Hippocampal regulation and neuroprotection by NPC-derived VEGF

描述（由申请人提供）：干细胞疗法在治疗人类疾病，特别是目前无法治愈的神经退行性疾病方面具有很大的前景。然而，为了有效地利用干细胞的愈合潜力，有必要了解这些细胞如何与它们的环境相互作用并作出反应。在移植中，干细胞可以作为替代细胞，对局部线索作出反应，帮助修复受损组织。它们也可能作为保护性信号的来源，在未分化状态下分泌促进局部修复过程的生长因子。这一建议侧重于干细胞作为神经调节分泌细胞的第二种，不太为人所知的作用。本研究的目的是研究成人神经干/祖细胞（NPC）来源的分泌物在海马中的调节和神经保护作用。考虑到海马体在一些神经退行性疾病如颞叶癫痫和阿尔茨海默病中的显著退化，这一大脑区域为干细胞治疗提供了一个有吸引力的靶点。Wyss-Coray实验室先前的研究表明，分离的成年海马npc大量分泌多种生长因子，最显著的是具有高度神经保护作用的血管内皮生长因子（VEGF）。我的目标是研究NPC来源的VEGF调节海马功能并提供退行性疾病的神经保护的假设。在目标1中，我将确定npc衍生的VEGF如何在体外和体内使用npc特异性的基因敲除技术调节其他npc的增殖和分化。在Aim 2中，我将使用Aim 1中的体内敲低模型来确定NPC来源的VEGF如何调节成年小鼠的局部血管环境和海马行为。最后，在目标3中，我将在神经损伤的兴奋毒性模型中测试来自内源性NPC（和NPC移植）的NPC衍生的VEGF的神经保护特性。这些研究将有助于提供对npc如何在体内作为未来疗法发挥作用的关键理解。这个项目将基础神经科学与转化研究相结合，需要我掌握广泛的技能

DESCRIPTION (provided by applicant): Stem cell therapies hold great promise for treatment of human disease, particularly for currently incurable neurodegenerative diseases. However, to effectively harness the healing potential of stem cells, it is necessary to understand how these cells interact with and respond to their environment. In transplants, stem cells may function as replacement cells that respond to local cues to help mend damaged tissue. They may also function as a source of protective cues, secreting in their undifferentiated state growth factors that facilitate local repair processes. This proposal focuses on this second, less well understood role of stem cells as neuromodulatory secretory cells. The goal of this proposal is to investigate the regulatory and neuroprotective role of adult neural stem/progenitor cell (NPC)-derived secretions in the hippocampus. Given the prominent degradation of the hippocampus in several neurodegenerative disorders such as temporal lobe epilepsy and Alzheimer's disease, this brain area provides an attractive target for stem cell therapy. The Wyss-Coray lab has previously shown that isolated adult hippocampal NPCs secrete a variety of growth factors in large quantities, most notably the highly neuroprotective vascular endothelial growth factor (VEGF). My goal is to investigate the hypothesis that NPC- derived VEGF regulates hippocampal function and provides neuroprotection from degenerative disease. In Aim 1, I will determine how NPC-derived VEGF regulates the proliferation and differentiation of other NPCs in vitro and in vivo using genetic knockdown techniques specific to NPCs. In Aim 2, I will use the in vivo knockdown models from Aim 1 to determine how NPC- derived VEGF regulates the local vascular environment and hippocampal behavior in adult mice. Finally, in Aim 3, I will test the neuroprotective qualities of NPC-derived VEGF from endogenous NPCs (and from NPC transplants) in an excitotoxic model of neural insult. These studies will help provide critical understanding of how NPCs may function as future therapies in vivo. This project combines basic neuroscience with translational research, requiring that I become proficient in a broad skill set. I will gain experience in in vitro NPC culture, shRNA, lox- cre genetics, stereotaxic injectios into the hippocampus and neurodegenerative models of disease. Through this project, it is my goal to acquire expertise in translational research designed to pursue treatment for neurodegenerative disease.