Designer Proteoglycans for Central Nervous System Injury

用于中枢神经系统损伤的设计蛋白多糖

• 批准号: 7647920
• 负责人: Diane M Snow
• 金额: $ 32.6万
• 依托单位: UNIVERSITY OF KENTUCKY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-08-01 至 2012-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7647920
• 关键词: Adult; Astrocytes; Behavior; Binding; Biological Assay; Brain; Chondroitin Sulfate Proteoglycan; Cicatrix; Core Protein; Cues; Data; Engineering; Family member; Goals; Growth; In Vitro; Injury; Knowledge; Lesion; Link; Measures; Modeling; Modification; Natural regeneration; Nerve; Nervous System Trauma; Nervous system structure; Neurites; Neuronal Injury; Neuronal Plasticity; Neurons; Patients; Pattern; Post-Translational Protein Processing; Protein Family; Protein Isoforms; Proteoglycan; Regulation; Relative (related person); Reporting; Site; Spinal Cord; Structure; Tertiary Protein Structure; Testing; Therapeutic; Up-Regulation; axon growth; central nervous system injury; clinical application; glycosylation; imaging modality; in vivo; injured; innovation; mutant; novel; progesterone 11-hemisuccinate-(2-iodohistamine); proteoglycan core protein; response; restoration; spinal cord and brain injury; sulfation

DESCRIPTION (provided by applicant): In the adult mammalian brain and spinal cord, neuronal injury results in failed regeneration, in part due to the upregulation of chondroitin sulfate proteoglycans (CSPGs). The majority of CSPGs originate from reactive astrocytes of the glial scar surrounding the lesion. The glial scar is beneficial for the recovering nervous system and should not be "excised", but axonal growth could be promoted if the expression of specific inhibitory portions of CSPGs could be targeted selectively. Our previous data indicate that neuronal inhibition is due to specific CSPG microheterogeneities and to the specific configurations of CSPGs predicted by their structure and by the molecules to which they bind. Our goal is to identify the most significant of the CSPG motifs with respect to neurite inhibition and regeneration, and specifically, to manipulate these moieties to promote regeneration. To this end, we and our collaborators have engineered 1) an array of CSPG isoforms and mutants we call "Designer PCs", 2) a variety of unique bioassays to express CSPGs, including a novel model of the glial scar in vitro, and 3) imaging methods to measure subtle features of nerve responses to CSPGs. The hypothesis of this proposal is: Identification and manipulation of specific inhibitory CSPG motifs using designer PGs and novel models of the glial scar will promote plasticity and regeneration in vitro and in vivo. We propose three Specific Aims that represent independent but interrelated studies to test this hypothesis. One, we will determine the expression and relative abundance of specific, inhibitory CSPG types and posttranslational modifications or core protein domains by reactive astrocytes in vitro. Two, we will determine the inhibitory potential for specific CSPG posttranslational modifications or core protein domains by establishing an inhibitory quotient to evaluate the responses of adult neurons in vitro. Three, we will manipulate specific inhibitory moieties of CSPGs to promote neuronal regeneration of adult neurons in vivo. The long term goal of this study is to identify the mechanism(s) of CSPG-induced inhibition following brain and spinal cord injury. The significance of the studies lies in the tremendous potential for translational application through the manipulation of specific CSPG motifs in injured patients, providing a therapeutic avenue to stimulate neuronal plasticity, facilitate reconnectivity of injured neurons, and accomplish restoration of function.

描述（由申请人提供）：在成年哺乳动物脑和脊髓中，神经元损伤导致再生失败，部分原因是硫酸软骨素蛋白聚糖（CSPGS）的上调。大多数CSPG源自病变周围的神经胶质疤痕的反应性星形胶质细胞。神经胶质疤痕对恢复神经系统是有益的，不应“切除”，但是如果可以选择性地靶向CSPG的特定抑制部分，则可以促进轴突生长。我们先前的数据表明，神经元抑制是由于特定的CSPG微分泌性以及由其结构和与其结合的分子预测的CSPG的特定构型引起的。我们的目标是确定有关神经突抑制和再生的最重要的CSPG基序，特别是操纵这些部分以促进再生。为此，我们和我们的合作者已经设计了1）我们称为“设计器PC”的CSPG同工型和突变体，2）表达CSPG的各种独特的生物测定，包括在体外的神经胶质疤痕模型，以及3）成像方法，以测量NEVER对CSPG的微妙特征。该提案的假设是：使用设计器PG和新型胶质疤痕模型对特定抑制性CSPG基序的识别和操纵将促进体外和体内的可塑性和再生。我们提出了三个代表独立但相互关联的研究以检验这一假设的特定目标。第一，我们将在体外通过反应性星形胶质细胞来确定特异性，抑制性CSPG类型和翻译后修饰或核心蛋白质结构域的表达和相对丰度。第二，我们将通过建立一种抑制商来评估体外的成年神经元反应，从而确定特定CSPG翻译后修饰或核心蛋白质结构域的抑制潜力。第三，我们将操纵CSPG的特定抑制部分，以促进体内成年神经元的神经元再生。这项研究的长期目标是确定脑和脊髓损伤后CSPG诱导的抑制作用的机制。研究的重要性在于通过操纵受伤患者的特定CSPG基序具有巨大的转化应用潜力，提供了刺激神经元可塑性的治疗途径，促进受伤的神经元的重新连接，并实现功能的恢复。