Investigating Pericyte Roles in Blood-Brain Barrier Formation

研究周细胞在血脑屏障形成中的作用

• 批准号: 10390466
• 负责人: ERIC V SHUSTA
• 金额: $ 36.78万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-05-01 至 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10390466
• 关键词: Adult; Alzheimer' s Disease; Astrocytes; BMP5 gene; Biological Assay; Blood; Blood - brain barrier anatomy; Blood Circulation; Blood Vessels; Brain; Brain Diseases; Cell Therapy; Cells; Characteristics; Data; Demyelinating Diseases; Development; Disease; Disease model; Embryo; Embryonic Development; Endothelial Cells; Endothelium; Epilepsy; Functional disorder; Gene Expression Profile; Generations; Genetic; Genomic approach; Human; Immune; In Vitro; Invaded; Knowledge; Laboratories; Leukocyte Adhesion Molecules; Maintenance; Mesoderm; Modeling; Molecular; Multiple Sclerosis; Mus; Neural Crest; Neuraxis; Neurons; Pathology; Pathway interactions; Patients; Pericytes; Peripheral; Permeability; Phenotype; Plasma Proteins; Play; Process; Property; Regulation; Reporting; Role; Signal Transduction; Signaling Molecule; Somatic Cell; Stroke; Structure; Supplementation; Supporting Cell; System; Testing; Therapeutic; Tight Junctions; Transport Process; Traumatic Brain Injury; Tube; base; blood-brain barrier function; brain endothelial cell; cell type; cohort; developmental disease; fetal; genome editing; human model; improved; in vitro Model; induced pluripotent stem cell; induced pluripotent stem cell technology; innovation; insight; mouse model; nervous system disorder; neurovascular unit; novel; progenitor; programs; receptor; restoration; stem cell differentiation; tool; transcriptome; transcriptomics; transcytosis

ABSTRACT The blood-brain barrier (BBB) acts as a signaling and transport interface between the blood and brain. The BBB begins to form early in embryonic development as the mesoderm-derived vasculature invades the immature central nervous system (CNS) and acquires BBB characteristics such as tight junctions and a lack of fenestrae. After further maturation, the adult BBB, with its very low permeability and a wealth of molecular transport systems, is maintained by interactions with supporting cells of the neurovascular unit (NVU). Recent studies have indicated the importance of CNS pericytes in BBB formation, with pericytes triggering reduced transcytosis, reduced expression of leukocyte adhesion molecules and proper tight junction organization. However, the identity of pericyte-derived factors that can elicit these important changes during BBB formation are not known. Thus, our understanding of the molecular mechanisms underpinning BBB formation is incomplete; and in this proposal, we aim to further examine the mechanisms by which brain pericytes impact BBB formation. A powerful and innovative approach to explore BBB formation is the use of human induced pluripotent stem cell (iPSC) technology to model the BBB and the associated support cells of the NVU. We will demonstrate that not only can brain pericytes be differentiated from iPSCs, they can also regulate key BBB properties in iPSC-derived brain endothelial cells (BMECs). In parallel, using genomics approaches, we have identified a cohort of pericyte-derived secreted factors, several of which can induce BBB properties in iPSC- derived BMECs. Combining these approaches, we will examine the mechanisms whereby pericyte-derived secreted factors can differentially regulate BBB formation in iPSC-derived BMECs. Preliminary data indicate that one of the pericyte-derived secreted factors, BMP5, can influence hallmark BBB properties known to be regulated by brain pericytes. Namely, BMP5 can reduce transcytosis and improve tight junction structures in iPSC-derived BMECs. To further examine the mechanism by which BMP5 regulates BBB formation during development, we will use genetic mouse models to explore whether BMP5 signaling is necessary for BBB formation and function. Finally, we will assess whether BMP5 supplementation can be therapeutic in a mouse model of multiple sclerosis. Understanding the pericyte-derived regulators of BBB formation could yield many new mechanistic insights regarding brain diseases that have demonstrable pericyte involvement. Knowledge of the barrier formation pathways could also open new avenues for restoring BBB function in debilitating neurological disease.

抽象的 血脑屏障（BBB）充当血液和大脑之间的信号传递和转运界面。这 随着中胚层的脉管系统入侵，BBB在胚胎发育中开始形成 未成熟的中枢神经系统（CNS），并获得BBB特征，例如紧密连接和缺乏 Fenestrae。进一步成熟后，成年BBB的渗透性非常低，大量分子 通过与神经血管单元（NVU）的支撑细胞相互作用来维持运输系统。最近的 研究表明，CNS周细胞在BBB形成中的重要性，周细胞触发降低 跨胞菌病，白细胞粘附分子的表达降低和适当的紧密连接组织。 但是，周细胞衍生的因素的身份可以在BBB形成期间引起这些重要变化 不知道。因此，我们对BBB形成的分子机制的理解是 不完整；在此提案中，我们旨在进一步研究脑周细胞影响的机制 BBB组。一种强大而创新的探索BBB形成的方法是使用人类诱导的 多能干细胞（IPSC）技术，以建模NVU的BBB和相关的支持细胞。我们将 证明不仅可以将脑周细胞与IPSC区分开，还可以调节关键BBB IPSC衍生的脑内皮细胞（BMEC）中的特性。同时，使用基因组学方法，我们有 确定了一系列周细胞衍生的分泌因素，其中一些可以诱导IPSC-的BBB特性 派生的BMEC。结合了这些方法，我们将研究周围衍生的机制 分泌的因素可以差异地调节IPSC衍生的BMEC中的BBB形成。初步数据表示 BMP5的衍生的周细胞分泌因素之一可以影响已知的Hallmark BBB特性 受脑周细胞的调节。也就是说，BMP5可以减少跨胞菌病并改善紧密连接结构 IPSC衍生的BMEC。进一步检查BMP5在调节BBB形成期间的机制 开发，我们将使用遗传小鼠模型来探索BMP5信号是否需要BBB 形成和功能。最后，我们将评估补充BMP5是否可以在小鼠中进行治疗 多发性硬化症的模型。了解BBB形成的周细胞衍生的调节剂可能会产生许多 关于具有可证明的有阴极参与的脑疾病的新机械见解。知识 障碍构成途径还可以为恢复BBB功能的新途径开放 神经疾病。

项目成果

Intrinsic blood-brain barrier dysfunction contributes to multiple sclerosis pathogenesis.

内在的血脑屏障功能障碍导致多发性硬化症的发病机制。

• DOI: 10.1093/brain/awac019
• 发表时间: 2022
• 期刊: Brain : a journal of neurology
• 作者: Nishihara,Hideaki;Perriot,Sylvain;Gastfriend,BenjaminD;Steinfort,Marel;Cibien,Celine;Soldati,Sasha;Matsuo,Kinya;Guimbal,Sarah;Mathias,Amandine;Palecek,SeanP;Shusta,EricV;Pasquier,RenaudDu;Engelhardt,Britta
• 通讯作者: Engelhardt,Britta

Differentiation of human pluripotent stem cells to brain microvascular endothelial cell-like cells suitable to study immune cell interactions.

• DOI: 10.1016/j.xpro.2021.100563
• 发表时间: 2021-06-18
• 期刊: STAR protocols
• 作者: Nishihara H;Gastfriend BD;Kasap P;Palecek SP;Shusta EV;Engelhardt B
• 通讯作者: Engelhardt B