Reciprocal communication between human neural stem cells and human endothelial cells

项目总结 神经干/祖细胞(NSPC)与血管形成的内皮细胞密切联系 (ECS)在发育中的大脑和成体神经干细胞的缝隙中，如脑室下区。此外, NSPC移植刺激卒中后新血管形成，促进功能恢复 在啮齿动物模型中。尽管NSPC和EC的互动很重要，但复杂的相互沟通 NSPC和ECs之间的关系还不是很清楚，特别是在人类身上。 血管内皮细胞对NSPC影响的研究主要集中在调节NSPC的EC分泌因子上 增殖、分化和自我更新。然而，EC接触对NSPC表型和NSPC的影响 人NSPC与内皮细胞的相互作用尚未得到很好的研究。我们证明了人类NSPC (HNSPC)与人内皮细胞(HECS)接触刺激表达两者的细胞百分比增加 GFAP和SOX2，这是B细胞的标志，B细胞是成人脑室下区的NSPC。第一个目标 这项研究的目的是确定hNSPC与HECS接触是否通过以下方式促进B细胞表型 联合培养中发现的GFAP/Sox2细胞的特征，通过对附加的B细胞标志物进行染色，单一 细胞RNA测序，细胞周期动力学和分化潜能的评估。第二个目标是 通过关注HEC接触介导的hNSPC表型改变，确定与此相关的机制 参与细胞间通讯的通路有N-钙粘素/b-连环蛋白、整合素信号、Notch信号、 和伊夫/伊夫林途径。NSPC刺激内皮细胞形成血管。然而，通过这些机制 NSPC刺激血管形成尚不清楚。使用3D神经血管模型的初步数据 证明了hNSPC分泌的因子促进了人类血管的形成。第三个目标是 通过评估hNSPC条件筛选刺激血管形成的hNSPC分泌成分 潜在的促血管生成可溶性因子和细胞外小泡的培养基及确认它们在 通过阻断它们的作用来形成血管。 利用人类细胞研究内皮细胞和神经前体细胞之间的相互作用可以更好地了解 它们的交流在人脑发育、成体干细胞生态位的调节和修复中的作用 脑部受伤。动物模型有助于研究的进步，但在翻译方面存在局限性 由于物种的差异。使用人类细胞将使我们能够研究两种细胞类型在 这个系统可能更接近于人脑生理，但在一个不那么复杂的环境中。在这下面 奖学金，我将有机会与加州大学欧文分校进行神经科学的领先研究人员合作， 在合作和支持的环境中进行干细胞和血管生物学研究。要扩展我的技术支持 为了获得技能和知识，我将参加研讨会、研讨会和会议，讨论对我的研究有重要意义的主题。 研究成果将通过会议和出版物与科学界和公众分享。

PROJECT SUMMARY Neural stem/progenitor cells (NSPCs) are in close communication with vessel-forming endothelial cells (ECs) in the developing brain and adult neural stem cell niches such as the subventricular zone. In addition, transplantation of NSPCs stimulates new vessel formation after stroke, leading to improved functional recovery in rodent models. Despite the importance of NSPC and EC interactions, the complex reciprocal communication between NSPCs and ECs is not well understood, especially in humans. Research on the effect of ECs on NSPCs has focused on EC-secreted factors, which regulate NSPC proliferation, differentiation, and self-renewal. However, the role of EC contact on NSPC phenotype and the interaction of human NSPCs and ECs have not been well studied. We demonstrated that human NSPC (hNSPC) contact with human ECs (hECs) stimulates an increase in the percentage of cells expressing both GFAP and Sox2, which are markers for type B cells, the NSPCs of the adult subventricular zone. The first aim of this study is to determine whether hNSPC contact with hECs promotes a type B cell phenotype by characterizing GFAP+/Sox2+ cells found in co-cultures via staining for additional type B cell markers, single cell RNA sequencing, assessment of cell cycle kinetics and differentiation potential. The second aim is to identify mechanisms involved in this hEC contact-mediated change in hNSPC phenotype by focusing on pathways involved in cell-cell communication such as N-cadherin/b-catenin, integrin signaling, Notch signaling, and Eph/ephrin pathways. NSPCs stimulate vessel formation by ECs. However, the mechanisms by which NSPCs stimulate vessel formation are not understood. Preliminary data using a 3D neurovascular model demonstrates that increased human vessel formation is promoted by hNSPC-secreted factors. The third aim is to identify hNSPC-secreted components stimulating hEC vessel formation by assessing hNSPC conditioned media for potential pro-vasculogenic soluble factors and extracellular vesicles and confirming their role in vessel formation by blocking their effects. Studying the interaction between ECs and NSPCs using human cells can provide better insight into the role of their communication in human brain development, regulation of adult stem cell niches, and repair after brain injury. Animal models have been instrumental for progress in research but have translational limitations due to species differences. Using human cells will allow us to study the interaction of both cell types in a system that may more closely resemble human brain physiology but in a less complex environment. Under this fellowship, I will have the opportunity to work with leading researchers at UC Irvine conducting neuroscience, stem cell, and vessel biology research in a collaborative and supportive environment. To expand my technical skills and knowledge, I will attend workshops, seminars, and conferences on topics important for my research. Research findings will be shared with the scientific community and public via conferences and publications.