否存在NVU的细胞亚基之间的信号转导和转录的改变，以及这可能如何影响血脑屏障的维持，并随后诱导HD中的神经元损伤。我们将利用患者来源的iPSC和计算生物学方法来创建HD中NVU的模型;重点关注BEC和星形胶质细胞之间的信号传导。具体目标是：目标1：使用HD和对照iPSC确定NVU中的基因网络和细胞类型对破坏BBB的贡献。目的2：在HD的体内小鼠模型中检查BBB。