Engineering In Vitro ECM Test Beds to Mimic Traumatic Neural Injury

模拟创伤性神经损伤的体外 ECM 试验台工程

基本信息

批准号：
9204863
负责人：
CHRISTINE E SCHMIDT
金额：
$ 21.89万
依托单位：
UNIVERSITY OF FLORIDA
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-02-01 至 2019-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9204863
关键词：
Animal Model Animals Antibodies Beds Biocompatible Materials Biological Models CSPG3 gene Cell physiology Central Nervous System Diseases Characteristics Chondroitin Sulfate Proteoglycan Cicatrix Cleaved cell Clinical Degenerative polyarthritis Disease Disease Progression Engineering Environment Epilepsy Extracellular Matrix Gel Gelatinase A Goals Hyaluronan In Vitro Injury Intervention Knowledge Matrix Metalloproteinases Mediating Modeling Neuraxis Neuroglia Neurons Outcomes Research Parkinson Disease Pathologic Pathology Patients Peptide Hydrolases Phenotype Play Proteoglycan Rattus Recovery Reproducibility Research Rodent Role Site Spinal Cord Spinal cord injury System Techniques Testing Therapeutic Time Tissue Engineering Tissues United States Work aggrecan base brevican cell behavior cost cost effective design experimental study in vitro testing in vivo injured janusin link protein nerve injury new therapeutic target novel novel therapeutics public health relevance response scaffold screening small molecule therapeutics versican

项目摘要

DESCRIPTION: The goal of this application is to develop an in vitro test bed capable of mimicking native and pathological extracellular matrix (ECM) to identify novel targets for treatment of traumatic neural injury. We will accomplish this goal through rational design of an ECM-based scaffold to mimic native temporal ECM damage observed after spinal cord injury (SCI). Chondroitin sulfate proteoglycans (CSPGs) are the largest component of the healthy and pathologic ECM in the central nervous system (CNS) and serve many functions. After SCI specifically, there is an increase in versican, neurocan, and brevican, and a decrease in aggrecan. There are also dynamic changes to specific proteases such at MMP-2 and -9 after SCI that will selectively cleave CSPGs. Furthermore, CSPG fragmentation has been implicated in progression of other diseases (e.g. osteoarthritis). We believe CSPG fragmentation plays a similar role in increasing unwanted glial scarring after SCI or traumatic neural injury. Currently, animal models are used to screen feasibility of biomaterials for tissue engineering. However, to examine injury and disease states an "induced" state must be created in the animal model that often does not represent native pathology of injury or disease. Creation of in vitro pathological ECM test beds has the potential to provide a lower cost, more relevant system for examining mechanistic responses and testing small molecule therapeutics and identifying relevant targets to treat patients with SCI. We hypothesize that neoepitopes exposed after specific proteoglycan fragmentation exacerbate progression of glial scarring after spinal cord injury, and an in vitro pathological ECM test bed is a novel platform to delineate the influence of these fragmentation profiles on glial cell phenotype. In Aim 1 we will identify the type and degree of aggrecan fragmentation at distinct time points after spinal cord injury (SCI) using tissue from previously executed studies in a rat model. In Aim 2 we will create 3D ECM from hyaluronan (hyaluronic acid, HA), HA-link protein 1, tenascin-R, and aggrecan (intact and fragmented) to mimic the ECM of the CNS after SCI. The final task will be to assess the temporal response of glial cells to these engineered gels by analyzing changes in their phenotype and function to assess their progression towards a reactive state. This research will enable the creation of in vitro test beds capable of isolating the role of fragmentation in the CNS after injury to help identify novel targes for clinical therapeutics. Furthermore, it will enhance our understanding of the interplay between CSPG fragmentation and pathological cell behavior after SCI. In conjunction with "body-on-a-chip" approaches, these biomaterials platforms hold the potential to revolutionize current screening techniques and ultimately eliminate animal screening completely. Application of these test beds could be broadened for use in diseases of the CNS and other tissues, where fragmentation of CSPGs has also been identified as a key player in disease progression (e.g., epilepsy, Parkinson's) to help isolate targets and identify novel therapies.

描述：本申请的目标是开发一种能够模拟天然和病理性细胞外基质 (ECM) 的体外测试床，以确定治疗创伤性神经损伤的新靶标。我们将通过合理设计基于 ECM 的模型来实现这一目标。模拟脊髓损伤（SCI）后观察到的天然颞叶 ECM 损伤的支架是中枢神经系统健康和病理 ECM 的最大组成部分。 (CNS) 并发挥多种功能，特别是在 SCI 后，多功能蛋白聚糖 (versican)、神经蛋白聚糖 (neurocan) 和短蛋白聚糖 (brevican) 增加，聚集蛋白聚糖 (aggrecan) 减少。SCI 后，MMP-2 和 -9 等特定蛋白酶也会发生动态变化。此外，CSPG 碎片与其他疾病（例如骨关节炎）的进展有关，我们认为 CSPG 碎片在增加不必要的疾病方面也发挥着类似的作用。目前，SCI 或创伤性神经损伤后的神经胶质疤痕。动物模型用于筛选用于组织工程的生物材料的可行性，但是，为了检查损伤和疾病状态，必须在动物模型中创建通常不代表体外病理学创建的“诱导”状态。 ECM 测试床有潜力提供成本更低、更相关的系统，用于检查机械反应和测试小分子疗法，并确定治疗 SCI 患者的相关靶标。脊髓损伤后的疤痕，体外病理 ECM 测试床是描述这些碎片特征对神经胶质细胞表型的影响的新平台。在目标 1 中，我们将确定脊髓损伤后不同时间点聚集蛋白聚糖碎片的类型和程度。使用先前在大鼠模型中进行的研究的组织进行脊髓损伤 (SCI) 在目标 2 中，我们将从透明质酸（透明质酸，HA）、HA 连接蛋白创建 3D ECM。 1、tenascin-R 和聚集蛋白聚糖（完整和片段化）来模拟 SCI 后 CNS 的 ECM 最终任务是通过分析神经胶质细胞表型和功能的变化来评估神经胶质细胞对这些工程凝胶的时间反应。他们向反应状态的进展这项研究将能够创建体外试验床。能够分离损伤后中枢神经系统中的碎片的作用，以帮助确定临床治疗的新靶标。此外，结合“body-on-a”，它将增强我们对 CSPG 碎片与病理细胞行为之间相互作用的理解。通过“芯片”方法，这些生物材料平台有可能彻底改变当前的筛选技术，并最终完全消除动物筛选，这些测试床的应用可以扩大到中枢神经系统和其他组织的疾病，其中 CSPG 会破碎。也被认为是疾病进展（例如癫痫、帕金森病）的关键因素，有助于分离靶标并确定新疗法。