Neonatal Brain Ischemia: Neuroimaging as a Basis For Rational Stem Cell Therapy

描述（由申请人提供）：该资助重点用于推进神经成像技术的转化研究，这些技术将增强人类神经干细胞（hNSC）植入治疗新生儿缺氧缺血性脑损伤（HII）的潜力。我们的目标是在单侧大脑中动脉闭塞伴缺氧（Rice- Vannucci， RVM）的标准幼鼠模型中使用先进的磁共振成像（MRI）来：(1)无创监测（如临床上可能做的）hNSC迁移、增殖、位置并作为结果生物标志物；(2)利用成像技术定制最佳植入（部位、剂量和时间）；(3)根据影像学模型损伤严重程度制定候选选择标准。使用11.7T MRI，我们开发了：(1)一种自动3D MRI体积测量方法来测量总和区域HII体积；(2)基于MRI的大鼠幼仔评分系统（RPSS）；(3)基于三维HI体积和RPSS的HII严重程度3层模型（轻度、中度、重度）；(4)使用feridex标记hNSC的3个可量化神经影像学参数（迁移、增殖和最终定位），可与hNSC命运的4个可量化组织学、免疫组织化学和神经生理学参数（整合、分化、连通性和生存）进行比较。这些数据将在3个月后通过一系列广泛的行为测试进行评估。目的1a将检查神经影像学参数是否与hNSC的预后相关。Aim 1b将检查铁标记是否对NSCs产生不利影响或引起额外的组织损伤。目的2将评估神经影像学的有效性，以确定最佳结构和代谢恢复的植入部位、剂量和时间。目的3将使用神经影像学检查hNSCs改善结构、代谢和行为结果的能力是否取决于初始HII的严重程度，使用RVM和新的双侧损伤模型（双侧颈动脉闭塞伴缺氧；BCAO-H）。这项拨款解决了许多必须考虑的重要技术和生物学问题，以提高NSC治疗成功的机会。通过使用先进的成像方法来确定HII的严重程度、损伤位置和损伤体积，我们开发了一种独特而客观的方法来评估hNSC植入的疗效。hNSC治疗是一种非常有前途的治疗HII的方法，但在对新生儿进行临床试验之前，我们有义务以谨慎、客观、合乎逻辑和安全的方式提供科学基础。公共卫生相关性：该资助侧重于推进神经成像技术的转化研究，这些技术将增强人类神经干细胞（hNSC）植入治疗新生儿缺氧缺血性脑损伤（HII）的潜力。我们的目标是使用先进的磁共振成像（MRI）技术作为结果测量，并：(1)监测非侵入性（如临床上可能做的）hNSC迁移，增殖和定位；(2)优化hNSC植入部位、剂量和时机；(3)根据损伤严重程度的早期MRI测量确定潜在的治疗候选者。在缺血和缺氧两种模型中使用11.7T MRI，我们开发了自动3D MRI体积方法来测量总和区域HII体积和NSC迁移。这一建议将对用hNSCs治疗新生儿HII产生重大影响。开发能够客观量化损伤、确认植入精度、监测hNSC活动和量化脑容量恢复的自动化MRI方法对于转化研究至关重要，这是高阶物种试验和临床试验的前导。

DESCRIPTION (provided by applicant): This grant focuses on translational research to advance neuroimaging techniques that will enhance the potential of human neural stem cell (hNSC) implantation to treat neonatal hypoxic- ischemic brain injury (HII). Our goal is to use advanced magnetic resonance imaging (MRI) in a standard rat pup model of unilateral middle cerebral artery occlusion with hypoxia (Rice- Vannucci, RVM) to: (1) monitor non-invasively (as might be done clinically) hNSC migration, proliferation, location and as an outcome biomarker; (2) use imaging to tailor optimal implantation (site, dose, and timing); and (3) develop candidate selection criteria based on imaging model injury severity. Using an 11.7T MRI, we have developed: (1) a automated 3D MRI volumetric method to measure total and regional HII volumes; (2) a MRI based Rat Pup Scoring System (RPSS); (3) a 3-tiered model of HII severity (mild, moderate, severe) based on 3D HI volumes and the RPSS; and (4) 3 quantifiable neuroimaging parameters (migration, proliferation and final location) using Feridex-labeled hNSCs that can be compared with 4 quantifiable histological, immunohistochemistry and neurophysiological parameters of hNSC fate (integration, differentiation, connectivity, and survival). These data will be evaluated at 3 months against an extensive battery of behavioral testing. Aim 1a will examine whether neuroimaging parameters correlate with parameters of hNSC fate. Aim 1b will examine if iron labeling adversely affects NSCs or causes additional tissue injury. Aim 2 will assess the efficacy of neuroimaging to determine site, dose, and timing of implantation for optimal structural and metabolic recovery. Aim 3 will use neuroimaging to examine whether the ability of hNSCs to improve structural, metabolic and behavioral outcomes depends upon the severity of the initial HII using the RVM and a new model of bilateral injury (bilateral carotid occlusion with hypoxia; BCAO-H). This grant addresses many of the important technical and biological issues that must be considered in order to improve the chance for success of NSC therapy. By using advanced imaging methods to determine HII severity, injury location and injury volumes, we have developed a unique and objective approach to evaluate efficacy of hNSC implantation. hNSC treatment is an extremely promising approach to treat HII, but we have an obligation to provide the scientific foundation in a careful, objective, logical and safe manner before embarking on clinical trials in newborns. PUBLIC HEALTH RELEVANCE: This grant focuses on translational research to advance neuroimaging techniques that will enhance the potential of human neural stem cell (hNSC) implantation to treat neonatal hypoxic- ischemic brain injury (HII). Our goals are to use advanced magnetic resonance imaging (MRI) techniques as an outcome measure and to: (1) monitor non-invasively (as might be done clinically) hNSC migration, proliferation and location; (2) optimize hNSC implantation site, dose, and timing; and (3) identify potential therapeutic candidates based on early MRI measures of injury severity. Using an 11.7T MRI in two models ischemia and hypoxia we have developed automated 3D MRI volumetric methods to measure total and regional HII volumes and NSC migration. This proposal will have a significant impact on treating neonatal HII with hNSCs. Development of automated MRI methods that objectively quantify injury, confirm precision of implantation, monitor hNSC activity, and quantify brain volumetric recovery is critical for translational research as a prelude to trials in higher order species and then clinical trials.