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 神经干/祖细胞（NSPCs）与血管形成内皮细胞（Vessel-forming endothelial cells，VES）有着密切的联系 (ECs)在发育中的大脑和成人神经干细胞龛，如脑室下区。此外，本发明还提供了一种方法， 移植NSPCs可刺激卒中后新血管形成，从而改善功能恢复 在啮齿动物模型中。尽管NSPC和EC相互作用的重要性，复杂的相互沟通， NSPCs和EC之间的关系尚不清楚，特别是在人类中。 关于内皮细胞对神经干细胞作用的研究主要集中在内皮细胞分泌的调节神经干细胞的因子上 增殖、分化和自我更新。然而，EC接触对NSPC表型的作用和 人NSPCs和ECs的相互作用尚未得到很好的研究。我们证明了人类NSPC 与人内皮细胞（hEC）接触刺激表达两者的细胞百分比增加。 GFAP和Sox 2，它们是B型细胞的标志物，即成年脑室下区的NSPCs。第一个目标 本研究的目的是确定hNSPC与hEC接触是否通过以下方式促进B型细胞表型： 通过对另外的B型细胞标记物进行染色来表征在共培养物中发现的GFAP+/Sox 2+细胞， 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 通过关注以下方面，确定hNSPC表型中hEC接触介导的变化所涉及的机制 参与细胞-细胞通讯的途径如N-钙粘蛋白/β-连环蛋白，整合素信号传导，Notch信号传导， 和Eph/ephrin途径。NSPCs通过EC刺激血管形成。然而， NSPC刺激血管形成的机制尚不清楚。使用3D神经血管模型的初步数据 证明增加的人血管形成是由hNSPC分泌的因子促进的。第三个目标是 通过评估hNSPC条件化来鉴定刺激hEC血管形成的hNSPC分泌组分 潜在的促血管生成可溶性因子和细胞外囊泡的培养基，并证实它们在 通过阻断其作用形成血管。 使用人类细胞研究EC和NSPCs之间的相互作用可以更好地了解 它们的通讯在人类大脑发育中的作用，成体干细胞龛的调节，以及 脑损伤动物模型有助于研究的进展，但具有转化的局限性 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的相互作用， 可能更接近于人脑生理学但在较不复杂环境中的系统。根据本 奖学金，我将有机会与加州大学欧文分校的主要研究人员进行神经科学， 干细胞和血管生物学研究在一个合作和支持的环境。为了扩大我的技术 技能和知识，我将参加研讨会，研讨会和会议的主题对我的研究很重要。 研究成果将通过会议和出版物与科学界和公众分享。