Embryonic stem cell therapy after cervical contusion SCI in NHPs

项目概要/摘要 这是我们加州脊髓损伤（SCI）联盟提出的一项新的多PI提案， 从啮齿动物到灵长类动物的神经干细胞（NSC）移植的令人兴奋的结果，并评估 在我们的非人灵长类动物（NHP）颈挫伤SCI模型中的有效性和安全性。我们的多中心 联合会已经在一系列研究中检查了功能的恢复及其解剖学相关性， 颈椎半切模型。我们发现皮质脊髓束具有自发的广泛可塑性 (CST)这一系统在以前的啮齿动物研究中没有得到重视。我们开发了第一个大型NHP 颈椎半挫伤SCI模型以及开放场地评分系统和新型笼内前肢活动 和手部功能测试来评估功能结果。丰富的新信息和方向， 这种共享方法的价值，使用非常有价值的灵长类动物模型。该项目的重点是翻译 我们的NHP干细胞研究 我们现在报道，神经干细胞（NSC）来源于人脊髓移植早期半切部位， NHP SCI，在很长的距离上延伸非常大量的轴突，这些移植似乎 促进手部功能的长期恢复，并支持CST再生进入移植物。神经干细胞来源于 批准人胚胎干细胞H9（H9 hESCNSCs）也支持CST再生进入脊髓 脊髓损伤后NHP中的移植物。此外，我们已经推进了我们的细胞治疗策略，以产生第一个H9 hESCNSCs 尾向脊髓的命运，并已表明，这些细胞在啮齿动物移植， 促进CST轴突更有力的再生7.因此，在NHPs中，我们将移植 在更慢性和临床相关的六周时间点将hESCNSC尾化成挫伤损伤。我们 假设这些移植物将支持强健CST再生并增强前肢功能的恢复， 并为CST轴突提供中继以影响C8-T1脊髓中的前肢回路。我们将使用顺行 逆行追踪、免疫组化和移植细胞转染，并将连接数据与恢复相关联， 并测试这些移植物的长期存活率、安全性和功能效果。

PROJECT SUMMARY/ABSTRACT This is a new multi-PI proposal from our California Spinal Cord Injury (SCI) consortium to continue to translate exciting results from the transplantation of neural stem cells (NSCs) from rodent to primate, and to evaluate efficacy and safety in our non-human primate (NHP) cervical contusion SCI model. Our multi-center consortium has examined recovery of function and its anatomical correlates in a series of studies using a cervical hemisection model. We have discovered spontaneous and extensive plasticity of the corticospinal tract (CST) system that had not been appreciated in previous rodent studies. We have developed the first large NHP model of cervical hemicontusion SCI together with an open-field scoring system and novel in-cage forelimb activity and hand function tests to evaluate functional outcomes. The wealth of new information and directions speak to the value of this shared approach to using the very valuable primate model. This project focuses on translation of our NHP stem cell work. We now report that neural stem cells (NSCs) derived from human spinal cord grafted early to hemisection sites in NHP SCI, extend very large numbers of axons over very long distances, and that these transplants appear to enhance long-term recovery of hand function, and support CST regeneration into the graft. NSCs derived from the approved human embryonic stem cell H9 (H9 hESCNSCs) also support CST regeneration into spinal cord grafts in the NHP after SCI. Further, we have advanced our cell therapy strategy to produce the first H9 hESCNSCs caudalized to move them towards a spinal cord fate, and have shown that transplants of these cells in rodents promote much more vigorous regeneration of CST axons7. Therefore, in this proposal in NHPs, we will transplant caudalized hESCNSCs into a contusion lesion at a more chronic and clinically relevant six week time point. We hypothesize that these grafts will support robust CST regeneration and enhance recovery of forelimb function, and provide a relay for CST axons to influence forelimb circuitry in the C8-T1 cord. We will use anterograde and retrograde tracing, IHC and transfection of graft cells and correlate the connectional data with recovery, and test the long-term survival, safety, and functional effects of these transplants.