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

描述（由申请人提供）：该基金专注于转化研究，以推进神经成像技术，这将提高人类神经干细胞（hNSC）植入治疗新生儿缺氧缺血性脑损伤（HII）的潜力。我们的目标是在缺氧的单侧大脑中动脉闭塞的标准大鼠模型中使用先进的磁共振成像（MRI）（Rice- Vannucci，RVM）：（1）非侵入性监测（如临床上可能进行的）hNSC迁移、增殖、定位和作为结果生物标志物;（2）使用成像来定制最佳植入（部位、剂量和时间）;和（3）基于成像模型损伤严重性制定候选选择标准。使用11.7T MRI，我们开发了：（1）自动3D MRI体积方法来测量总的和局部的HII体积;（2）基于MRI的大鼠幼仔评分系统（RPSS）;（3）HII严重程度的3层模型（轻度、中度、重度）基于3D HI体积和RPSS;（4）3个可量化的神经影像学参数使用菲立磁标记的hNSC，可以与4种可定量的组织学，hNSC命运的免疫组织化学和神经生理学参数（整合、分化、连接和存活）。这些数据将在3个月时根据一系列广泛的行为测试进行评价。目的1a将检查神经成像参数是否与hNSC命运的参数相关。目的1b将检查铁标记是否对神经干细胞产生不利影响或导致额外的组织损伤。目的2将评估神经影像学的有效性，以确定植入部位、剂量和时间，以实现最佳结构和代谢恢复。目的3将使用神经影像学检查hNSC改善结构、代谢和行为结果的能力是否取决于使用RVM和双侧损伤的新模型（双侧颈动脉闭塞缺氧; BCAO-H）的初始HII的严重程度。该补助金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，损伤部位和损伤体积，我们已经开发了一种独特的和客观的方法来评估hNSC植入的疗效。hNSC治疗是治疗HII的一种非常有前途的方法，但我们有义务在开始新生儿临床试验之前以仔细、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：这项资助的重点是转化研究，以推进神经成像技术，这将提高人类神经干细胞（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（如可能在临床上完成）hNSC迁移，增殖和位置;（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人。使用11.7T MRI在两个模型缺血和缺氧，我们已经开发了自动化的三维MRI体积的方法来测量总的和区域HII体积和NSC迁移。这一提议将对用hNSC治疗新生儿HII产生重大影响。开发客观量化损伤、确认植入精度、监测hNSC活性和量化脑体积恢复的自动化MRI方法对于转化研究至关重要，因为这是在更高级物种中进行试验以及随后进行临床试验的前奏。

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.