TRANSPLANTATION OF FETAL LGE IN HUNTINGTONS MODELS

摘要：描述(摘自申请者的摘要)：拟议研究的最终目标是为临床医生提供实施神经组织移植治疗亨廷顿病所必需的数据和技术。具体目标1将解决移植细胞是否可以改善与HD相关的认知缺陷。我们将在移植前和移植后的HD大鼠模型中使用延迟交替和空间学习记忆测试来评估这一点。这些措施将与移植物的生长、存活和移植物的神经化学表型相关，以确定对认知功能恢复重要的移植物特征。移植的有效性必须在进化上与人类关系更密切的动物模型中进行评估，以便测试更复杂的行为指标，并评估该程序的临床适用性。特定目标2在非人类灵长类动物HD模型中解决了这些问题。将细胞移植到3-硝基丙酸损毁的恒河猴尾状核和壳核中。猴子在移植前和移植后将通过录像带评估编舞运动、口面部运动障碍、肌张力障碍和手动灵活性的发生率进行行为测试。将进行进一步的测试，以评估认知影响，包括被动回避和视觉辨别任务。在行为测试结束时，这些动物将被宰杀，它们的大脑将被取出进行组织学处理。移植物的生长和存活及其细胞表型将与行为数据相关联，以评估移植物的疗效和临床应用的潜力。在临床上，细胞移植程序最重要的弱点之一是缺乏可靠的方法来监测移植物的发育。在具体目标3中，将使用1-H核磁共振波谱(NMRS)来监测患有兴奋性纹状体损伤的大鼠和猴子的组织移植，该技术利用MEGA(一种频率选择性重聚焦技术)来监测体内的GABA水平。在同步实验中，纹状体内损伤和移植物的大鼠将使用NMRS进行监测。使用9.4特斯拉磁铁在损伤前后对大鼠纹状体进行成像/测量。将定期进行体积、形态和免疫组织化学测量，并与从成像研究中获得的值和每只大鼠的行为测量相关联。成像数据将与实验结束时从灵长类受试者获得的最终组织学和行为测量相关联，以评估这种NRMS技术在组织移植中的临床有效性和适用性。

Abstract: DESCRIPTION (Adapted from applicant's abstract): The ultimate objective of the proposed research is to provide clinicians with data and techniques requisite for the implementation of transplantation of neural tissue in the treatment of Huntington's Disease (HD). Specific Aim 1 will address whether transplanted cells can ameliorate the cognitive deficits associated with HD. We will assess this using delayed alternation and spatial learning and memory tests pre- and post-transplantation in a rat model of HD. These measures will be correlated with graft growth, survival, and the neurochemical phenotype of the grafts to identify graft characteristics which are important for recovery of cognitive function. Transplant efficacy must be assessed in an animal model evolutionarily more closely related to humans in order to test more complex behavioral measures and to evaluate the clinical applicability of the procedure. Specific Aim 2 addresses these issues in a non-human primate model of HD. Cells will be transplanted into rhesus monkeys with 3-nitropropionic acid lesions of the caudate and putamen. Monkeys will be tested behaviorally pre- and post-transplantation by videotape assessment of incidence of choreiform movements, orofacial dyskinesia, dystonia, and manual dexterity. Further tests will be conducted to assess cognitive effects, including passive avoidance and visual discrimination tasks. At the end of the behavioral testing the animals will be sacrificed and their brains removed for histological processing. Growth and survival of the graft, as well as its cellular phenotype, will be correlated with behavioral data to evaluate graft efficacy and potential for clinical application. Clinically, one of the most important weaknesses of cell transplantation procedures is the absence of a reliable method for monitoring graft development. In Specific Aim 3, tissue grafts in rats and monkeys with excitotoxic striatal lesions will be monitored using 1-H nuclear magnetic resonance spectroscopy (NMRS) which utilizes MEGA, a frequency selective refocusing technique, to monitor GABA levels in vivo. In simultaneous experiments, rats with intrastriatal lesions and grafts will be monitored using NMRS. The striata of rats will be imaged/measured using the 9.4 Tesla magnet pre- and post-lesioning. Volumetric, morphologic and immunohistochemical measures will be taken periodically and correlated with the values obtained from the imaging studies and with behavioral measures from each rat.. The imaging data will be correlated with the final histological and behavioral measures obtained from the primate subjects at the end of the experiments to evaluate the clinical efficacy and applicability of this NRMS technique in tissue transplantation.