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.

项目摘要 脑内皮细胞（EC）的作用，尤其是微管传感器细胞器Cilium的作用 在血液的背景下，在EC的顶部表面和面向管腔的表面表达很差 脑磁力磁带（BBB）功能。 ECS和CILIA是与红细胞（RBC）的第一线接触 血。我们和其他人的研究表明，EC的纤毛对于流动介导的脑血管稳定性至关重要。 因此，流动如何通过纤毛传递到EC和ECS并又转移到神经血管（NVU）单位的细胞中。 没有这些知识，我们产生模仿体内疾病的BBB模型的能力将受到阻碍。这 鉴于患有镰状细胞病的患者，EC-CILIA的问题及其在BBB功能中的作用与临床相关 （SCD）易受镰状RBC粘附引起的明显和沉默的脑梗塞 内皮。镰状RBC对内皮的粘附可能有助于物理去除纤毛 （分解）来自ECS表面，该机制最近在哺乳动物的纤毛脱落中鉴定出来。在这个多pi中 提案，血管生物学，镰状细胞生物学，人类诱导多能干细胞的调查小组 （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 扰乱脑血流的假设会触发纤毛的分解并释放到血液中，因此 导致构成NVU的分解EC和周围细胞中的异常信号传导 导致对NVU和BBB的影响（图1）。该建议的目的是研究RBCS-EC-Cilia 互动及其对BBB模型开发的重要性。在R61阶段，我们将测试RBC是否来自 镰状细胞疾病（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在体外和体内。我们将确定纤毛碎片中的蛋白质，并评估 EC-CILIA在体外和体内对BBB表型，重点是BBB完整性。成功建立 RBC-EC-CILIA相互作用的里程碑对于R61阶段的BBB函数至关重要 R33阶段将对基本机制和RBC-EC纤毛的作用进行更深入的探测 SCD和创伤性脑损伤（TBI）啮齿动物模型的相互作用。该项目的重要性是 当流量介导的BBB模型中包括脑ECS时，EC-CILIA状态是重要的决定因素 体外发展。创新是，到目前为止，EC-Cilia在BBB协议中很大程度上被忽略了 实现此提案的目标将移动该领域的现状。该提议也将带来 我们接近使用RBC触发的EC-CILIA脱落来监测大脑容器损伤的一步 血管健康的生物标志物，这是NIH与任务有关的研究主题。

项目成果

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

• 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