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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.

产品说明：本申请的目的是开发一种能够模拟天然和病理性细胞外基质（ECM）的体外试验床，以鉴定用于治疗创伤性神经损伤的新靶点。我们将通过合理设计基于ECM的支架来模拟脊髓损伤（SCI）后观察到的原生颞ECM损伤来实现这一目标。硫酸软骨素蛋白聚糖（CSPG）是中枢神经系统（CNS）中健康和病理性ECM的最大组分，具有多种功能。SCI后，多功能蛋白聚糖、神经蛋白聚糖和短蛋白聚糖增加，聚集蛋白聚糖减少。在SCI后，特异性蛋白酶如MMP-2和MMP-9也有动态变化，其将选择性地切割CSPG。此外，CSPG片段化与其他疾病（例如骨关节炎）的进展有关。我们相信CSPG断裂在SCI或创伤性神经损伤后增加不必要的胶质疤痕方面发挥着类似的作用。目前，动物模型用于筛选用于组织工程的生物材料的可行性。然而，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损伤或疾病的天然病理学的“诱导”状态。体外病理ECM测试床的创建有可能提供更低成本、更相关的系统，用于检查机制反应和测试小分子疗法并鉴定治疗SCI患者的相关靶点。我们假设特异性蛋白聚糖片段化后暴露的新表位加剧了脊髓损伤后神经胶质瘢痕形成的进展，体外病理性ECM试验床是一种新的平台，可以描述这些片段化特征对神经胶质细胞表型的影响。在目标1中，我们将使用来自先前在大鼠模型中执行的研究的组织，在脊髓损伤（SCI）后的不同时间点鉴定聚集蛋白聚糖片段化的类型和程度。在目标2中，我们将从透明质酸（透明质酸，HA），HA连接蛋白1，生腱蛋白-R和聚集蛋白聚糖（完整和片段）中创建3D ECM，以模拟SCI后CNS的ECM。最后的任务将是通过分析其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响应，以评估其向反应状态的进展。这项研究将使体外试验床的创建成为可能能够分离损伤后CNS中碎片化的作用，以帮助鉴定临床治疗的新靶点。此外，它将增强我们对SCI后CSPG片段化和病理细胞行为之间相互作用的理解。结合“芯片上的身体”方法，这些生物材料平台有可能彻底改变目前的筛选技术，并最终完全消除动物筛选。这些测试床的应用可以扩大到用于CNS和其他组织的疾病，其中CSPG的片段化也被确定为疾病进展的关键因素（例如，癫痫，帕金森氏症），以帮助隔离目标和确定新的疗法。