Cilia as a biomarker of CNS vascular health

纤毛作为中枢神经系统血管健康的生物标志物

• 批准号: 10252928
• 负责人: Surya Nauli
• 金额: $ 62.8万
• 依托单位: MEDICAL COLLEGE OF WISCONSIN
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-03 至 2022-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10252928
• 关键词: Active Biological Transport; Address; Adherence; Adhesions; Apical; Astrocytes; Biological Markers; Biology; Blood; Blood - brain barrier anatomy; Blood Vessels; Blood flow; Brain; Brain Diseases; Brain Injuries; Cell Adhesion; Cell Count; Cell surface; Cells; Cellular Morphology; Cellular biology; Cephalic; Cerebral Infarction; Cerebrovascular Circulation; Cerebrum; Cilia; Cilium Microtubule; Development; Disease; Endothelial Cells; Endothelium; Erythrocytes; Excision; Extravasation; Health; Hemorrhage; Human; Impairment; In Vitro; Injury; Knowledge; Length; Liquid substance; Mammals; Mediating; Microtubules; Mission; Modeling; Monitor; Mus; Neurons; Nude Mice; Organelles; Patients; Pericytes; Permeability; Phase; Phenotype; Preventive measure; Proteins; Protocols documentation; Public Health; Research; Research Personnel; Resistance; Rodent; Rodent Model; Role; Sickle Cell; Sickle Cell Anemia; Side; Signal Transduction; Structure; Surface; Testing; Transgenic Organisms; Traumatic Brain Injury; United States National Institutes of Health; Zebrafish; base; blood-brain barrier disruption; blood-brain barrier function; brain endothelial cell; cell type; clinically relevant; in vitro Model; in vivo; in vivo Model; induced pluripotent stem cell; innovation; model development; mouse model; neurovascular; neurovascular unit; recruit; sensor; severe injury

Project Summary The role of brain endothelial cells (ECs) and in particular the role of cilium, a microtubule flow sensor organelle expressed on the apical surface of ECs and facing the lumen has been poorly studied in the context of blood- brain-barrier (BBB) function. ECs and in turn cilia are the first line of contact with red blood cells (RBCs) in the blood. Studies from us and others suggest that cilia on ECs are critical for flow-mediated brain vessel stability. Thus, how flow relays signals to ECs via cilia and in turn to cells of the neurovascular (NVU) unit is not known. Without this knowledge, our ability to generate BBB models that mimic in vivo conditions will be hampered. The question of EC-cilia and its role in BBB function is clinically relevant given that patients with sickle cell disease (SCD) are predisposed to both overt and silent cerebral infarct, caused by sickle RBCs adhesion to the endothelium. The adhesion of sickle RBCs to endothelium may facilitate the physical removal of cilia (deciliation) from ECs surface, a mechanism recently identified in mammalian cilia shedding. In this multi-PI proposal, investigative team in vascular biology, sickle cell biology, human induced pluripotent stem cell (iPSC)-derived BBB and NVU models, cilia biology and rodent injury models will investigate the overarching hypothesis that disturbed cerebral blood flow triggers deciliation and release of cilia into the blood, thus resulting in aberrant signaling in the deciliated ECs and in the surrounding cells that comprise the NVU resulting in impact on NVU and BBB (Fig. 1). The objective of this proposal is to study RBCs-EC-cilia interaction and its importance to BBB model development. In the R61 phase, we will test whether RBCs from sickle cell disease (mouse models & human patients) will directly or indirectly trigger cilia shedding in brain ECs in vitro and in vivo. We will identify proteins in the cilia shed fragments, and also assess the importance of EC-cilia on BBB phenotypes in vitro and in vivo with emphasis on BBB integrity. Upon successfully establishing the milestone that RBC-EC-cilia interaction is critical for BBB function in the R61 phase, a go-decision in the R33 phase will initiate deeper probe into the underlying mechanisms and the role of the RBC-EC cilia interaction in SCD and traumatic brain injury (TBI) rodent models in vivo. The significance of this project is that EC-cilia status is an important determinant when brain ECs are included in flow-mediated BBB model development in vitro. The innovation is that until now, EC-cilia has been largely ignored in BBB protocols and accomplishing the objectives of this proposal will move the status quo in this field. This proposal will also bring us one step closer to monitoring cerebral vessel impairment using RBC-triggered EC-cilia shedding as a biomarker of vascular health, a NIH mission-related topic of research.

项目概要 脑内皮细胞 (EC) 的作用，特别是纤毛（一种微管流量传感器细胞器）的作用 在 EC 的顶端表面表达并面向管腔的表达在血液背景下的研究很少。 脑屏障（BBB）功能。 EC 和纤毛是与红细胞 (RBC) 接触的第一道线 血。我们和其他人的研究表明，内皮细胞上的纤毛对于血流介导的脑血管稳定性至关重要。 因此，流如何通过纤毛将信号传递给内皮细胞，进而传递给神经血管（NVU）单元的细胞尚不清楚。 如果没有这些知识，我们生成模拟体内条件的 BBB 模型的能力将会受到阻碍。这 鉴于镰状细胞病患者，EC-纤毛及其在 BBB 功能中的作用的问题具有临床相关性 (SCD) 容易发生明显的和无症状的脑梗塞，这是由镰状红细胞粘附到脑组织引起的。 内皮细胞。镰状红细胞与内皮的粘附可能有助于纤毛的物理去除 （脱纤毛）来自 EC 表面，这是最近在哺乳动物纤毛脱落中发现的一种机制。在这个多PI 提案，血管生物学、镰状细胞生物学、人类诱导多能干细胞研究小组 (iPSC) 衍生的 BBB 和 NVU 模型、纤毛生物学和啮齿动物损伤模型将研究总体情况 假设脑血流受到干扰会引发纤毛脱落并释放到血液中，因此 导致脱毛内皮细胞和构成 NVU 的周围细胞中信号传导异常 从而对NVU和BBB产生影响（图1）。本提案的目的是研究 RBCs-EC-cilia 相互作用及其对 BBB 模型开发的重要性。在R61阶段，我们将测试红细胞是否来自 镰状细胞病（小鼠模型和人类患者）将直接或间接触发大脑中的纤毛脱落 EC 体外和体内。我们将鉴定纤毛脱落片段中的蛋白质，并评估其重要性 EC-纤毛对体外和体内 BBB 表型的影响，重点是 BBB 完整性。成功建立后 RBC-EC-纤毛相互作用对于 R61 阶段的 BBB 功能至关重要的里程碑，这是 R33阶段将启动对RBC-EC纤毛的潜在机制和作用的更深入探讨 体内 SCD 和创伤性脑损伤 (TBI) 啮齿动物模型中的相互作用。该项目的意义在于 当脑 EC 包含在血流介导的 BBB 模型中时，EC 纤毛状态是一个重要的决定因素 体外发育。创新之处在于，到目前为止，EC-cilia 在 BBB 协议中基本上被忽略了 实现该提案的目标将改变该领域的现状。该提案还将带来 使用红细胞触发的 EC 纤毛脱落作为监测脑血管损伤的方法又近了一步 血管健康的生物标志物，NIH 任务相关的研究主题。

项目成果

sVEGFR1 Is Enriched in Hepatic Vein Blood-Evidence for a Provisional Hepatic Factor Candidate?

SVEGFR1富含肝静脉血液传播，成为临时肝脏因子候选者吗？

• DOI: 10.3389/fped.2021.679572
• 发表时间: 2021
• 期刊: Frontiers in pediatrics
• 影响因子: 2.6
• 作者: Spearman AD;Gupta A;Pan AY;Gudausky TM;Foerster SR;Konduri GG;Ramchandran R
• 通讯作者: Ramchandran R

Cilia proteins are biomarkers of altered flow in the vasculature.

• DOI: 10.1172/jci.insight.151813
• 发表时间: 2022-03-22
• 期刊: JCI insight
• 影响因子: 8
• 作者: Gupta A;Thirugnanam K;Thamilarasan M;Mohieldin AM;Zedan HT;Prabhudesai S;Griffin MR;Spearman AD;Pan A;Palecek SP;Yalcin HC;Nauli SM;Rarick KR;Zennadi R;Ramchandran R
• 通讯作者: Ramchandran R

Defined Differentiation of Human Pluripotent Stem Cells to Brain Microvascular Endothelial-Like Cells for Modeling the Blood-Brain Barrier.

• DOI: 10.1007/978-1-0716-3287-1_10
• 发表时间: 2023
• 期刊: Methods in molecular biology (Clifton, N.J.)

Established, New and Emerging Concepts in Brain Vascular Development.

• DOI: 10.3389/fphys.2021.636736
• 发表时间: 2021
• 期刊: Frontiers in physiology
• 影响因子: 4
• 作者: Gupta A;Rarick KR;Ramchandran R
• 通讯作者: Ramchandran R

Integrated Excitatory/Inhibitory Imbalance and Transcriptomic Analysis Reveals the Association between Dysregulated Synaptic Genes and Anesthetic-Induced Cognitive Dysfunction.

• DOI: 10.3390/cells11162497
• 发表时间: 2022-08-11
• 期刊: CELLS
• 影响因子: 6
• 作者: Yan, Yasheng;Logan, Sarah;Liu, Xiaojie;Chen, Bixuan;Jiang, Congshan;Arzua, Thiago;Ramchandran, Ramani;Liu, Qing-song;Bai, Xiaowen
• 通讯作者: Bai, Xiaowen