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

描述(由申请者提供)：这笔赠款专注于提高神经成像技术的转化性研究，以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)在标准的一侧大脑中动脉阻塞伴缺氧(莱斯-万努奇，RVM)大鼠模型中：(1)无创性地(如临床上可能做的)监测hNSC的迁移、增殖、位置和作为结果生物标记物；(2)使用成像来定制最佳植入(位置、剂量和时机)；以及(3)根据成像模型的损伤严重程度制定候选选择标准。使用11.7T磁共振成像，我们已经开发出：(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PUP评分系统(RPSS)；(3)基于3D HI体积和RPSS的HII严重程度(轻、中、重度)的三层模型；以及(4)使用Feridex标记的hNSCs的3个可量化的神经成像参数(迁移、增殖和最终位置)，可以与hNSC命运的4个可量化的组织学、免疫组织化学和神经生理参数(整合、分化、连接和生存)相比较。这些数据将在3个月后根据广泛的行为测试进行评估。目的1a将检查神经影像参数是否与hNSC命运参数相关。目标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将评估神经成像的有效性，以确定植入的位置、剂量和时机，以实现最佳的结构和代谢恢复。目的3将使用神经成像检查hNSCs改善结构、代谢和行为结果的能力是否取决于使用RVM的初始HII的严重程度和双侧损伤的新模型(双侧颈动脉闭塞伴低氧；BCAO-H)。这笔赠款解决了许多必须考虑的重要技术和生物问题，以提高NSC治疗的成功机会。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：这笔赠款专注于促进神经成像技术的转化研究，该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)技术作为结果测量和：(1)非侵入性(如临床可能做的)监测hNSC的迁移、增殖和位置；(2)优化hNSC的植入位置、剂量和时机；以及(3)基于早期MRI损伤严重程度的测量来确定潜在的候选治疗方案。在脑缺血和缺氧两种模型中使用11.7T磁共振成像，我们发展了自动化的3D MRI体积测量方法来测量总的和局部的HII体积以及NSC的迁移。这项建议将对hNSCs治疗新生儿HII产生重大影响。发展能够客观量化损伤、确认植入的精确度、监测hNSC活性和量化脑容量恢复的自动化MRI方法对于翻译研究至关重要，它是更高级物种试验的前奏，然后是临床试验。

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.