Reciprocal communication between human neural stem cells and human endothelial cells

人类神经干细胞和人类内皮细胞之间的相互通讯

• 批准号: 10391313
• 负责人: Brenda Gutierrez
• 金额: $ 4.24万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-12-01 至 2023-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10391313
• 关键词: 3-Dimensional; Adopted; Adult; Affect; Animal Model; Astrocytes; B cell differentiation; B-Lymphocytes; Biology; Blood Vessels; Brain; Brain Injuries; Cadherins; Cell Communication; Cell Cycle Kinetics; Cell Therapy; Cells; Coculture Techniques; Communication; Communities; Complex; Data; Development; Educational workshop; Endothelial Cells; Environment; Ephrins; Fellowship; Future; Glial Fibrillary Acidic Protein; Goals; Human; In Vitro; Injury; Integrins; Knowledge; Ligands; Mass Spectrum Analysis; Mediating; Mediator of activation protein; Membrane Proteins; Modeling; N-Cadherin; Neurosciences; Pathway interactions; Phenotype; Physiology; Production; Property; Proteins; Publications; Recovery of Function; Regulation; Research; Research Personnel; Rodent; Rodent Model; Role; Signal Transduction; Stains; System; Technical Expertise; Testing; Therapeutic; Transplantation; Work; adult stem cell; beta catenin; brain cell; cell behavior; cell type; collaborative environment; exosome; extracellular vesicles; human adult stem cell; human disease; immunocytochemistry; improved; insight; intercellular communication; nerve stem cell; neurovascular; notch protein; novel; post stroke; prevent; receptor; relating to nervous system; repaired; scaffold; self-renewal; single-cell RNA sequencing; species difference; stem; stem cell proliferation; stem cells; subventricular zone; supportive environment; symposium; transcriptome; transcriptome sequencing

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.

项目摘要 神经茎/祖细胞（NSPC）与血管形成的内皮细胞密切相关 （EC）在发育中的大脑和成人神经干细胞壁细胞（例如脑室下区）中。此外， NSPC的移植刺激中风后新容器的形成，从而改善了功能恢复 在啮齿动物模型中。尽管NSPC和EC相互作用很重要，但复杂的相互交流 在NSPC和EC之间，尤其是在人类之间。 对EC对NSPC的影响的研究集中在调节NSPC的EC分泌因素上 扩散，差异化和自我更新。但是，EC接触在NSPC表型和 人类NSPC和EC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EC（HEC）接触可刺激表达两者的细胞百分比的增加 GFAP和SOX2，它们是B型细胞的标记，是成年室内下区的NSPC。第一个目标 这项研究的是确定HNSPC是否与HEC的接触是否促进了B型细胞表型 表征在共培养中发现的GFAP+/SOX2+细胞通过染色的其他类型B细胞标记，单个 细胞RNA测序，细胞周期动力学评估和分化潜力。第二个目标是 通过专注于此HEC接触介导的HEC介导的变化的机制，通过着重于HNSPC表型 参与细胞 - 细胞通信的途径，例如N-钙粘蛋白/B-catenin，整联蛋白信号传导，Notch信号传导， 和EPH/Ephrin途径。 NSPC通过EC刺激血管形成。但是， 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 证明人血管形成增加的是HNSPC分泌因子促进。第三个目标是 通过评估有条件的HNSPC，识别刺激HEC血管形成的HNSPC分泌的组件 培养基，用于潜在的促血管生成因子和细胞外囊泡，并确认其在 通过阻断其作用来形成船只。 使用人类细胞研究EC和NSPC之间的相互作用可以更好地洞悉 他们在人类脑发育中的沟通作用，成年干细胞壁ches的调节以及修复之后 脑损伤。动物模型对研究的进展有用，但具有翻译局限性 由于物种差异。使用人类细胞将使我们能够研究两种细胞类型在A中的相互作用 系统可能更像人类脑生理，但在不太复杂的环境中。在此 奖学金，我将有机会与UC Irvine的领先研究人员合作，进行神经科学， 干细胞和船只生物学研究在协作和支持性的环境中。扩大我的技术 技能和知识，我将参加对我的研究重要的主题研讨会，研讨会和会议。 研究发现将通过会议和出版物与科学界和公众共享。