Quantifying Cortical Neuron Production After Transplantation

移植后量化皮质神经元的产生

• 批准号: 7388436
• 负责人: Lawrence D Recht
• 金额: $ 21.52万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-15 至 2009-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7388436
• 关键词: Address; Age; Apical; Area; Axon; Behavior; Birth trauma; Brain; Brain Injuries; Cell Differentiation process; Cell Line; Cell Transplants; Cells; Cerebral Palsy; Cerebral cortex; Cervical; Characteristics; Chromosome Pairing; Clinic; Coculture Techniques; Collaborations; Complex; Computer information processing; Condition; Dendrites; Depth; Development; ES Cell Line; Effectiveness; Engineering; Event; Flow Cytometry; Gene Expression; Gene Silencing; Generations; Genes; Goals; Grant; Hypoxia; In Situ; In Vitro; Injury; Medical; Methods; Mus; Neonatal; Neonatal Brain Injury; Neuronal Differentiation; Neurons; Nuclear Pore Complex; Numbers; Output; Phenotype; Population; Positioning Attribute; Premature Infant; Process; Production; Quality of life; Recovery; Recovery of Function; Sensory; Sensory Process; Spinal Cord; Stem cell transplant; Stem cells; Stromal Cells; Structure; Synapses; Testing; Therapeutic; Transplantation; Trauma; Tretinoin; base; cell type; college; embryonic stem cell; functional improvement; hippocampal pyramidal neuron; implantation; improved; in vivo; interest; mouse model; neural circuit; pluripotency; precursor cell; promoter; reconstitution; relating to nervous system; repaired; research study; transcription factor; transmission process; white matter

DESCRIPTION (provided by applicant): The cerebral cortex controls mammalian behavior essentially through processing sensory input to then coordinate efferent activity via projection to subcortical structures. Anatomically, it is composed of a complex six-layer laminar structure. Sensory information processing occurs in the more superficial layers; cortical projection neurons (CPNs) arise in the deep layers and project axons either to other parts of the cortex or to subcortical areas, thus representing the most important cellular modulators of cortical output. Reestablishment of a CPN's complex, specific connectivity would therefore be an important goal of any repair strategy for cortical injuries. Recently, there has been much interest in using cellular therapy to replace neurons, usually through implantation of embryonic stem cells (ES cells). Although these cells can be readily induced to differentiate into neurons, it remains to be established whether there are qualitative differences between various differentiating strategies and types of neurons seen after transplantation. Our own studies assessing two pools of in vitro conditioned neuronal ES cells revealed an unexpected qualitative difference in their capacity to form CPN's after transplantation into neonatal cortex. Thus, while one cell pool produced virtually no CPN's, the other produced a robust subcortical projection that was remarkably anatomically appropriate. Furthermore, by serial comparisons of these two pools of cells, we were able to establish that this behavior was driven by a relatively small subset of transcription factors that function during development to produce the CPN population. These findings therefore raise the possibility that one can specifically modulate the neuronal phenotype of a cell repair treatment prior to transplantation through induction (or inhibition) of finite numbers of genes in vitro. In this R21 proposal, we propose to optimize the production of this specific neuronal cell subset through enriching the cell subpopulation for markers associated with CPN production. Additionally, we will assess the necessity and sufficiency of the identified genes in this process through either inducing their activity in the non- CPN producing pool of cells or silencing them in the CPN producing pool. With the results obtained, we will then be in a better position to assess whether this strategy has the capability of effecting functional improvement, thus moving it closer to eventual utility in the clinic, primarily as a treatment of cortical birth injuries. Brain injuries represent a significant public heath problem that is not only quality of life diminishing but also extremely costly. Improving recovery from these devastating problems is therefore of great relevance. Embryonic stem cells represent possibly the best potential cell type for use in brain transplantation. The ability to control the neuronal differentiation of these cells so that they can produce specific cell subsets should therefore be of great value in developing better therapeutic strategies for this problem.

描述（由申请人提供）：大脑皮层基本上通过处理感官输入来控制哺乳动物行为，然后通过投影对皮层结构来协调传出活动。从解剖学上讲，它由复杂的六层层状结构组成。感官信息处理发生在更浅层的层中。皮质投影神经元（CPN）在深层和轴突中出现到皮质的其他部位或皮质下区域，因此代表了皮质输出的最重要的细胞调节剂。因此，重建CPN复杂的特定连通性将是任何维修策略的重要目标。最近，通常是通过植入胚胎干细胞（ES细胞）来替代神经元的兴趣。尽管可以很容易地诱导这些细胞分化为神经元，但在移植后看到的各种区分策略和神经元的类型之间存在定性差异，还有待确定。我们自己的研究评估了两个体外调节神经元ES细胞的两个池，发现其在移植到新生儿皮层后形成CPN的能力存在意想不到的质量差异。因此，尽管一个细胞池几乎没有CPN，但另一个细胞池产生了强大的皮层投影，在解剖学上非常适合。此外，通过对这两个细胞池的序列比较，我们能够确定这种行为是由相对较小的转录因子的子集驱动的，这些转录因子在发育过程中起作用以产生CPN种群。因此，这些发现提出了一种可能性，即通过诱导（或抑制）在体外诱导（或抑制）移植前调节细胞修复治疗的神经元表型。在此R21提案中，我们建议通过丰富与CPN产生相关的标记的细胞亚群来优化该特定神经元细胞子集的产生。此外，我们将通过诱导其在非CPN产生细胞库中的活性或在CPN产生的池中沉降它们的活性来评估所鉴定基因的必要性和充分性。通过获得的结果，我们将有更好的位置来评估该策略是否具有影响功能改善的能力，从而使其更接近诊所的最终效用，主要是对皮质出生损伤的治疗。脑损伤代表了一个重大的公共卫生问题，不仅生活质量正在减少，而且昂贵。因此，改善这些毁灭性问题的恢复是很大的相关性。胚胎干细胞可能是用于脑移植的最佳潜在细胞类型。因此，控制这些细胞的神经元分化以使它们可以产生特定的细胞亚群的能力在为此问题开发更好的治疗策略方面具有很大的价值。